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Inhibition of bladder cancer cell proliferation by allyl isothiocyanate (mustard essential oil)
... A dosage of 15-30 µM BITC or PEITC was necessary to induce damage of UMUC3 bladder cancer cells [15], whereas T24 cell proliferation was suppressed already at 2-5 µM PEITC [16]. AITC at concentrations of >60 µM has been noted to block growth and proliferation of RT4 and T24 cells [17,18]. UMUC3 cells, however, responded to a lower dosage of 15 µM AITC in terms of cell growth inhibition [19]. ...
... The apoptotic machinery has not been further evaluated in the present investigation. Based on Sávio and coworkers, the mechanism of action of BITC, AITC, and PEITC on apoptotic pathways may differ among several bladder cancer cell lines [17]. Ongoing experiments should, therefore, deal with the fine-tuned action of these ITCs on the apoptotic cascade. ...
... AITC treatment reduced cell growth of both RT4 and T24 cells, albeit by a different mechanism. Cell death by apoptosis was the prominent underlying mechanism in RT4 cells, whereas blocking of cell cycle regulating proteins was the major action of AITC in T24 cells [17]. ...
Combined cisplatin–gemcitabine treatment causes rapid resistance development in patients with advanced urothelial carcinoma. The present study investigated the potential of the natural isothiocyanates (ITCs) allyl-isothiocyanate (AITC), butyl-isothiocyanate (BITC), and phenylethyl-isothiocyanate (PEITC) to suppress growth and proliferation of gemcitabine- and cisplatin-resistant bladder cancer cells lines. Sensitive and gemcitabine- and cisplatin-resistant RT112, T24, and TCCSUP cells were treated with the ITCs, and tumor cell growth, proliferation, and clone formation were evaluated. Apoptosis induction and cell cycle progression were investigated as well. The molecular mode of action was investigated by evaluating cell cycle-regulating proteins (cyclin-dependent kinases (CDKs) and cyclins A and B) and the mechanistic target of the rapamycin (mTOR)-AKT signaling pathway. The ITCs significantly inhibited growth, proliferation and clone formation of all tumor cell lines (sensitive and resistant). Cells were arrested in the G2/M phase, independent of the type of resistance. Alterations of both the CDK–cyclin axis and the Akt–mTOR signaling pathway were observed in AITC-treated T24 cells with minor effects on apoptosis induction. In contrast, AITC de-activated Akt–mTOR signaling and induced apoptosis in RT112 cells, with only minor effects on CDK expression. It is concluded that AITC, BITC, and PEITC exert tumor-suppressive properties on cisplatin- and gemcitabine-resistant bladder cancer cells, whereby the molecular action may differ among the cell lines. The integration of these ITCs into the gemcitabine-/cisplatin-based treatment regimen might optimize bladder cancer therapy.
... Their activation-induced effect is presumably based on Ca2+-mediated mechanisms, which could be addressed in future research. However, our results are in agreement with other findings obtained from HeLa cervical cancer cell line [49,50] and gastric cancer cells [51]. Furthermore, 24 h AITC treatment significantly reduced the viability of OSCC line at 100 nM and completely diminished the viability by 99% at 5 µM concentration. ...
... Liu et al. reported that a high dose (≥20 µM) of AITC decreased cell viability, increased DNA damage and inhibited cell migration in HepG2 human hepatocellular carcinoma cells [52]; moreover, AITC has antiangiogenic affect in in vivo studies [53]. A number of studies also showed that TRPA1 agonist AITC negatively affects the cell proliferation of breast [54], bladder [50] ...
... Their activation-induced effect is presumably based on Ca 2+ -mediated mechanisms, which could be addressed in future research. However, our results are in agreement with other findings obtained from HeLa cervical cancer cell line [49,50] and gastric cancer cells [51]. Furthermore, 24 h AITC treatment significantly reduced the viability of OSCC line at 100 nM and completely diminished the viability by 99% at 5 µM concentration. ...
Oral squamous cell carcinoma (OSCC) is a common cancer with poor prognosis. Transient Receptor Potential Ankyrin 1 (TRPA1) and Vanilloid 1 (TRPV1) receptors are non-selective cation channels expressed on primary sensory neurons and epithelial and immune cells. TRPV1 mRNA and immunopositivity, as well as TRPA1-like immunoreactivity upregulation, were demonstrated in OSCC, but selectivity problems with the antibodies still raise questions and their functional relevance is unclear. Therefore, here, we investigated TRPA1 and TRPV1 expressions in OSCC and analyzed their functions. TRPA1 and TRPV1 mRNA were determined by RNAscope in situ hybridization and qPCR. Radioactive 45Ca2+ uptake and ATP-based luminescence indicating cell viability were measured in PE/CA-PJ41 cells in response to the TRPA1 agonist allyl-isothiocyanate (AITC) and TRPV1 agonist capsaicin to determine receptor function. Both TRPA1 and TRPV1 mRNA are expressed in the squamous epithelium of the human oral mucosa and in PE/CA-PJ41 cells, and their expressions are significantly upregulated in OSCC compared to healthy mucosa. TRPA1 and TRPV1 activation (100 µM AITC, 100 nM capsaicin) induced 45Ca2+-influx into PE/CA-PJ41 cells. Both AITC (10 nM–5 µM) and capsaicin (100 nM–45 µM) reduced cell viability, reaching significant decrease at 100 nM AITC and 45 µM capsaicin. We provide the first evidence for the presence of non-neuronal TRPA1 receptor in the OSCC and confirm the expression of TRPV1 channel. These channels are functionally active and might regulate cancer cell viability.
... It has fungicidal and bactericidal activities against a plethora of microorganisms, and its anticancer activities were demonstrated in different cancer cell lines in vitro [10] and some animal tumor models in vivo [11]. Quite a few studies demonstrated that AITC elicits cell apoptosis and causes cell cycle arrest through modulation of various genes involved in proliferation and survival in malignant cell lines such as cervical cancer [10], colorectal carcinoma [9,12,13], breast adenocarcinoma [14], hepatoma [15], prostate cancer [11], bladder malignancy [16][17][18], brain glioma [19], and oral cancer [20]. ...
... This pattern of the treated cells undertaken apoptosis first and then turned into necrosis was also observed for the cells treated with AITC only. Our flow cytometric results showed similar trends to previously reported studies of AITC on different cancer models, i.e., the cell death was increased in a time-dependent manner [18,27,32,33]. ...
... This pattern of the treated cells undertaken apoptosis first and then turned into necrosis was also observed for the cells treated with AITC only. Our flow cytometric results showed similar trends to previously reported studies of AITC on different cancer models, i.e., the cell death was increased in a time-dependent manner [18,27,32,33]. The Caspase-3/7 green detection reagent has DEVD (four amino acid) peptide conjugated with green dye (excitation/emission: 503/530 nm) with the ability to bind with nucleic acid, but the dye is nonfluorescent until it cleaves from the DEVD peptide and binds to DNA. ...
Sinigrin is present in significant amounts in cruciferous vegetables. Epidemiological studies suggest that the consumption of such vegetables decreases the risk of cancer, and the effect is attributed mainly to allyl isothiocyanate (AITC), a hydrolysis product of sinigrin catalyzed by myrosinase. Anticancer activity of AITC has been previously investigated for several cancer models, but less attention was paid to delivering AITC on the target site. In this study, the gene sequences of core streptavidin (coreSA) and myrosinase (MYR) were cloned in a pET-30a(+) plasmid and transformed into BL21(DE3) E. coli competent cells. The MYR-coreSA chimeric protein was expressed and purified using immobilized metal affinity chromatography and further characterized by gel electrophoresis, Western blot, and enzyme activity assay. The purified MYR-coreSA chimeric protein was tethered on the outer membrane of biotinylated adenocarcinoma A549 cells and then treated with various concentrations of sinigrin. Our results showed that 20 µM of sinigrin inhibited the growth of A549 cells tethered with myrosinase by ~60% in 48 h. Furthermore, the levels of treated cells undertaken apoptosis were determined by Caspase-3/7 activation and Annexin-V. In summary, sinigrin harnessed like a prodrug catalyzed by myrosinase to the production of AITC, which induced cell apoptosis and arrested the growth of lung cancer cells.
... Combined treatment using AITC and sulforaphane (a glucosinolate-derived ITC) was found to synergistically multi-target the system of proliferation and metastasis in human non-small cell lung cancer A549 cells (18). AITC suppressed cell proliferation in bladder cancer RT4 and T24 cells via different mechanisms upon p53 gene expression (19). AITC was also found to exhibit antitumor effects against colorectal adenocarcinoma SW620 cells by inducing G2/M phase arrest and downregulating Cdc25B and Cdc25C levels (20). ...
... Moreover, blocking Ca 2+ -based ER stress signals and ROS formation significantly attenuated AITC-induced HT-29 cell apoptosis. The effective cytotoxic actions of AITC on multiple human cancer cell lines are well documented (12,(16)(17)(18)(19)(20)(21); however, the detailed mechanisms involved in the cytotoxic effects of AITC in HT-29 cells remain elusive. Our results indicated that AITC inhibited HT-29 cell growth and proliferation. ...
... AITC was found to facilitate type I programmed cell death (apoptosis) (16)(17)(18)(19)(20)(21) or induce autophagy (21) in various types of cancer cell lines. The present study investigated the apoptotic machinery in AITC-treated HT-29 cells. ...
Allyl isothiocyanate (AITC), a bioactive phytochemical compound that is a constituent of dietary cruciferous vegetables, possesses promising chemopreventive and anticancer effects. However, reports of AITC exerting antitumor effects on apoptosis induction of colorectal cancer (CRC) cells in vitro are not well elucidated. The present study focused on the functional mechanism of the endoplasmic reticulum (ER) stress‑based apoptotic machinery induced by AITC in human colorectal cancer HT‑29 cells. Our results indicated that AITC decreased cell growth and number, reduced viability, and facilitated morphological changes of apoptotic cell death. DNA analysis by flow cytometry showed G2/M phase arrest, and alterations in the modulated protein levels caused by AITC were detected via western blot analysis. AITC also triggered vital intrinsic apoptotic factors (caspase‑9/caspase‑3 activity), disrupted mitochondrial membrane potential, and stimulated mitochondrial‑related apoptotic molecules (e.g., cytochrome c, apoptotic protease activating factor 1, apoptosis‑inducing factor, and endonuclease G). Additionally, AITC prompted induced cytosolic Ca2+ release and Ca2+‑dependent ER stress‑related signals, such as calpain 1, activating transcription factor 6α, glucose‑regulated proteins 78 and 94, growth arrest‑ and DNA damage‑inducible protein 153 (GADD153), and caspase‑4. The level of reactive oxygen species (ROS) production was found to induce the hallmark of ER stress GADD153, proapoptotic marker caspase‑3, and calpain activity after AITC treatment. Our findings showed for the first time that AITC induced G2/M phase arrest and apoptotic death via ROS‑based ER stress and the intrinsic pathway (mitochondrial‑dependent) in HT‑29 cells. Overall, AITC may exert an epigenetic effect and is a potential bioactive compound for CRC treatment.
... The anti-cancer effects of AITC and its molecular mechanisms have been investigated in various BC cell lines; for instance, AITC was found to lead to morphological changes and inhibit the cell proliferation of the human BC cell lines RT4 and T24 [46]. AITC was also reported to affect cell cycle arrest and apoptosis of RT4 and T24 cells [44]. ...
... Moreover, the anti-cancer effects of AITC involved different molecular mechanisms between RT4 and T24 cells. In RT4 cells, AITC treatment increased S100P and Bax levels and decreased Bcl-2 levels; meanwhile, Bax, Bcl-2, and anillin levels increased while S100P levels decreased in T24 cells [46]. The Bax/Bcl-2 pathway is speculated to be a key modulator of AITC in RT4 cells, with anillin and S100P mainly functioning in this system [46]. ...
... In RT4 cells, AITC treatment increased S100P and Bax levels and decreased Bcl-2 levels; meanwhile, Bax, Bcl-2, and anillin levels increased while S100P levels decreased in T24 cells [46]. The Bax/Bcl-2 pathway is speculated to be a key modulator of AITC in RT4 cells, with anillin and S100P mainly functioning in this system [46]. Thus, these in vitro studies demonstrated that the anti-cancer effects of AITC in BC cells are dependent on the pathological and molecular characteristics of cancer cells. ...
Bladder cancer (BC) is a representative of urological cancer with a high recurrence and metastasis potential. Currently, cisplatin-based chemotherapy and immune checkpoint inhibitors are used as standard therapy in patients with advanced/metastatic BC. However, these therapies often show severe adverse events, and prolongation of survival is unsatisfactory. Therefore, a treatment strategy using natural compounds is of great interest. In this review, we focused on the anti-cancer effects of isothiocyanates (ITCs) derived from cruciferous vegetables, which are widely cultivated and consumed in many regions worldwide. Specifically, we discuss the anti-cancer effects of four ITC compounds—allyl isothiocyanate, benzyl isothiocyanate, sulforaphane, and phenethyl isothiocyanate—in BC; the molecular mechanisms underlying their anti-cancer effects; current trends and future direction of ITC-based treatment strategies; and the carcinogenic potential of ITCs. We also discuss the advantages and limitations of each ITC in BC treatment, furthering the consideration of ITCs in treatment strategies and for improving the prognosis of patients with BC.
... Tis is because the seed contains many polyphenolic compounds, including vitamins A, C, and E, calcium, and iron [1,2]. Additionally, they provide an important source of dietary antioxidants and have high radical scavenging activity in preventing various chronic diseases such as cancer and cardiovascular disease [3,4]. Based on the Central Bureau of Statistics [5], mustard production in Indonesia will reach approximately 667 thousand tons in 2020. ...
... Tis is a cultivation technique using water or nutrient solution as a growing medium, known as soilless culture [9]. Te main advantages are [10]as follows: (1) possible use of areas unsuitable for conventional agriculture, such as dry and degraded land; (2) plant independence for weather conditions; (3) year-round cultivation; and (4) reducing the use of labor-intensive activities such as weeding and soil preparation. Te activity in hydroponics is also arguably lighter than conventional farming [11,12]. ...
Agricultural land has been converted into settlements following the population growth in various parts of the country. The productivity of horticulture, particularly pagoda mustard (Brassica narinosa L), decreases with the narrowing of fields. The main milestone as a promising solution to overcoming this issue is applying the hydroponic technique. This study aims to analyze the potential of hydroponic kit-based growing on a self-fertigation system for pagoda mustard production. In contrast to general hydroponic, the proposed hydroponic kit is supported by a smart valve component as a unique novelty used for the automatic distribution of nutrients without electrical power (zero energy). The mustard seeds were sown on rockwool for two to three days in a dark room and placed in the sun for seventeen days. A total of 50 pagoda mustard seeds were arranged evenly on a self-fertigation system tray following a zig-zag planting pattern for forty days. The seed has the following morphological characteristics: average height of 22.88 cm, biomass width of 26.42 cm, root length of 23.4 cm, and weight of 241.5 g. Furthermore, the production requires a total fertigation consumption of 186 L (equal to 0.0935 L/plant day−1) with an actual crop coefficient between 0.01 and 0.54. The proposed system shows good performance for mustard growth with a uniformity value between 80 and 89%. Finally, hydroponic kit-based growing on a self-fertigation system can be applied in various areas to produce and maintain a sustainable food supply.
... Allyl isothiocyanate (AITC), one of the constituents of naturally occurring ITCs, and thus can also be found in cruciferous vegetables, exerts multiple biological effects, such as anti-microbial [13,14], anti-inflammatory [15,16], anti-angiogenic [17], and anti-cancer activities. It inhibits the proliferation of human bladder cancer cells [18] and suppresses the cell metastasis of colorectal adenocarcinoma [19], bladder cancer [20], and hepatoma [21] cells. It also induce apoptotic cell death in bladder cancer cells [18], cervical cancer cells [22], hepatoma cells [20], colorectal cancer cells [23], and human cisplatin-resistant oral cancer cells [24]. ...
... It inhibits the proliferation of human bladder cancer cells [18] and suppresses the cell metastasis of colorectal adenocarcinoma [19], bladder cancer [20], and hepatoma [21] cells. It also induce apoptotic cell death in bladder cancer cells [18], cervical cancer cells [22], hepatoma cells [20], colorectal cancer cells [23], and human cisplatin-resistant oral cancer cells [24]. Moreover, in in vivo studies, AITC has been shown to reduce liver fibrosis by regulating Kupffer cell activation [25], and to facilitate lipid accumulation and inflammation via the Sirt1/AMPK and NF-κB signaling pathways in rats with non-alcoholic fatty liver disease [16]. ...
Some clinically used anti-cancer drugs are obtained from natural products. Allyl isothiocyanate (AITC), a plant-derived compound abundant in cruciferous vegetables, has been shown to possess an anti-cancer ability in human cancer cell lines in vitro, including human brain glioma cells. However, the anti-cancer effects of AITC in human glioblastoma (GBM) cells in vivo have not yet been examined. In the present study, we used GBM8401/luc2 human glioblastoma cells and a GBM8401/luc2-cell-bearing animal model to identify the treatment efficacy of AITC. Here, we confirm that AITC reduced total cell viability and induced cell apoptosis in GBM8401/luc2 cells in vitro. Furthermore, Western blotting also showed that AITC induced apoptotic cell death through decreased the anti-apoptotic protein BCL-2, MCL-1 expression, increased the pro-apoptotic protein BAX expression, and promoted the activities of caspase-3, -8, and -9. Therefore, we further investigated the anti-tumor effects of AITC on human GBM8401/luc2 cell xenograft mice. The human glioblastoma GBM8401/luc2 cancer cells were subcutaneously injected into the right flank of BALB/c nude mice to generate glioblastoma xenograft mice. The animals were randomly divided into three groups: group I was treated without AITC (control); group II with 0.1 mg/day of AITC; and group III with 0.2 mg/day of AITC every 3 days for 27 days. Bodyweight, and tumor volume (size) were recorded every 3 days. Tumors exhibiting Luc2 intensity were measured, and we quantified intensity using Living Image software on days 0, 12, and 24. After treatment, tumor weight from each mouse was recorded. Tumor tissues were examined for histopathological changes using H&E staining, and we analyzed the protein levels via immunohistochemical analysis. Our results indicate that AITC significantly inhibited tumor growth at both doses of AITC due to the reduction in tumor size and weight. H&E histopathology analysis of heart, liver, spleen, and kidney samples revealed that AITC did not significantly induce toxicity. Body weight did not show significant changes in any experiment group. AITC significantly downregulated the protein expression levels of MCL-1, XIAP, MMP-9, and VEGF; however, it increased apoptosis-associated proteins, such as cleaved caspase-3, -8, and -9, in the tumor tissues compared with the control group. Based on these observations, AITC exhibits potent anti-cancer activity in the human glioblastoma cell xenograft model via inhibiting tumor cell proliferation and the induction of cell apoptosis. AITC may be a potential anti-GBM cancer drug that could be used in the future.
... In order to reduce these effects, the use of natural compounds has been studied in the last few years (Reis and Jones, 2017). Several natural compounds, such as allyl isothiocyanate (extracted from mustard oil), resveratrol (extracted from grape seeds), epigallocatechins (extracted from green tea), and garlic extracts have demonstrated antitumor effects in bladder carcinoma (Savio et al., 2015;Matsuo et al., 2017;Kim et al., 2018). ...
... To evaluate cell density after silibinin treatment, 2 × 10 4 T24, 5637, or RT4 cells were seeded into 12-well plates. After 24 hr, cells were treated with the silibinin solution at concentrations of 5, 10, and 0.4% trypan blue (Savio et al., 2015). Untreated cells were used as a negative control. ...
Silibinin, a natural compound extracted from milk thistle, has demonstrated antitumor properties in urinary bladder cancer cells; however the role of TP53 gene in these effects is unclear. In order to better understand the molecular and antiproliferative mechanisms of this compound, urinary bladder cancer cells with different TP53 gene status, RT4 (low grade tumor, wild TP53 gene), 5,637 (high grade tumor, grade 2, mutated TP53 gene) and T24 (high grade tumor, grade 3, mutated TP53 gene), were treated with several concentrations of silibinin (1; 5; 10; 50; 100 and 150 μM). Cytotoxicity, pro-oxidant effect, morphological changes, cell migration, cell cycle progression, global methylation profile and relative expression of HOXB3, c-MYC, PLK1, SMAD4, SRC, HAT, HDAC and RASSF1A genes were evaluated. The silibinin presented cytotoxic and pro-oxidant effects in the three cell lines. In mutated TP53 cells, significant interference in cell migration and cell cycle arrest at the G2/M phase was observed. Additionally, silibinin induced global DNA hypomethylation in the highest grade tumor cells. For wild-type TP53 cells, a sub-G1 apoptotic population was present. Furthermore, there was modulation of gene expression responsible for cell growth (SMAD and c-MYC), migration (SRC), cell cycle kinetics (PLK1), angiogenesis (HOXB3) and of genes associated with epigenetic events such as DNA acetylation (HAT) and deacetylation (HDAC). In conclusion, the silibinin inhibited the urinary bladder tumor cell proliferation independently of TP53 status; however cell cycle effects, gene expression changes and alteration of cell migration are dependent on TP53 status. This article is protected by copyright. All rights reserved.
... For instance, AITC may arrest cancer cells in mitosis, which in turn leads to apoptosis via B-cell lymphoma 2 (Bcl-2) protein phosphorylation (4)(5)(6). AITC may affect the proliferation of various tumor cells by inducing apoptosis and cell cycle arrest, or by inhibiting their invasion and metastasis, including in breast (7), bladder (8)(9)(10), lung (11), colon (12,13), liver (14,15) and prostate cancer cells (16). On the basis of the inhibitory effect AITC has on the proliferation of bladder cancer cells, population-based survey results have suggested that the intake of cruciferous plants may improve the survival rate of patients with bladder cancer (17), which may provide further evidence for the anticancer effect of AITC. ...
... The results of the present study suggested that the apoptosis of HeLa cells induced by AITC may be associated with an imbalance in Bcl-2/Bax expression. Sávio et al (8) also revealed that the apoptosis induced by AITC was associated with a Bcl-2/Bax expression imbalance. ...
The present study aimed to investigate the effect of allyl isothiocyanate (AITC) on the viability and apoptosis of the human cervical cancer HeLa cell line in vitro, and to explore the potential underlying mechanisms of this. HeLa cells were treated with varying concentrations of AITC for different durations. The cell viability was then measured using a Cell Counting kit-8 assay and the apoptosis rate of the cells was detected using flow cytometry. Additionally, the B cell lymphoma-2 (Bcl-2) and Bcl-2-associated X protein (Bax) mRNA expression levels were determined by reverse transcription-quantitative polymerase chain reaction, while the Bax and Bcl-2 protein expression levels in cells were detected by western blot analysis. AITC was revealed to inhibit the viability of HeLa cells. AITC was revealed to induce the apoptosis of HeLa cells, as the apoptosis rate increased gradually with an increase in the dose. As the concentration of AITC increased, the Bax mRNA expression level increased, whilst the Bcl-2 mRNA expression level decreased. Furthermore, the Bax protein expression intensity increased whilst Bcl-2 protein expression intensity decreased, thereby resulting in a decrease in the ratio of Bcl-2/Bax proteins. AITC may inhibit cell viability by inducing the apoptosis of HeLa cells and this may be accounted for by the imbalance in the Bcl-2/Bax expression ratio.
... These effects were observed as a result of cyclin B1, VEGF and Casp-3 inhibition (Bhattacharya et al. 2010b). Also, in wild type as well as mutated bladder cancer cell lines, allyl isothiocyanates could affect the gene expression of factors involved in cell apoptosis including S100P, BAX, and BCL2 (Savio et al. 2015). Another species of mustard, B. compestris, was assessed in an animal model of benzo(a)pyrene [B(a)P]-induced forestomach tumorigenesis and 3-methylcholantrene (MCA)-induced uterine cervix tumorigenesis. ...
... As cancer is a complicated disease with high mortality rate, it is conceivable that clinical investigation of anticancer drugs cannot be easily performed. Thus, more (Huang et al. 2000), induction of apoptosis through Casp activation, increased Bax expression, NF-ĸB down regulation and induction of PARP cleavage (Khan et al. 2016), increase in p53 gene specific mRNA (Yazdanpanahi et al. 2014), decrease invasion and metastasis via downregulation of COX-2, MMP-9, CXCR4, and VEGF (Park, 2011a, b), suppression of VEGFR2 signaling pathways and angiogenesis (Pang et al. 2009), induction of differentiation and inhibition of topoisomerase II (Zhao et al. 2003), inhibition of Akt phosphorylation, GSKb, and cyclin D1 (Chou et al. 2017) Human leukemia HL-60, K 562, U937, MOLT-4, THP-1 cell culture (Huang et al. 2000;Hostanska et al. 2002), human brain tumor (Hostanska et al. 2002), human colorectal cancer (Shen et al. 2012b), cervical cancer cells (Bhushan et al. 2009), prostate cancer cells (Pang et al. 2009), mouse melanoma and human fibrosarcoma cells (Zhao et al. 2003), meningioma cells (Park et al. 2002) DMBA-initiated mice (Huang et al. 2000), mouse colorectal cancer tumors (Yadav et al. 2012), heterotopic (subcutaneous) human pancreatic cancer xenograft nude mouse (Ni et al. 2012), Ehrlich ascitic tumor, Ehrlich ascitic carcinoma and sarcoma-180 tumor model (Qurishi et al. 2013), orthotopic pancreatic cancer nude mouse (Park et al. 2011b), azoxymethane/dextran sodium sulfate-induced colon cancer (Chou et al. 2017) (continued) (Jie et al. 2014), antiangiogenic activity ), decrease in DNA damage and micronucleus formation (Lamy et al. 2012), involvement of Bax/Bcl2, ANLN and S100P pathways (Savio et al. 2015), risk of tumorigenesis (Shukla and Arora 2003) (continued) (Lin et al. 2013) Antimitotic activity via inhibition of tubulin polymerization (Bhattacharyya, et al. 2008), endoplasmic reticulum stress and mitochondrial dysfunction by production of ROS (Kumar et al. 2016a;Larocque et al. 2014 mechanistic studies regarding the safety and efficacy of the newly introduced drugs should be considered to provide better support for the clinical evaluation of these agents. Furthermore, isolated phytochemicals from natural sources can be assessed as new backbones for chemical modifications to design antineoplastic agents with higher efficacy and fewer adverse effects. ...
Cancer has always been considered as a life-threatening disease with an etiology which is not yet completely understood. Currently, there are a wide variety of chemotherapeutic and biological agents in the market which are used for the treatment of different types of cancer under specific guidelines. A numerous number of these chemotherapeutic agents like taxanes, vinca alkaloids, and camptothecin derivatives have been isolated from natural sources; thus, nature provides an endless source of new phytochemicals as lead compounds for the generation of future anticancer agents. To find better candidates as anticancer agents, folklore and traditional medicine of countries all over the world can be a reliable guide. In addition to purification of anticancer compounds, complementary and alternative medicine (CAM) can be considered as an adjuvant therapy in line with the standard chemotherapy protocols (Farzaei et al. in Curr Pharm Des 22:4201–4218, 2016a). To achieve this goal, enough evidence must be provided regarding the safety and efficacy of these natural agents. Traditional Persian medicine (TPM) is one of the most ancient categories of CAM which is globally well-known due to the unique medical texts like Canon of medicine by Avicenna written in 1025 A.D and Razi’s “continens” (Alhavi) written around 960 A.D and its specific doctrine based on temperaments and humors. This chapter aims to introduce the most important anticancer agents used in TPM and summarizing current evidence on the anticancer properties of these natural medicines.
... Bax and Bcl-2 have received attention owing to their regulation of apoptosis which may be understood via their protein expression and therefore the induction of Bax and pro-apoptotic protein expression or inhibition of Bcl-2 and anti-apoptotic protein expression is an important approach for activation of apoptosis in cancer cells (Yip and Reed 2008). Several studies reported that AITC administration upregulated Bax and pro-apoptotic protein expression and downregulated Bcl-2 and anti-apoptotic protein expression in human breast cancer MCF-7 and MDA-MB-231 cells (Bo et al. 2016), human breast adenocarcinoma MDA-MB-468 cells (Tsai et al. 2012), mouse mammary carcinoma EAT cells (Kumar et al. 2009), human brain malignant glioma GBM 8401 cells , human prostate cancer PC-3 and LNCaP cells , human cervical cancer HeLa cells (Qin et al. 2018), human bladder cancer RT4 and T24 cells (Sávio et al. 2015), and renal carcinoma GRC-1 cells (Jiang et al. 2016). Moreover, AITC promoted mitochondrial-mediated apoptotic pathways through AITC-enhanced activities of caspase-3 and caspase-9 in Akt/mTOR signaling pathway in human oral cancer CAR cells (Chang et al. 2021). ...
Cancer growth is a molecular mechanism initiated by genetic and epigenetic modifications that are involved in cell proliferation, differentiation, apoptosis, and senescence pathways. Chemoprevention is an important strategy for cancer treatment that leads to blocking, reversing, or impeding the multistep process of tumorigenesis, including the blockage of its vital morphogenetic milestones viz. normal, preneoplasia, neoplasia, and metastasis. Naturally occurring phytochemicals are becoming ever more popular compared to synthetic drugs for many reasons, including safety, bioavailability, efficacy, and easy availability. Allyl isothiocyanate (AITC) is a natural compound present in all plants of the Cruciferae family, such as Brussels sprouts, cauliflower, mustard, cabbage, kale, horseradish, and wasabi. In vitro and in vivo studies carried out over the decades have revealed that AITC inhibits tumorigenesis without any toxicity and undesirable side effects. The bioavailability of AITC is exceedingly high, as it was reported that nearly 90% of orally administered AITC is absorbed. AITC exhibits multiple pharmacological properties among which its anticancer activity is the most significant for cancer treatment. Its anticancer activity is exerted via selective modulation of multiple cell signaling pathways related to oxidative stress, inflammation, cell proliferation, cell cycle arrest, apoptosis, angiogenesis, invasion, and metastasis. This review highlights the current knowledge on molecular targets that are involved in the anticancer effect of AITC associated with (i) inhibition of carcinogenic activation and induction of antioxidants, (ii) suppression of pro-inflammatory and cell proliferative signals, (iii) induction of cell cycle arrest and apoptosis, and (iv) inhibition of angiogenic and invasive signals related to metastasis.
... Allyl glucosinolate (aka sinigrin) is biologically inactive but hydrolyzed enzymatically in the presence of myrosinase to generate allyl isothiocyanate, which is bioactive and has pharmacological properties [9]. Recent studies have shown that allyl isothiocyanate could induce cell apoptosis and arrest cell cycle in several malignancies [10][11][12][13][14][15][16][17]. ...
Sinigrin is one type of glucosinolates present in cruciferous plants which can be enzymatically hydrolyzed by myrosinase (MYR) to produce allyl isothiocyanate (AITC) with anti-neoplastic potency. This study demonstrates the anticancer activity of silver nanoparticles (AgNPs) green synthesized using sinigrin as a reducing and capping agent. The synthesized sinigrin-coated silver nanoparticles (SIN-AgNPs) characterized by analytical instruments (UV–Vis spectrometry, dynamic light scattering, transmission electron microscopy, and Raman spectroscopy) reveal a mean size of ⁓20 nm with good polydispersity, zeta-potential of -42 mV, and functional groups of sinigrin anchored on the surface of AgNPs. The cDNA of myrosinase was cloned in pcDNA3-eGFP plasmid and transfected to A549 lung adenocarcinoma cells to overexpress myrosinase. The average IC50 values of SIN-AgNPs against parental and myrosinase-expressed A549 cells using the MTT assay were determined to be 3 and 20 µg/ml, respectively. The apoptotic level of myrosinase-expressed A549 cells treated with SIN-AgNPs was further confirmed to be higher than the parental A549 cells.
... After treatment with chalcone 7, in addition to the reduction in cell density, it was possible to observe the presence of elongated cells after treatment with chalcone 7 (Fig. 4). This morphological change has been found associated with reduced cell proliferation, 26,27 which corroborates our results of cytotoxicity and clonogenic assays. In the same way, doxorubicin was used as a positive control and reduction in cell density characteristics of cell death was observed after treatment. ...
... Glucosinolate-derived isothiocyanates (ITCs) showed inhibitory effects on colorectal cancer cell line K562, particularly with ITCs from epi-progoitrin, glucocheirolin, sinigrin, progoitrin, and glucotropaeolin (Leoni et al. 1997). The isothiocyanate family encompasses at least four major members from Cruciferous vegetables, i.e., allyl-isothiocyanate, which inhibits cell growth in RT4 urinary bladder cancer cells and T24 urinary bladder transitional cell line (Savio, da Silva, and Salvadori 2015) as well as impairs the reparation of DNA in human breast cancer cells (Liao et al. 2021), sulforaphane, benzyl-isothiocyanate, and phenethyl-isothiocyanate (Mastuo et al. 2020). Fundamentally, isothiocyanates are biochemical desulfurated byproducts of gluconosilates, principally coming via the activity of a myrosinase ( Figure 1) (Ishida et al. 2014). ...
The use of natural bioactive constituents from various food sources for anticancer purposes has become increasingly popular worldwide. Broccoli (Brassica oleracea var. italica) is on the top of the consumed vegetables by the masses. Its raw matrix contains a plethora of phytochemicals, such as glucosinolates and phenolic compounds, along with rich amounts of vitamins, like vitamin C and minerals. Consumption of broccoli-derived phytochemicals provides strong antioxidant effects, particularly due to its sulforaphane content, while modulating numerous molecules involved in cell cycle regulation, control of apoptosis, and tuning enzyme activity. Thus, the inclusion of broccoli in the daily diet lowers the susceptibility to developing cancers. Numerous studies have underlined the undisputable role of broccoli in the diet as a chemopreventive raw food, owing to the content in sulforaphane, an isothiocyanate produced as a result of hydrolysis of precursor glucosinolates called glucoraphanin. This review will provide evidence supporting the specific role of broccoli and its key bioactive constituents in the prevention and treatment of cancer; a number of studies carried out in the in vitro and in vivo conditions as well as clinical trials were analyzed.
... Consequently, their use in diets and gastronomy has been popularized due to their visual attractiveness and intense flavour as a decorative complement, but also in response to their nutritional quality as a rich source of glucosinolates. Among them, allyl isothiocyanate, which is derived from the GSL sinigrin, has proved to show a positive effect against certain cancers [14] and also has biocide potential [15]. ...
This work studies the enhancement of glucosinolates (GSLs) in mustard sprouts as health promoters. Sprouts of Sinapis alba, Brassica nigra, and B. carinata were grown under broad-spectrum, monochromatic blue or red light-emitting diode (LED) lamps, irrigated with 0–100 mM sodium chloride (NaCl), and sprayed with 0–250 µM methyl jasmonate (MeJA) as elicitor. The use of LEDs did not result in increased sprout biomass in any case. The effect of the applied treatments on the GSLs depended on the species and were restricted to Brassica spp. The red LEDs produced an overall increase in GSLs over 500% in B. carinata (from 12 to 81 mg 100 g−1 F.W.), compared to the white broad-spectrum lights, although the highest increase in content was obtained in treated sprouts with 250 µM MeJA (104 an 105 mg 101 g−1 F.W., under the red and blue LEDs, respectively). The combination of blue LEDs, 100 mM NaCl, and 250 µM MeJA enhanced the levels of GLSs in B. nigra to the maximum (81 mg 100 g−1 F.W.). Overall, these results indicate that by modifying the growing conditions for a given sprout, enhancement in the accumulation of GSLs as health promoters is possible. The use of these treatments is a sustainable alternative to genetic modification when looking for bioactive-enriched foods, delivering natural plant foods rich in bioactive ingredients (e.g., glucosinolates). Nevertheless, the response to the treatments varies among species, indicating that treatments will require adjustment across sprouts. Further research continues with producing cruciferous sprouts to obtain GSL-enriched formulas for further studying the effects of their bioavailability and bioactivity on health-promotion.
... It is derived from cruciferous vegetables including Brassica juncea (L.) (Indian mustard), Wasabia jamonica (L.) matsum (wasabi), and Armoracia rusticana (horseradish) [180][181][182]. It has been shown to exert anticancer [183][184][185][186][187][188], anti-microbial [189][190][191], and anti-fungal activities [192][193][194][195], while also protecting against neurological disorders [196,197]. ...
Isothiocyanates are biologically active secondary metabolites liberated via enzymatic hydrolysis of their sulfur enriched precursors, glucosinolates, upon tissue plant disruption. The importance of this class of compounds lies in their capacity to induce anti-cancer, anti-microbial, anti-inflammatory, neuroprotective, and other bioactive properties. As such, their isolation from natural sources is of utmost importance. In this review article, an extensive examination of the various parameters (hydrolysis, extraction, and quantification) affecting the isolation of isothiocyanates from naturally-derived sources is presented. Overall, the effective isolation/extraction and quantification of isothiocyanate is strongly associated with their chemical and physicochemical properties, such as polarity-solubility as well as thermal and acidic stability. Furthermore, the successful activation of myrosinase appears to be a major factor affecting the conversion of glucosinolates into active isothiocyanates.
... Allyl isothiocyanate is a natural bioactive compound found largely in the plants from Cruciferae family and are highlighted to show several pharmacological effects such as neuroprotective (Subedi et al., 2017), anticancer (Bhattacharya et al., 2010), antidiabetic (Sahin et al., 2019), antifungal (Olivier et al., 1999), and antiulcerative colitis (Kim et al., 2018) activities. Many previous literatures highlighted that the allyl isothiocyanate effectively inhibited the proliferation and triggered apoptosis in numerous tumor cells, such as breast, bladder (Savio et al., 2015), lung (Tripathi et al., 2015), colon (Lai et al., 2014), liver (Hwang and Kim, 2009), prostate (Xu et al., 2006), cervical (Qin et al., 2018), and oral (Chang et al., 2021) cancer cells. ...
Background
Nano-based drug delivery systems have shown several advantages in cancer treatment like specific targeting of cancer cells, good pharmacokinetics, and lesser adverse effects. Liver cancer is a fifth most common cancer and third leading cause of cancer-related mortalities worldwide
Objective
The present study focusses to formulate the selenium (S)/chitosan (C)/polyethylene glycol (Pg)/allyl isothiocyanate (AI) nanocomposites (SCPg-AI-NCs) and assess its therapeutic properties against the diethylnitrosamine (DEN)-induced liver cancer in rats via inhibition of oxidative stress and tumor markers
Methodology
The SCPg-AI-NCs were synthesized by ionic gelation technique and characterized by various characterization techniques. The liver cancer was induced to the rats by injecting a DEN (200mg/kg) on the 8th day of experiment. Then DEN-induced rats treated with 10mg/kg of formulated SCPg-AI-NCs an hour before DEN administration for 16 weeks. The 8-hydroxy-2' -deoxyguanosine (8-OHdG) content, albumin, globulin, and total protein were examined by standard methods. The level of glutathione (GSH), vitamin-C & -E, and superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), and glutathione reductase (GR) activities were examined using assay kits. The liver marker enzymes i.e., alanine transaminase (ALT), aspartate tansaminase (AST), γ-glutamyl transaminase (GGT), lactate dehydrogenase (LDH), and alkaline phosphatase (ALP) activities, alpha fetoprotein (AFP) and carcinoembryonic antigen (CEA), Bax, and Bcl-2 levels, and caspase-3&9 activities was examined using assay kits and the liver histopathology was assessed microscopically by hematoxylin and eosin staining method. The effect of formulated SCPg-AI-NCs on the viability and apoptotic cell death on the HepG2 cells were examined using MTT and dual staining assays, respectively
Results
The results of different characterization studies demonstrated the formation of SCPg-AI-NCs with tetragonal shape, narrowed distribution, and size ranging from 390-450nm. The formulated SCPg-AI-NCs treated liver cancer rats indicated the reduced levels of 8-OHdG, albumin, globulin, and total protein. The SCPg-AI-NCs treatment appreciably improved the GSH, vitamin-C & -E contents, and SOD, CAT, GPx, and GR activities in the serum of liver cancer rats. The SCPg-AI-NCs treatment remarkably reduced the liver marker enzyme activities in the DEN-induced rats. The SCPg-AI-NCs treatment decreased the AFP and CEA contents and enhanced the Bax and caspase 3&9 activities in the DEN-induced rats. The SCPg-AI-NCs effectively decreased the cell viability and induced apoptosis in the HepG2 cells
Conclusion
The present findings suggested that the formulated SCPg-AI-NCs remarkably inhibited the DEN-induced liver carcinogenesis in rats. These findings provide an evidence that SCPg-AI-NCs can be a promising anticancer nano-drug in the future to treat the liver carcinogenesis.
... Moreover, AITC, which is another secondary metabolite found abundantly in cruciferous vegetables and mustard seeds, has potential antitumor activity against bladder cancer. The AITC exert antineoplastic activity via classical BAX/ BCL2 pathway and by modulating the activities of cytokines and cytokinesis as revealed by in vitro studies (Sávio et al., 2015). AITC inhibits proliferation of cancer cells and induced morphological alterations such as elongation of membrane and scattered cellular debris. ...
Carotenoids are lipophilic natural yellow to red pigments found in fruits and vegetables. The chemical molecules of these bioactive compounds comprise an extensive chain of conjugated double bonds responsible for the color and their reactivity and antioxidant activity, which are related to several health benefits. The mechanisms of action of the dietary carotenoids will be presented in this chapter considering the existent scientific evidence based on experimental protocols using in vitro assays, animal models, and clinical trials.
... [14][15][16] In addition to the potential therapeutic advantages of external applications of GI and MU, both substances were reported to have cancer preventive and antiproliferative properties when used as dietary agents. [17][18][19] However, little is understood about warming patients with oncological disease with footbaths containing thermoregulatory substances. Moreover, the symptoms of thermal discomfort and their improvement are often underappreciated and neglected by healthcare providers. ...
Objective
To analyze the thermogenic effects of footbaths with medicinal powders in oncological patients (ON) and healthy controls (HC).
Intervention and Outcomes
Thirty-six participants (23 ON, 13 HC; 24 females; 49.9 ± 13.3 years) received 3 footbaths in a random order with cross-over design: warm water only (WA), warm water plus mustard (MU, Sinapis nigra), and warm water plus ginger (GI, Zingiber officinale). Warmth perception of the feet (Herdecke Warmth Perception Questionnaire, HeWEF) at the follow-up (10 minutes after completion of footbaths, t2) was assessed as the primary outcome measure. Secondary outcome measures included overall warmth as well as self-reported warmth (HeWEF) and measured skin temperature (high resolution thermography) of the face, hands and feet at baseline (t0), post immersion (t1), and follow-up (t2).
Results
With respect to the warmth perception of the feet, GI and MU differed significantly from WA ( P’s < .05) with the highest effect sizes at t1 (WA vs GI, d = 0.92, WA vs MU, d = 0.73). At t2, perceived warmth tended to be higher with GI compared to WA ( d = 0.46). No differences were detected between ON and HC for self-reported warmth. With respect to skin temperatures, face and feet skin temperatures of ON were colder (at t0 and t1, 0.42 ≥ d ≥ 0.68) and tended to have diametrical response patterns than HC (ON vs HC: colder vs warmer after MU).
Conclusion
Among adult oncological patients and healthy controls, footbaths with mustard and ginger increased warmth perception of the feet longer than with warm water only. The potential impact of regularly administered thermogenic footbaths over extended periods merits further investigation for the recovery of cancer-related sense of cold.
... To assess cell density after treatment, the cells were seeded into 24-well culture plates (7.0 × 10 4 cells/ well) and treated for 24 h. Subsequently, the cells were removed using trypsin and stained with 0.4% trypan blue [21]. Untreated cells were used as the negative control. ...
The treatment of bladder cancer remains a challenge in clinical practice. Different chemotherapeutic protocols can be used; however, it is common to observe tumor recurrence and secondary effects that result in toxicity. Doxorubicin (DOX), one of the most effective anticancer agents used to treat bladder cancer, can cause chronic cardiotoxicity, limiting its use in clinical practice. Resveratrol (RES), a natural product with potential antitumor activity against bladder cancer, is associated with rapid metabolism and low bioavailability and needs to be combined with chemotherapeutic drugs to improve its use. Our study aimed to assess the therapeutic effect of a low concentration of DOX (2 µM) in combination with RES (150, 200 and 250 µM) on two bladder cancer cell lines. We investigated the mechanism of interaction between the drugs by performing cytotoxicity, clonogenic, oxidative stress, cell migration, cell morphology and nuclear division index (NDI) assays. Cytotoxicity evaluation revealed an additive interaction between RES and DOX for both cell lines. Additionally, the results of cell colony formation, oxidative stress, cell migration, cell morphology and NDI assays showed that a combination of DOX and RES was more effective than RES or DOX alone. In conclusion, a low concentration of DOX combined with RES could potentiate the antitumor effects of the drugs on bladder cancer cells, thus overcoming the secondary effects caused by DOX and the low bioavailability of resveratrol.
... On the other hand, two cruciferous extracts (from mustard seeds and wall rocket) that are rich in sinigrin [36,57,58,61] were found to exert a strong inhibition of SARS-CoV-2 3CL Pro activity. Sinigrin is the precursor glucosinolate of allyl isothiocyanate, a reactive molecule displaying antimicrobial [82,83] and anticarcinogenic properties [84][85][86]. By contrast, references to the potential of either sinigrin or allyl isothiocyanate as antivirals are more limited [16]. ...
Antiviral treatments inhibiting Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) replication may represent a strategy complementary to vaccination to fight the ongoing Coronavirus disease 19 (COVID-19) pandemic. Molecules or extracts inhibiting the SARS-CoV-2 chymotripsin-like protease (3CLPro) could contribute to reducing or suppressing SARS-CoV-2 replication. Using a targeted approach, we identified 17 plant products that are included in current and traditional cuisines as promising inhibitors of SARS-CoV-2 3CLPro activity. Methanolic extracts were evaluated in vitro for inhibition of SARS-CoV-2 3CLPro activity using a quenched fluorescence resonance energy transfer (FRET) assay. Extracts from turmeric (Curcuma longa) rhizomes, mustard (Brassica nigra) seeds, and wall rocket (Diplotaxis erucoides subsp. erucoides) at 500 µg mL−1 displayed significant inhibition of the 3CLPro activity, resulting in residual protease activities of 0.0%, 9.4%, and 14.9%, respectively. Using different extract concentrations, an IC50 value of 15.74 µg mL−1 was calculated for turmeric extract. Commercial curcumin inhibited the 3CLPro activity, but did not fully account for the inhibitory effect of turmeric rhizomes extracts, suggesting that other components of the turmeric extract must also play a main role in inhibiting the 3CLPro activity. Sinigrin, a major glucosinolate present in mustard seeds and wall rocket, did not have relevant 3CLPro inhibitory activity; however, its hydrolysis product allyl isothiocyanate had an IC50 value of 41.43 µg mL−1. The current study identifies plant extracts and molecules that can be of interest in the search for treatments against COVID-19, acting as a basis for future chemical, in vivo, and clinical trials.
... Allyl isothiocyanate is a strong, volatile, color-less oil that is found in Brassica plants of the Cruciferae family (Sharma et al., 2012). Allyl isothiocyanate is found to suppress the viability, multiplication, invasion, and to trigger the apoptotic cell deaths in the different human cancer cells like liver, cervical, and bladder (Zhang, 2010;Hwang and Lee, 2006;Qin et al., 2018;Savio et al., 2015). Furthermore, the effectiveness of Allyl isothiocyanate in combination with radiation therapy is most effective against the non-small cell lung cancer cells (Tripathi et al., 2015). ...
Background
Colorectal cancer (CRC) is a frequently diagnosed cancer that primarily affects the digestive system and an imperative cause of mortalities worldwide.
Objective
In this work, we formulated the Tin oxide-Sodium alginate-Polyethylene glycol-Allyl isothiocyanate nanocomposites (SAP-Ally-NCs) and investigated its anticancer role against the DMH-provoked CRC in rats.
Methodology
The formulated SAP-Ally-NCs were characterized different techniques. The CRC was provoked to the rats via injecting 20mg/kg of DMH and then administered with the formulated SAP-Ally-NCs for 16 weeks. The bodyweight changes and the polyp’s incidences were detected and tabulated. The status of lipid peroxidation and antioxidant enzymes were studied by standard techniques. The inflammatory markers and xenobiotic enzymes level was scrutinized using respective kits. The mRNA expressions of various signaling molecules were examined by RT-PCR. The liver and colon tissues were examined microscopically to detect the histological changes.
Results
The formulated SAP-Ally-NCs treatment appreciably improved the body weight gain and suppressed the polyp’s incidences in the DMH-challenged animals. SAP-Ally-NCs treated animals were demonstrated the notable reduction in the lipid peroxidation and inflammatory cytokines and elevated the antioxidant enzymes i.e. CAT and SOD activity. SAP-Ally-NCs administered animals exhibited the noticeable reduction in the expression of PCNA, cyclin-D1, iNOS, and COX-2 in the colon tissues. The histological findings also unveiled the therapeutic role of SAP-Ally-NCs.
Conclusion
In conclusion, the SAP-Ally-NCs demonstrated the potent anticancer action against the DMH-provoked CRC in rats. In future, it could be a potent chemotherapeutic agent to the CRC.
... 83 In addition to phenolics, the presence of the glucosinolate sinigrin was reported to take part in the inhibition of hepatocarcinogenesis. 85 Moreover, mustard extracts rich in allyl isothiocyanate were reported to possess cytotoxic effects against bladder cancer cells. 86,87 The presence of allyl isothiocyanate, together with phenyl isothiocyanate, in mustard extracts has been found to exhibit anti-inflammatory effects by inhibiting some inflammationpromoting cytokines such as interleukin-6 (IL-6) and tumor necrosis factor α (TNF-α). 88 Overall, laser light treatment induced the antioxidant activity and the anticancer activity as indicated by increased QR and GST enzymes activities in all the studied Brassica sprouts. ...
... After 24 h, cells were treated with the chrysin at concentrations of 20, 40 and 60 μM for 24 h. Subsequently, cells were removed using trypsin and stained with 0.4% Trypan Blue (25). Untreated cells were used as a negative control. ...
The antitumour activity of chrysin have been studied in several types of cancer cells. In urinary bladder cancer, its cytotoxic effects have already demonstrated; however, its mechanism of action is not completely understood and the role of tumour protein p53 (TP53) gene in these effects is unclear. In this study, we investigated the role of chrysin (10, 20, 40, 60 80 and 100 µM) in progression of bladder tumour cells with different status of the TP53 gene and different degrees of tumour (RT4, grade 1, TP53 wild type; 5637, grade 2, TP53 mutated and T24, grade 3, TP53 mutated). Results demonstrated that chrysin inhibited cell proliferation by increasing reactive oxygen species and DNA damage and inhibited cell migration in all cell lines. In TP53 wild-type cells, a sub-G1 apoptotic population was present. In mutated TP53 cells, chrysin caused arrest at the G2/M phase and morphological changes accompanied by downregulation of PLK1, SRC and HOXB3 genes. In addition, in Grade 2 cells, chrysin induced global DNA hypermethylation and, in the highest-grade cells, downregulated c-MYC, FGFR3 and mTOR gene expression. In conclusion, chrysin has antiproliferative and toxicogenetic activity in bladder tumour cells independently of TP53 status; however, the mechanisms of action are dependent on TP53 status.
... 33 AITC is one of common ITCs which was reported to have potential chemopreventive and anticancer properties on human cancer cells of hepatoma, bladder cancer, and non-small cell lung cancer. 15,18,20,21 Nevertheless, little is known about its effect on oral cancer cells, especially in a cisplatin-resistant cancer status. In our study, AITC inhibited cell proliferation and induced apoptosis in CAR cells. ...
Background/purpose
Cisplatin-resistant oral cancer is clinically difficult to manage and the dose-dependent toxicities of cisplatin has been widely concerned. Allyl isothiocyanate (AITC), known as mustard oil, is a plant-derived compound abundant in cruciferous vegetables. It is reported to have anti-cancer potential as a natural dietary chemopreventive compound against a variety of cancers, but the effect of AITC on cisplatin-resistant cancer cells is still little-known.
Methods
Human CAL27-cisplatin-resistant oral cancer cells (CAR cells) were examined to investigate the antitumor properties of AITC. 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide (MTT) assay, IncuCyte™ S3 cell proliferation assay, 4′,6-diamidino-2-phenylindole (DAPI) and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining as well as Western blot analysis were deployed.
Results
AITC decreased CAR cell viability, induced cell death of CAR cells and inhibited the confluences of cultured CAR cells. When CAR cells were treated with AITC, activation of caspase-3 and caspase-9 by AITC was observed and could be reversed by Z-VAD-fmk (pan-caspase inhibitor). Furthermore, the protein expressions of phosphorylated protein kinase B (p-AKT) and phosphorylated mammalian target of rapamycin (p-mTOR) were suppressed in AITC-treated CAR cells, whereas protein expressions of Bax, cytochrome c, Apaf-1, cleaved caspase-3, and cleaved caspase-9 were upregulated in AITC-treated CAR cells.
Conclusion
AITC can inhibit Akt/mTOR proliferation signaling and promote mitochondria-dependent apoptotic pathway through AITC-enhanced activities of caspase-3 and caspase-9 in CAR cells.
... Consequently, its accumulation in high levels can be considered greatly responsible for the characteristic flavor of wall rocket. Furthermore, allyl ITC has been also investigated for its potential health benefits, mainly as antitumoral agent [14][15][16]. Therefore, including vegetables that are source of allyl ITC in our diet may have a positive effect in increasing health status. ...
Wall rocket is a leafy vegetable with pungent flavor related to the presence of isothiocyanates (ITCs). Despite interest in it as a crop of high organoleptic quality, the variability of the volatile profile in the species remains unknown. Twenty-four populations grown under a greenhouse were evaluated. A considerable diversity for the total levels of volatiles was found, providing information of the aroma intensity among accessions. ITCs represented the main fraction. Allyl ITC was the main compound, and levels showed up to 6-fold difference among populations. The esters fraction was mainly represented by cis-3-hexenyl isovalerate and cis-3-hexenyl butyrate, with 20-fold differences among populations. Additionally, the content in sinigrin was evaluated as main GSL in wall rocket. Differences reached up to 13-fold. These results suggest that some populations can be used to develop highly pungent varieties, whereas some others can be selected for mild-pungent varieties, as it is the case of DER045 with low levels of ITCs and high in esters. The presence of several ITCs in the profile also suggested the presence of other novel GSLs. Overall, the work increases the knowledge in the variability of wall rocket for the volatile profile and sinigrin accumulation, a starting point for future breeding programs.
... An extract of B. nigra has been able to protect HepG2 cells against benzo[a]pyrene-induced DNA damage, possibly via mechanisms associated with induction of detoxification enzymes [23]. The cytotoxic effects of allyl isothiocyanate, present in mustard seeds, have been reported against lung [24] and bladder cancer cells [25,26]. Moreover, previous studies reported that dietary mustard seeds suppressed azoxymethane-induced colon adenomas in mice [27] and dimethylhydrazine-induced colorectal carcinomas in rats [28]. ...
Lung cancer is the primary cause of cancer-related death worldwide, and development of novel lung cancer preventive and therapeutic agents are urgently needed. Brassica nigra (black mustard) seeds are commonly consumed in several Asian and African countries. Mustard seeds previously exhibited significant anticancer activities against several cancer types. In the present study, we have investigated various cellular and molecular mechanisms of anticancer effects of an ethanolic extract of B. nigra seeds against A549 and H1299 human non-small cell lung cancer cell lines. B. nigra extract showed a substantial growth-inhibitory effect as it reduced the viability and clonogenic survival of A549 and H1299 cells in a concentration-dependent manner. B. nigra extract induced cellular apoptosis in a time- and concentration-dependent fashion as evidenced from increased caspase-3 activity. Furthermore, treatment of both A549 and H1299 cells with B. nigra extract alone or in combination with camptothecin induced DNA double-strand breaks as evidenced by upregulation of γH2A histone family member X, Fanconi anemia group D2 protein, Fanconi anemia group J protein, ataxia‑telangiectesia mutated and Rad3-related protein. Based on cell cycle analysis, B. nigra extract significantly arrested A549 and H1299 cells at S and G2/M phases. Additionally, B. nigra extract suppressed the migratory and invasive properties of both cell lines, downregulated the expression of matrix metalloproteinase-2 (MMP2), MMP9, and Snail and upregulated the expression of E-cadherin at mRNA and protein levels. Taken together, these findings indicate that B. nigra seed extract may have an important anticancer potential against human lung cancer which could be mediated through simultaneous and differential regulation of proliferation, apoptosis, DNA damage, cell cycle, migration, and invasion.
... 115,116 3-butenyl isothiocyanate isolated from B. juncea was found to be effective against the human cancer prostrate cell lines by inducing apoptosis. 117 AITC isolated from mustard seeds inhibited cell proliferation and reduced bladder cancer in two cell lines by two different mechanisms; in RT4 cells, it mainly acted via the classical Bcl-2 associated X/Bcl-2 pathway, while in T24 cells, AITC modulated the activities of anillin actin binding protein (related to cytokinesis) and S100 calcium binding protein P. 118 Effect on platelet function/cardiovascular activity Cumin and fennel inhibited platelet aggregation induced by arachidonic acid, collagen, etc. Cumin inhibited thromboxane B2 production and increased the formation of lipoxygenase-derived products. 119 Decreased paraxonase activity is linked with heart diseases and cumin increased the activity of paraxonase in patients with high cholesterol. ...
... Savio and colleagues also showed that while AITC affected the BAX/BCL2 pathway in RT4 cells (wt TP53), AITC affected the cytokinesis related ANLN and S100P in T24 (mutant TP53) cells. This suggests AITC regulates bladder cancer cell gene expression in a TP53 genotype-dependent manner [141]. ...
Bladder cancer is a significant health burden due to its high prevalence, risk of mortality, morbidity, and high cost of medical care. Epidemiologic evidence suggests that diets rich in cruciferous vegetables, particularly broccoli, are associated with lower bladder cancer risk. Phytochemicals in cruciferous vegetables, such as glucosinolates, which are enzymatically hydrolyzed to bioactive isothiocyanates, are possible mediators of an anticancer effect. In vitro studies have shown inhibition of bladder cancer cell lines, cell cycle arrest and induction of apoptosis by these isothiocyanates, in particular sulforaphane and erucin. Although, not yet completely understood, many mechanisms of anti‐cancer activity at the steps of cancer initiation, promotion and progression have been attributed to these isothiocyanates. They target multiple pathways including the adaptive stress response, phase I/II enzyme modulation, pro‐growth, ‐survival, ‐inflammatory signaling, angiogenesis, and even epigenetic modulation. Multiple in vivo studies have shown the bioavailability of isothiocyanates and their anti‐tumoral effects. Although human studies are limited, they support oral bioavailability with reasonable plasma and urine concentrations achieved. Overall, both cell and animal studies support a potential role for isothiocyanates in bladder cancer prevention and treatment. Future studies are necessary to examine clinically relevant outcomes and define guidelines on ameliorating the bladder cancer burden.
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... CDK1 regulates the G1-S transition in the cell cycle, a process that is important for the development of centrosome mutation (48). CDK1 is a vital regulator in cell proliferation, and overexpression may lead to high tumor aggressiveness and poor prognosis (49)(50)(51). Some antibodies, including anti-CDK1, have been used to investigate cell proliferation (52). ...
The purpose of the present study was to identify key genes and investigate the related molecular mechanisms of bladder cancer (BC) progression. From the Gene Expression Omnibus database, the gene expression dataset GSE7476 was downloaded, which contained 43 BC samples and 12 normal bladder tissues. GSE7476 was analyzed to screen the differentially expressed genes (DEGs). Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were performed for the DEGs using the DAVID database, and a protein‑protein interaction (PPI) network was then constructed using Cytoscape software. The results of the GO analysis showed that the upregulated DEGs were significantly enriched in cell division, nucleoplasm and protein binding, while the downregulated DEGs were significantly enriched in 'extracellular matrix organization', 'proteinaceous extracellular matrix' and 'heparin binding'. The results of the KEGG pathway analysis showed that the upregulated DEGs were significantly enriched in the 'cell cycle', whereas the downregulated DEGs were significantly enriched in 'complement and coagulation cascades'. JUN, cyclin‑dependent kinase 1, FOS, PCNA, TOP2A, CCND1 and CDH1 were found to be hub genes in the PPI network. Sub‑networks revealed that these gene were enriched in significant pathways, including the 'cell cycle' signaling pathway and 'PI3K‑Akt signaling pathway'. In summary, the present study identified DEGs and key target genes in the progression of BC, providing potential molecular targets and diagnostic biomarkers for the treatment of BC.
... AITC was found to inhibit cell proliferation independent of TP53 status. AITC acted via the classical BAX/BCl2 pathway in RT4 cells, while in T24 cells, AITC modulated the activities of ANLN that is related to cytokinesis and S100P [31][32][33][34]. ...
Background:
Brassica nigra belonging to the family Brassicaceae (syn - black mustard) comprises of dried seeds and is widely cultivated in Mediterranean region and various other countries like India and Europe. The Plant is neither reported in any official book nor reported systematically for pharmacological effects.
Methods:
We undertook a structured search of all the official books including pharmacopoeias and ayurvedic textbooks. The data was collected, sorted and compiled to bring the chemical constituents and uses of B. nigra.
Results:
B. nigra contains glycosinolates which undergoes hydrolysis in the presence of myrosinase enzyme yields allyl isothiocyante. The bitter taste and pungent odor is due to the isothiocyanates. B. Nigra shows various activities that are medically important such as anticancer, anti-bacterial, antifungal, anti-helminthic, protection against renal and hepatic toxicity and can also be used in diabetes treatment. Black mustards are also used in cardiovascular and neurological disorders.
Conclusion:
The present review explores the historical background including the macroscopic and microscopic characteristics, chemical constituents, pharmacological action, various evaluation parameters, formulations and uses of black mustard.
... The mode of action for the chemopreventive activity of AITC is attributed primarily to the detoxification of carcinogens through activation of nuclear factor erythroid-related factor2 (Nrf2) 4 . AITC also inhibited the growth of various human cancer cell lines such as colorectal carcinoma 5 , lung cancer 6 , leukemia 7 , breast adenocarcinoma 8 , bladder cancer 3,9 , neuroblastoma 10 , hepatoma 11 and prostate cancer cells 2,12 . The mechanisms are likely to involve DNA damage 6 , cell cycle arrest and apoptosis 8,12,13 and binding to thiol-reactive groups of several cellular targets such as DNA topoisomerase 2, p53 and tubulins 4,14,15 . ...
Allyl isothiocyanate (AITC), a dietary phytochemical in some cruciferous vegetables, exhibits promising anticancer activities in many cancer models. However, previous data showed AITC to have a biphasic effect on cell viability, DNA damage and migration in human hepatoma HepG2 cells. Moreover, in a 3D co-culture of HUVEC with pericytes, it inhibited tube formation at high doses but promoted this at low doses, which confirmed its biphasic effect on angiogenesis. siRNA knockdown of Nrf2 and glutathione inhibition abolished the stimulation effect of AITC on cell migration and DNA damage. The biological activity of a novel AITC-conjugated silicon quantum dots (AITC-SiQDs) has been investigated for the first time. AITC-SiQDs showed similar anti-cancer properties to AITC at high doses while avoiding the low doses stimulation effect. In addition, AITC-SiQDs showed a lower and long-lasting activation of Nrf2 translocation into nucleus which correlated with their levels of cellular uptake, as detected by the intrinsic fluorescence of SiQDs. ROS production could be one of the mechanisms behind the anti-cancer effect of AITC-SiQDs. These data provide novel insights into the biphasic effect of AITC and highlight the application of nanotechnology to optimize the therapeutic potential of dietary isothiocyanates in cancer treatment.
... Moreover, studies have shown that AITC displays anticancer activity. AITC inhibited proliferation through induction of apoptosis and cell cycle arrest in breast [16,17] and bladder [18,19] cancer cells as well as in brain malignant glioma [20], suppressed epidermal growth factor-stimulated invasion and migration in colorectal adenocarcinoma cells [21], inhibited proliferation via induction of apoptosis and cell cycle arrest in prostate cancer cells in vitro [22] and in vivo [23], and induced apoptosis in leukemia cells [24]. In addition to these studies, AITC has been reported to sensitize ovarian and lung cancer cells to chemotherapeutic drug cisplatin [25], and to exert synergistic therapeutic effects on lung cancer cells in combination with radiation [26]. ...
It was reported recently that allyl isothiocyanate (AITC) could inhibit various types of cancer cell growth. In the present study, we further investigated whether AITC could inhibit the growth of human breast cancer cells. Unexpectedly, we found that AITC did not inhibit, rather slightly promoted, the proliferation of MDA-MB-231 breast cancer cells, although it did have inhibitory effect on MCF-7 breast cancer cells. Cytofluorimetric analysis revealed that AITC (10 µM) did not induce apoptosis and cell cycle arrest in MDA-MB-231 cells. In addition, AITC significantly (p < 0.05) increased the expression of BCL-2 and mTOR genes and Beclin-1 protein in MDA-MB-231 cells. No significant changes in expression of PRKAA1 and PER2 genes, Caspase-8, Caspase-9, PARP, p-mTOR, and NF-κB p65 proteins were observed in these AITC-treated cells. Importantly, AITC displayed cytotoxic effect on MCF-10A human breast epithelial cell line. These observations suggest that AITC may not have inhibitory activity in MDA-MB-231 breast cancer cells. This in vitro study warrants more preclinical and clinical studies on the beneficial and harmful effects of AITC in healthy and cancer cells.
... Recent studies showed that natural bioactive compounds, including isothiocyanate and amygdalin, have anticancer effects on bladder cancer cells (5,6). Garlic, a species of the Allium genus, has been utilized for medicinal purposes in recorded history (7)(8)(9). ...
To examine the anticancer effects of S-allyl-L-cysteine (SAC) in human bladder cancer cells and to identify possible molecular mechanisms, bladder cancer cell lines (HTB5, HTB9, JON, UMUC14, T24, and cisplatin resistant-T24R2) were incubated with SAC, and cell proliferation was measured using the Cell Counting Kit-8 assay and clonogenic assay. Cell cycle and apoptosis were evaluated by flow cytometry. Expression levels of apoptosis- and cell cycle-associated proteins were analyzed by western blotting. Proliferation and colony formation in bladder cancer cells was significantly inhibited by SAC treatment in a dose-dependent manner. SAC treatment significantly enhanced apoptosis and promoted a cell cycle arrest in the S phase. SAC also increased the expression of apoptosis-related genes, including caspases, poly (ADP-ribose) polymerase and cytochrome c. SAC had an anticancer effect on bladder cancer cells in vitro, at least partially, through the induction of apoptosis and a cell cycle arrest. SAC is a potential therapeutic agent for the treatment of bladder cancer.
Myocardial infarction (MI) is a common type of ischemic heart disease that affects millions of people worldwide. In recent times, nanotechnology has become a very promising field with immense applications. The current exploration was conducted to synthesize the chitosan‐sodium alginate‐polyethylene glycol‐Ally isothiocyanate nanocomposites (CSP‐AIso‐NCs) and evaluate their beneficial roles against the isoproterenol (ISO)‐induced MI in rats. The CSP‐AIso‐NCs were prepared and characterized by several characterization techniques. The MI was initiated in the rats by the administration of 85 mg/kg of ISO for 2 days and treated with 10 and 20 mg/kg of CSP‐AIso‐NCs for 1 month. The changes in heart weight and bodyweight were measured. The cardiac function markers were assessed with echocardiography. The lipid profiles, Na+, K+, and Ca ²⁺ ions, cardiac biomarkers, antioxidant parameters, and inflammatory cytokines were assessed using corresponding assay kits. The histopathological study was done on the heart tissues. The UV spectral analysis revealed the maximum peak at 208 nm, which confirms the formation of CSP‐AIso‐NCs. The FT‐IR analysis revealed the occurrence of different functional groups, and the crystallinity of the CSP‐AIso‐NCs was proved by the XRD analysis. DLS analysis indicated the size of the CSP‐AIso‐NCs at 146.50 nm. The CSP‐AIso‐NCs treatment increased the bodyweight and decreased the HW/BW ratio in the MI rats. The status of lipids was reduced, and HDL was elevated in the CSP‐AIso‐NCs administered to MI rats. CSP‐AIso‐NCs decreased the LVEDs, LVEDd, and NT‐proBNP and increased the LVEF level. The oxidative stress markers were decreased, and the antioxidants were increased by the CSP‐AIso‐NCs treatment in the MI rats. The Na+ and Ca+ ions were reduced, and the K+ ions were increased by the CSP‐AIso‐NCs. The interleukin‐1β and tumor necrosis factor‐α were also depleted, and Nrf‐2 was improved in the CSP‐AIso‐NCs administered to MI rats. The histological study revealed the ameliorative effects of CSP‐AIso‐NCs. Overall, our outcomes revealed that the CSP‐AIso‐NCs are effective against the ISO‐induced MI rats. Hence, it could be a hopeful therapeutic nanomedicine for MI treatment.
Over the years, extensive research on mustard oil has revealed the presence of isothiocyanates as pungent toxic compounds and erucic acids as possible health risks. Hence, strategies are needed to remove or reduce the isocyanates and erucic contents while processing the mustard oil. Conversely, due to the presence of monounsaturated fatty acids and omega-3 fatty acids, e.g., erucic acid and α-linolenic acid, mustard oil possesses hypolipidemic effect. The oil rich in erucic acid has demonstrated anti-inflammatory and antioxidant effect that may help in the prevention of obesity-induced metabolic disorders, melanoma, and the Parkinson’s disease. The hypolipidemic effects further potentiate the antidiabetic effect. The presence of allyl isothiocyanate has promising anticancer evidenced by the cell cycle arrest at the G2/M phase. Mustard oil chemical compounds showed antimicrobial action against several pathogenic microorganisms and proved to be useful for active antimicrobial packaging. A few of the studies also reported its antioxidant and antiviral activities. Studies confirmed that allyl isothiocyanate from mustard oil has anti-inflammatory potential when applied topically. The economically motivated adulteration of mustard oil with argemone needs further analytical investigation. Further consideration of the mustard oil benefits-risk ratio and mustard oil optimization are required to improve its health-boosting properties.Graphical Abstract
Allyl-isothiocyanate (AITC) is a common Isothiocyanates (ITC) and its chemo-preventive and anti-tumor effects are believed to be related to the activation of NF-E2 p45-related Factor 2 (Nrf2). However, its anti-tumor effects on colorectal cancer (CRC) are not well elucidated. Here, we investigated the therapeutic in vitro and/or in vivo effects and mechanisms of action (MOA) for AITC on CRC cell line HCT116 (human) and MC38 (mouse). AITC treatment in a low concentration range (1 mg/kg in vivo) significantly inhibited the tumor cell growth and increased the expression of p21 and Nrf2. The AITC-mediated induction of p21 was dependent on Nrf2 but independent on p53 in vitro and in vivo at low dose. In contrast, the high dose of AITC (5 mg/kg in vivo) failed to increase substantial levels of p21/MdmX, and impaired the total antioxidant capacity of tumors and subsequent anti-tumor effect in vivo. These results suggest that an optimal dose of AITC is important and required for the proper Nrf2 activation and its anti-CRC effects and thus, providing insights into the potential applications of AITC for the prevention and treatment of CRC.
Allyl isothiocyanates (AITC) have gained recognition in recent years as effective chemotherapeutic and epigenetic modulators. The chemopreventive properties and toxicological perspectives of AITCs from the last few decades were taken into account by a number of investigations. Their active therapeutic relevance was hindered by a number of factors, including instability under typical physiological conditions and low bioavailability due to low aqueous solubility. In this review, we highlighted the chemopreventive attributes of AITC in relation to its molecular mechanisms and metabolic fate for cancer. Moreover, we emphasized on investigational anticancer activities and various strategies for delivery of AITC in different types of cancer. Considering cellular interactions, we shed light on the toxicological properties of AITCs to address further issues regarding their assessment in therapeutic development. This review identifies knowledge gaps with various contemporary approaches involving most recent studies and may pave the way for a better understanding for the development of novel AITC therapeutics.
Graphical abstract
Sulfur (S) is an essential nutrient assimilated through the diet and incorporated into organic structures such as amino acids, coenzymes, and other bioactive compounds. Plants’ ability to regulate stress resistance via secondary metabolism has extended the interest in S-containing compounds, driven by their additional properties as bioactive molecules. Among plant families that produce S-compounds, the Brassicaceae, which includes broccoli, cabbage, and cauliflower, and the Liliaceae family, which includes garlic and onion, stand out. In recent years, the research has been focused on S-containing amino acids (mainly methionine and cysteine) and glucosinolates (GSLs) and their hydrolysis products like isothiocyanates but also in other S-containing compounds such as phytoalexins or cysteine sulfoxides. GSLs are becoming more popular because of their specific biological properties, including antioxidant, anti-inflammatory, or antimicrobial, among others. Accordingly, a diet rich in vegetables containing S-containing compounds has been associated with a lower risk of developing cancer, neurological diseases like Alzheimer’s, inflammatory bowel disease, cardiovascular diseases, several skin disorders, and obesity. Hence, S-metabolites can therefore be used as therapeutic and preventative components in functional foods and nutraceuticals, as well as cosmeceutical products. This chapter aims to revise the most important features related to sulfur metabolism and S-containing compounds from plant sources, with emphasis on their involvement in secondary metabolism, natural sources, structural classification, biological functions, and applications in human nutrition and health.
Allyl isothiocyanate (AITC) is a phytochemical that is abundantly present in cruciferous vegetables of the Brassicaceae family, such as cabbage, broccoli, mustard, wasabi, and cauliflower. The pungent taste of these vegetables is mainly due to the content of AITC present in these vegetables. AITC is stored stably in the plant as its precursor sinigrin (a type of glucosinolate), which is physically separated from myrosin cells containing myrosinase. Upon tissue disruption, myrosinase gets released and hydrolyzes the sinigrin to produce AITC and by-products. AITC is an organosulfur compound, both an irritant and toxic, but it carries pharmacological properties, including anticancer, antibacterial, antifungal, and anti-inflammatory activities. Despite the promising anticancer effectiveness of AITC, its clinical application still possesses challenges due to several factors, i.e., low aqueous solubility, instability, and low bioavailability. In this review, the anticancer activity of AITC against several cancer models is summarized from the literature. Although the mechanism of action is still not fully understood, several pathways have been identified; these are discussed in this review. Not much attention has been given to the delivery of AITC, which hinders its clinical application. However, the few studies that have demonstrated the use of nanotechnology to facilitate the delivery of AITC are addressed.
A wide variety of lactic acid-fermented fruits and vegetables currently exists. Among them, kimchi and sauerkraut are the ones that have met worldwide commercial significance and are most widely consumed. Cabbage is the primary ingredient for both; however, kimchi production involves incorporating many more ingredients, reflected in the complexity of the respective micro-ecosystem. Traditional fermented foods have a long history of being consumed by local communities and are important nutritional and functional properties. Their health functionality includes anticancer, antiobesity, anticonstipation, colorectal health promotion, probiotic properties, cholesterol reduction, fibrinolytic effect, antioxidative, antiaging, brain health promotion, immune promotion, skin health promotion, etc. This chapter presents and critically discusses the various aspects of fermentation and processing along with nutritional and health functionalities of kimchi and sauerkraut.
The present study was performed to evaluate the safe dose of ethanolic Eruca sativa leaves extract in healthy adult male rats. Twenty-five male Wister rats were divided into 5 groups: Group I (Control): received no treatment, groups 2, 3, 4, and 5 received 1, 3, 5, and 6 g/kg BW orally per day for 35 days. On the 36 day, rats were euthanized. Body weights were recorded. Serum biochemical analysis and the expression level of liver and kidney p53 and Bcl-2 were determined. Histopathology of liver and kidney, and TNF-α immunohistochemistry were examined. The ethanolic extract revealed the content of phenols, flavonoids, and fatty acids. Determination of biochemical parameters showed no significant difference in the estimated parameters between control and treated groups. While the treated groups did not show any significant changes expressions of p53 and Bcl-2 as observed by histopathological and immunohistochemical examination as compared to the control group, liver and kidney toxicity occurred for the rat groups administered 6g/kg. In conclusion, Eruca sativa is not toxic and there were no structural and functional changes in the liver and kidney when administered the doses of 1–5 g/Kg BW over a 35-day period through its potent antioxidant activity in rats and that toxicity that occurred in the 6g/kg-day group may be caused by a high concentration of glucosinolates.
This chapter introduces the health benefits of glucosinlonate-isothiocyanates (ITCs), the myrosinase hydrolysis products of sulfur-containing secondary metabolites derived from cruciferous vegetables. The ITCs are known to be safe with little or no side effects and have been widely studied for their bioactivities against communicable and noncommunicable diseases such as neurodegenerative diseases and cancer. Sulforaphane, glucomoringin-isothiocyanate, and allyl-ITC are the widely studied isothiocyanates in recent years, they possess robust medicinal potential against both several noncommunicable diseases. The compounds exert their effects via modulation of crucial signaling pathways and expression of genes or proteins implicated in the process. ITCs influence the upregulation of series protective genes via the activation of nuclear factor erythroid 2 related factor 2, which is a central transcription factor for phase II enzymes upregulation in response to oxidative stress. Therefore the chapter discusses the in vitro and in vivo biological activities of the most studied isothiocyanates and their mechanism of action with particular emphasis on neuroprotection and their ameliorative effect on cancer, oxidative stress, inflammation, and diabetes mellitus.
Mustard seeds are used as a food, and spice due to their nutritive value, the presence of biologically active compounds, and specific taste. Its morphology was investigated by optical microscopy and scanning electron microscopy before and after isolation by hydrodistillation and microwave assisted distillation. The chemical composition of volatile compounds in different mustard seeds (Brassica juncea L. and Brassica nigra L.) was determined using GC–MS, after applying various techniques (Clevenger hydrodistillation, microwave assisted distillation—MAD, and microwave hydrodiffusion and gravity—MHG). Allyl isothiocyanate, degradation product of allyl glucosinolate, was the main volatile compound in B. juncea essential oils and extract after all applied extraction techniques (91.07–99.01%). Gastrointestinal stability of allyl isothiocyanate was determined employing two-phase digestion model (gastric and duodenal) by two methods (in vitro digestion method using commercial and ex vivo digestion method using human digestive enzymes). The stability rate of allyl isothiocyanate was higher after the gastric digestion phase by both methods. In B. nigra seeds but-3-enyl and allyl isothiocyanate were the main compounds after Clevenger hydrodistillation (80.58% and 15.39%, respectively). After MAD, and MHG the main compounds were 4,5-epithiopentanenitrile (50.70% and 59.93%, respectively), and 3,4-epithiobutanenitrile (7.61% and 25.97%, respectively), originating from the same glucosinolates, i.e. but-3-enyl and allyl glucosinolate, respectively. Antiproliferative activity of mustard seed EOs and extracts was evaluated against human cancer cell lines (MDA-MB-231 and TCCSUP). The best antiproliferative activity was shown for B. nigra MHG extract against MDA-MB-231 cell line with IC50 value of 9.1 µg/mL.
Cell cycle, growth, survival and metabolism are tightly regulated together and failure in cellular regulation leads to carcinogenesis. Several signaling pathways like the PI3K, WNT, MAPK and NFKb pathway exhibit aberrations in cancer and help achieve hallmark capabilities. Clinical research and in vitro studies have highlighted the role of epigenetic alterations in cancer onset and development. Altered gene expression patterns enabled by changes in DNA methylation, histone modifications and RNA processing have proven roles in cancer hallmark acquisition. The reversible nature of epigenetic processes offers robust therapeutic targets. Dietary bioactive compounds offer a vast compendium of effective therapeutic moieties. Isothiocyanates (ITCs) sourced from cruciferous vegetables demonstrate anti-proliferative, pro-apoptotic, anti-inflammatory, anti-migratory and anti-angiogenic effect against several cancers. ITCs also modulate the redox environment, modulate signaling pathways including PI3K, MAPK, WNT, and NFkB. They also modulate the epigenetic machinery by regulating the expression and activity of DNA methyltransferases, histone modifiers and miRNA. This further enhances their transcriptional modulation of key cellular regulators. In this review, we comprehensively assess the impact of ITCs such as sulforaphane, phenethyl isothiocyanate, benzyl isothiocyanate and allyl isothiocyanate on cancer and document their effect on various molecular targets. Overall, this will facilitate consolidation of the current understanding of the anti-cancer and epigenetic modulatory potential of these compounds and recognize the gaps in literature. Further, we discuss avenues of future research to develop these compounds as potential therapeutic entities.
Sulfur-containing spice compounds possess diverse biological functions and play an important role in food, chemicals, pharmaceuticals, and agriculture. The development of functional spices has become increasingly popular, especially for medicinal functions for dietary health. Thus, this review focuses on the properties and functions of sulfur-containing spice compounds, including antioxidant, anti-inflammatory, antiobesity, anticancer, antibacterial, and insecticidal functions, among others. Developments over the last five years concerning the properties of sulfur-containing spice compounds are summarized and discussed.
BACKGROUND
Allyl isothiocyanate (AITC) is a volatile organic compound with a potent insecticidal activity to the stored‐grain pest Sitophilus zeamais Motschulsky, which severely damages grain storage and container transport worldwide. Our previous study showed that mitochondrial complex IV was the primary target of AITC in adult Sitophilus zeamais. To further verify the targets of AITC, we employed RNA interference (RNAi) by using double‐stranded RNA (dsRNA) to knockdown three core subunits of cytochrome c oxidase (COX)‐I, ‐II and ‐III in 18‐day‐old larvae prior to their exposure to AITC to detect susceptibility changes.
RESULTS
The susceptibility of dsRNACOX‐I and ‐II injection treatments to AITC significantly increased at 72 h while the mortality reached up to 85.56% and 67.78%, respectively, and dsRNACOX‐I and dsRNACOX‐II injection showed the same subcellular structural characteristics showing vacuolization and vague mitochondrial cristae and decrease of COX activity during AITC fumigation treatment, suggesting the potential of COX‐I and COX‐II as the targets of AITC. High mortality reached up to 75.55%, 71.88% and 82.22%, respectively, and the phenotype of larvae turning from milky white to dark brown in the thorax and death eventually was confirmed after dsRNACOX‐I, ‐II and ‐III injection.
CONCLUSION
COX‐I and ‐II were elucidated as the potential targets of AITC and dsRNACOX‐I, ‐II and ‐III have the potential to be developed into nucleic acid pesticides for their robust lethal effects and are worth pursuing for improving AITC fumigation activity in Sitophilus zeamais control.
Nature is a rich source of natural drug-like compounds with almost zero side effects. Phytochemicals better known as “Natural Products” are found abundantly in a number of plants. Since time immemorial, spices have been widely used in Indian cuisine as flavoring and colouring agents. Most of these spices and condiments are derived from various biodiversity hotspots in India (which contributes 75% of global spice production) and form the crux of India’s multidiverse and multicultural cuisine. Apart from their aroma, flavor and taste, these spices and condiments are known to possess several medicinal properties also. Most of these spices find considerable mention in Ayurveda, the indigenous system of medicine, as panaceas for several aliments. The antimicrobial, antioxidant, antiproliferative, antihypertensive and antidiabetic properties of several of these natural products are well documented in Ayurveda. These phytoconstituemts are known to act as functional immunoboosters, immunomodulators as well as anti-inflammatory agents. As anticancer agents, their mechanistic action involves cancer cell death via induction of apoptosis, necrosis and autophagy. The present review provides a comprehensive and collective update on the potential of 65 commonly used spices as well as their bioactive constituents as anticancer agents. The review also provides an in-depth update of all major in vitro, in vivo, clinical and pharmacological studies done on these spices with special emphasis on the potential of these spices and their bioactive constituents as potential functional foods for prevention, treatment and management of cancer.
Breast cancer metastasis is one of the major reasons for the high morbidity and mortality of breast cancer patients. In spite of surgical interventions, chemotherapy, radiation therapy and targeted therapy, some patients are considering alternative therapies with herbal/natural products. In the present study, we evaluated a well-characterized extract from the medicinal mushroom Ganoderma lucidum (GLE) for its affects on tumor growth and breast-to-lung cancer metastasis. MDA-MB-231 human breast cancer cells were implanted into the mammary fat pads of nude mice. GLE (100 mg/kg/every other day) was administered to the mice by an oral gavage for 4 weeks, and tumor size was measured using microcalipers. Lung metastases were evaluated by hematoxylin and eosin (H&E) staining. Gene expression in MDA-MB-231 cells was determined by DNA microarray analysis and confirmed by quantitative PCR. Identified genes were silenced by siRNA, and cell migration was determined in Boyden chambers and by wound-healing assay. Although an oral administration of GLE only slightly suppressed the growth of large tumors, the same treatment significantly inhibited the number of breast-to-lung cancer metastases. GLE also downregulated the expression of genes associated with invasive behavior (HRAS, VIL2, S100A4, MCAM, I2PP2A and FN1) in MDA-MB-231 cells. Gene silencing of HRAS, VIL2, S100A4, I2PP2A and FN1 by siRNA suppressed migration of MDA-MB‑231 cells. Our study suggests that an oral administration of GLE can inhibit breast-to-lung cancer metastases through the downregulation of genes responsible for cell invasiveness. The anti-metastatic benefits of GLE warrant further clinical studies.
Multidrug resistance-associated protein 1 (MRP1), a member of the ATP-binding cassette (ABC) superfamily of transporters, plays an important role in normal lung physiology by protecting cells against oxidative stress and toxic xenobiotics. The present study investigates the effects of allyl isothiocyanate (AITC) on MRP1 mRNA and MRP1 protein expression and transporter activity in the immortalised human bronchial epithelial cell line 16HBE14o-. MRP1 mRNA and MRP1 protein expression in 16HBE14o- cells that were treated with allyl isothiocyanate were analysed by real-time PCR assay and Western blotting. The transport of carboxyfluorescein, a known MRP1 substrate, was measured by functional flow cytometry to evaluate MRP1 activity. Treatment with AITC at concentrations of 5-40 μ M increased MRP1 protein levels in a concentration-dependent manner. AITC treatments at concentrations of 1-40 μ M caused concentration-dependent increases in MRP1 mRNA levels that were up to seven times greater than the levels found in control cells. Finally, AITC treatment at concentrations of 5-40 μ M significantly increased MRP1-dependent efflux in 16HBE14o- cells. These results suggest that AITC can increase the expression and activity of MRP1 in 16HBE14o- cells in a concentration-dependent manner. The upregulation of MRP1 activity and expression by AITC could produce therapeutic effects in the treatment of lung disease.
Diet in human health is no longer simple nutrition, but in light of recent research, especially nutrigenomics, it is linked via evolution and genetics to cell health status capable of modulating apoptosis, detoxification, and appropriate gene response. Nutritional deficiency and disease especially lack of vitamins and minerals is well known, but more recently, epidemiological studies suggest a role of fruits and vegetables, as well as essential fatty acids and even red wine (French paradox), in protection against disease. In the early 1990s, various research groups started considering the use of antioxidants (e.g., melatonin, resveratrol, green tea, lipoic acid) and metabolic compounds (e.g., nicotinamide, acetyl-L-carnitine, creatine, coenzyme Q10) as possible candidates in neuroprotection. They were of course considered on par with snake oil salesman (women) at the time. The positive actions of nutritional supplements, minerals, and plant extracts in disease prevention are now mainstream and commercial health claims being made are subject to regulation in most countries. Apart from efficacy and finding, the right dosages, the safety, and especially the level of purification and lack of contamination are all issues that are important as their use becomes widespread. From the mechanistic point of view, most of the time these substances replenish the body's deficiency and restore normal function. However, they also exert actions that are not sensu stricto nutritive and could be considered pharmacological especially that, at times, higher intake than recommended (RDA) is needed to see these effects. Free radicals and neuroinflammation processes underlie many neurodegenerative conditions, even Parkinson's disease and Alzheimer's disease. Curcumin, carotenoids, acetyl-L-carnitine, coenzyme Q10, vitamin D, and polyphenols and other nutraceuticals have the potential to target multiple pathways in these conditions. In summary, augmenting neuroprotective pathways using diet and finding new natural substances that can be more efficacious, i.e., induction of health-promoting genes and reduction of the expression of disease-promoting genes, could be incorporated into neuroprotective strategies of the future.
Many plant-derived isothiocyanates (ITCs), which occur in human diet, are potent cancer chemopreventive agents in animals. Among the anticarcinogenic mechanisms that have been revealed for ITCs is the inhibition of cell proliferation. We report that exposure of cancer cells to either allyl-ITC (AITC), benzyl-ITC (BITC), or phenethyl-ITC (PEITC) for only 3 h was long enough for the inhibition of cell growth, based on a comparison of IC50 values; regardless of the origin of cancer cells; and even in drug-resistant cells that overexpressed multidrug resistance associated protein-1 (MRP-1) or P-glycoprotein-1 (Pgp-1). In contrast, the inhibitory effect of another ITC, sulforaphane (SF), on these cells was highly time dependent. The finding that some ITCs could inhibit the proliferation of cancer cells in a largely time-independent manner is significant because ITCs that enter the human body are rapidly cleared through urinary excretion. Using human promyelocytic leukemia HL60/S as model cells, and focusing on AITC and BITC, we found that these ITCs modulated multiple cellular targets involved in proliferation, including the disruption of mitochondrial membrane potential, activation of multiple caspases, arrest of cell cycle progression, and induction of differentiation. Again, only a 3-h incubation of the cells with the ITCs was enough to exert their full effect on these targets. Taken together, our findings suggest that selected ITCs can rapidly initiate growth inhibition of cancer cells by simultaneously modulating multiple cellular targets, and their antiproliferative activity may be largely unaffected by their metabolism and disposition in vivo.
Sulforaphane (SUL), an isothiocyanate found in broccoli and other cruciferous vegetables, has been shown to induce phase II detoxification enzymes, inhibit chemically induced mammary tumors in rats, and more recently to induce cell cycle arrest and apoptosis in cancer cells of the colon. Here, we provide evidence that SUL also acts as a breast cancer anti-proliferative agent. The BALB/c mouse mammary carcinoma cell line F3II was treated with SUL at concentrations up to 15 microM and examined for markers of cell cycle arrest and apoptosis. Treatment of asynchronous F3II cells with 15 microM SUL resulted in G2/M cell cycle arrest, elevated p34cdc2 (cdc2) kinase activity, Bcl-2 down-regulation, evidence of caspase activation, and aggregation of condensed nuclear chromatin. Subsequent exposure of synchronized cells to 15 microM SUL resulted in elevated numbers of prophase/prometaphase mitotic figures, indicating cell cycle progression beyond G2 and arrest early within mitosis. Moreover, cells treated with 15 microM SUL displayed aberrant mitotic spindles, and higher doses of SUL inhibited tubulin polymerization in vitro. In addition, BALB/c mice injected s.c. with F3II cells and subsequently injected daily i.v. with SUL (15 nmol/day for 13 days) developed significantly smaller tumors (approximately 60% less in mass) than vehicle-treated controls. Western blot analysis of tumor proteins demonstrated significantly (P<0.05) reduced PCNA and elevated PARP fragmentation in samples from animals dosed with SUL. Taken together, these results indicate that SUL has mammary cancer suppressive actions both in cell culture and in the whole animal. Inhibition of mammary carcinogenesis appears in part to involve perturbation of mitotic microtubules and early M-phase block associated with cdc2 kinase activation, indicating that cells arrest prior to metaphase exit.
Recommendations for intake of fruits and vegetables, whole grains, lean meats, and low-fat dairy form the underpinning of dietary guidance for health promotion. We examined the association of a summary index of food consumption behaviors compatible with the spirit of prevailing dietary guidance and mortality. We used data from the NIH-American Association of Retired Persons cohort (n = 350,886), aged 50-71 y and disease free at baseline in 1995-1996, to examine the association of a dietary behavior score (DBS) with mortality after 10.5 y of follow-up (deaths, n = 29,838). The DBS included 6 equally weighted components derived from responses to questions on usual dietary behaviors related to consumption of fruits, vegetables, low-fat dairy, whole grains, lean meat and poultry, and discretionary fat. The covariate-adjusted association of DBS and mortality from all causes, cancer, and coronary heart disease was examined using Cox proportional hazards regression methods. Compared with those in the lowest one-fifth of DBS, the multivariate-adjusted relative risk of mortality in the highest one-fifth of the DBS was 0.75 (95% CI, 0.70-0.80) in women and 0.79 (95% CI, 0.75-0.83) in men (P-trend < 0.0001). The inverse association of DBS and mortality was significant in both genders in nearly all categories of covariates. Similar trends were observed for DBS associations with mortality from cancer and heart disease. Nearly 12% of the covariate-adjusted population risk of mortality was attributable to nonconformity with dietary recommendations. Adoption of recommended dietary behaviors was associated with lower mortality in both men and women independent of other lifestyle risk factors.
Urinary bladder cancer is the fourth most common malignancy in the Western world. Transitional cell carcinoma (TCC) is the
most common subtype, accounting for about 90% of all bladder cancers. The TP53 gene plays an essential role in the regulation of the cell cycle and apoptosis and therefore contributes to cellular transformation
and malignancy; however, little is known about the differential gene expression patterns in human tumors that present with
the wild-type or mutated TP53 gene. Therefore, because gene profiling can provide new insights into the molecular biology of bladder cancer, the present
study aimed to compare the molecular profiles of bladder cancer cell lines with different TP53 alleles, including the wild type (RT4) and two mutants (5637, with mutations in codons 280 and 72; and T24, a TP53 allele encoding an in-frame deletion of tyrosine 126). Unsupervised hierarchical clustering and gene networks were constructed
based on data generated by cDNA microarrays using mRNA from the three cell lines. Differentially expressed genes related to
the cell cycle, cell division, cell death, and cell proliferation were observed in the three cell lines. However, the cDNA
microarray data did not cluster cell lines based on their TP53 allele. The gene profiles of the RT4 cells were more similar to those of T24 than to those of the 5637 cells. While the deregulation
of both the cell cycle and the apoptotic pathways was particularly related to TCC, these alterations were not associated with
the TP53 status.
KeywordsGene expression–Gene network–
TP53
–Urinary bladder transitional cell carcinoma
To investigate the anti-tumor effect of capsaicin on human pharyngeal squamous carcinoma cells (FaDu).
The expression of apoptosis/cell cycle-related proteins (or genes) was examined by reverse transcriptase- polymerase chain reaction, western blotting and ELISA methods, while the apoptotic cell population, cell morphology and DNA fragmentation levels were assessed using flow cytometry, fluorescence microscopy and agarose gel electrophoresis.
Capsaicin was found to inhibit the growth and proliferation of FaDu cells in a dose- and time-dependent manner. Apoptotic cell death was confirmed by observing increases in nuclear condensation, nuclear DNA fragmentation and sub-G1 DNA content. The observed increase in cytosolic cytochrome c, activation of caspase 3 and PARP (p85) levels following capsaicin treatment indicated that the apoptotic response was mitochondrial pathway-dependent. Gene/protein expression analysis of Bcl-2, Bad and Bax further revealed decreased anti-apoptotic Bcl-2 protein and increased pro-apoptotic Bad/Bax expression. Furthermore, capsaicin suppressed the cell cycle progression at the G1/S phase in FaDu cells by decreasing the expression of the regulators of cyclin B1 and D1, as well as cyclin-dependent protein kinases cdk-1, cdk-2 and cdk-4.
Our current data show that capsaicin induces apoptosis in FaDu cells and this response is associated with mitochondrial pathways, possibly by mediating cell cycle arrest at G1/S.
Allyl isothiocyanate (AITC) occurs in many commonly consumed cruciferous vegetables and exhibits significant anti-cancer activities. Available data suggest that it is particularly promising for bladder cancer prevention and/or treatment. Here, we show that AITC arrests human bladder cancer cells in mitosis and also induces apoptosis. Mitotic arrest by AITC was associated with increased ubiquitination and degradation of α- and β-tubulin. AITC directly binds to multiple cysteine residues of the tubulins. AITC induced mitochondrion-mediated apoptosis, as shown by cytochrome c release from mitochondria to cytoplasm, activation of caspase-9 and caspase-3, and formation of TUNEL-positive cells. Inhibition of caspase-9 blocked AITC-induced apoptosis. Moreover, we found that apoptosis induction by AITC depended entirely on mitotic arrest and was mediated via Bcl-2 phosphorylation at Ser-70. Pre-arresting cells in G(1) phase by hydroxyurea abrogated both AITC-induced mitotic arrest and Bcl-2 phosphorylation. Overexpression of a Bcl-2 mutant prevented AITC from inducing apoptosis. We further showed that AITC-induced Bcl-2 phosphorylation was caused by c-Jun N-terminal kinase (JNK), and AITC activates JNK. Taken together, this study has revealed a novel anticancer mechanism of a phytochemical that is commonly present in human diet.
S100P was recently found to be overexpressed in a variety of cancers and is considered a potential target for cancer therapy, but the functional role or mechanism of action of S100P in colon cancer is not fully understood. In the present study, we knocked down the gene expression of S100P in colon cancer cells using lentivirus-mediated RNA interference. This step resulted in significant inhibition of cancer cell growth, migration and invasion in vitro and tumor growth and liver metastasis in vivo. Moreover, S100P downstream target proteins were identified by proteomic analysis in colon cancer DLD-1 cells with deletion of S100P. Knockdown of S100P led to downregulation of thioredoxin 1 and β-tubulin and upregulation of Rho guanosine diphosphate (GDP) dissociation inhibitor α (RhoGDIA), all potential therapeutic targets in cancer. Taken together, these findings suggest that S100P plays an important role in colon tumorigenesis and metastasis, and the comprehensive and comparative analyses of proteins associated with S100P could contribute to understanding the downstream signal cascade of S100P, leading to tumorigenesis and metastasis.
Effective clinical management of prostate cancer (PCA) has been challenged by significant intratumoural heterogeneity on the genomic and pathological levels and limited understanding of the genetic elements governing disease progression. Here, we exploited the experimental merits of the mouse to test the hypothesis that pathways constraining progression might be activated in indolent Pten-null mouse prostate tumours and that inactivation of such progression barriers in mice would engender a metastasis-prone condition. Comparative transcriptomic and canonical pathway analyses, followed by biochemical confirmation, of normal prostate epithelium versus poorly progressive Pten-null prostate cancers revealed robust activation of the TGFβ/BMP-SMAD4 signalling axis. The functional relevance of SMAD4 was further supported by emergence of invasive, metastatic and lethal prostate cancers with 100% penetrance upon genetic deletion of Smad4 in the Pten-null mouse prostate. Pathological and molecular analysis as well as transcriptomic knowledge-based pathway profiling of emerging tumours identified cell proliferation and invasion as two cardinal tumour biological features in the metastatic Smad4/Pten-null PCA model. Follow-on pathological and functional assessment confirmed cyclin D1 and SPP1 as key mediators of these biological processes, which together with PTEN and SMAD4, form a four-gene signature that is prognostic of prostate-specific antigen (PSA) biochemical recurrence and lethal metastasis in human PCA. This model-informed progression analysis, together with genetic, functional and translational studies, establishes SMAD4 as a key regulator of PCA progression in mice and humans.
The standard palliative chemotherapy for pancreatic ductal adenocarcinoma (PDAC) is gemcitabine-based chemotherapy; however, PDAC still presents a major therapeutic challenge. The aims of this study were to investigate the expression pattern of genes involved in gemcitabine sensitivity in resected PDAC tissues and to determine correlations of gene expression with treatment outcome.
We obtained formalin-fixed paraffin-embedded (FFPE) tissue samples from 70 patients with PDAC. Of the 70 patients, 40 received gemcitabine-based adjuvant chemotherapy (AC). We measured hENT1, dCK, CDA, RRM1, and RRM2 messenger RNA (mRNA) levels by quantitative real-time reverse transcription-polymerase chain reaction and determined the combined score (GEM score), based on the expression levels of hENT1, dCK, RRM1, and RRM2, to investigate the association with survival time. By determining the expression levels of these genes in endoscopic ultrasound-guided fine needle aspiration (EUS-FNA) cytologic specimens, we investigated the feasibility of individualized chemotherapy.
High dCK (P = .0067), low RRM2 (P = .003), and high GEM score (P = .0003) groups had a significantly longer disease-free survival in the gemcitabine-treated group. A low GEM score (<2) was an independent predictive marker for poor outcome to gemcitabine-based AC as shown by multivariate analysis (P = .0081). Altered expression levels of these genes were distinguishable in microdissected neoplastic cells from EUS-FNA cytologic specimens.
Quantitative analyses of hENT1, dCK, RRM1, and RRM2 mRNA levels using FFPE tissue samples and microdissected neoplastic cells from EUS-FNA cytologic specimens may be useful in predicting the gemcitabine sensitivity of patients with PDAC.
Isothiocyanates (ITCs) are present as glucosinolates in various cruciferous vegetables. Allyl isothiocyanate (AITC) is one of the common naturally occurring isothiocyanates. Recent studies have shown that AITC significantly inhibited survival of leukemia HL-60, bladder cancer UM-UC-3 and colon cancer HT-29 cells in vitro. In this study, we demonstrate that AITC significantly decreased proliferation and viability of human brain malignant glioma GBM 8401 cells in a dose-dependent manner with IC50 9.25+/-0.69 microM for 24 h-treatment. The analysis of cell cycle distribution also showed that AITC induced significantly G2/M arrest and sub-G1 phase (apoptotic population) in GBM 8401 cells. AITC markedly reduced the CDK1/cyclin B activity and protein levels by CDK1 activity assay and Western blot analysis. AITC-induced apoptotic cell death and this evidence was confirmed by morphological assessment and DAPI staining. Pretreatment with specific inhibitors of caspase-3 (Z-DEVE-FMK) and -9 (Z-LEHD-FMK) significantly reduced caspase-3 and -9 activity in GBM 8401 cells. Western blot analysis and colorimetric assays also displayed that AITC caused a time-dependent increase in cytosolic cytochrome c, pro-caspase-9, Apaf-1, AIF, Endo G and the stimulated caspase-9 and -3 activity. Our results suggest that AITC is a potent anti-human brain malignant glioma drug and it shows a remarkable action on cell cycle arrest before commitment for apoptosis is reached.
Intake of cruciferous vegetables, a rich source of dietary isothiocyanates, has been inversely associated with risk of bladder cancer. Due to the potent antiproliferative effects of dietary isothiocyanates on bladder cancer in in vitro and in vivo models, cruciferous vegetable intake may also play a role in survival among patients with bladder cancer.
Using information obtained from the Roswell Park Cancer Institute Tumor Registry, patient medical records, and routinely collected questionnaire data, we examined potential associations between intake of cruciferous vegetables and survival among bladder cancer patients. As cooking can substantially reduce or destroy isothiocyanates, consumption of raw versus cooked cruciferous vegetables was examined separately. Hazard ratios (HR) and 95% confidence intervals (CI) were estimated using Cox proportional hazard models.
A total of 239 bladder cancer patients were included in the study. After an average of 8 years of follow-up, 179 deaths occurred, with 101 deaths attributable to bladder cancer. After adjustment for other prognostic factors, a strong and significant inverse association was observed between bladder cancer mortality and broccoli intake, in particular raw broccoli intake (> or =1 versus <1 serving per month; HR for overall death, 0.57; 95% CI, 0.39-0.83; HR for disease-specific death, 0.43; 95% CI, 0.25-0.74). There were no significant associations for total vegetables, total fruits, or other individual cruciferous vegetables.
Considering the strong preclinical evidence, intake of broccoli may improve bladder cancer survival.
Further prospective investigation is warranted to confirm the potential role of cruciferous vegetables in bladder cancer prognosis.
Bladder cancer is one of the common human cancers and also has a very high recurrence rate. There is a great need for agents capable of inhibiting bladder cancer development and recurrence. Here, we report that allyl isothiocyanate (AITC), an ingredient of many common cruciferous vegetables, potently inhibited the proliferation of bladder carcinoma cell lines in vitro [half maximal inhibitory concentration (IC(50)) of 2.7-3.3 microM], which was associated with profound G(2)/M arrest and apoptosis. In contrast, AITC was markedly less toxic to normal human bladder epithelial cells (IC(50) of 69.4 microM). AITC was then evaluated in two rat bladder cancer models in vivo (an orthotopic model and a subcutaneous model). The orthotopic model closely mimics human bladder cancer development and recurrence. We show that a low oral dose of AITC (1 mg/kg) significantly inhibited the development and muscle invasion of the orthotopic bladder cancers but was ineffective against the subcutaneous xenografts of the same cancer cells in the same animals. This differential effect was explained by our finding that urinary levels of AITC equivalent were two to three orders of magnitude higher than that in the plasma and that its levels in the orthotopic cancer tissues were also three orders of magnitude higher than that in the subcutaneous cancer tissues. Moreover, we show that AITC is a multi-targeted agent against bladder cancer. In conclusion, AITC is selectively delivered to bladder cancer tissue through urinary excretion and potently inhibits bladder cancer development and invasion.
Inherent cancer phenotypes that are independent of fluctuating cross-talk with the surrounding tissue matrix are highly desirable candidates for targeting tumor cells. Our novel study design uses epithelial cell lines derived from low versus high histologic grade primary breast cancer to effectively diminish the breadth of transient variability generated within the tumor microenvironment of the host, revealing a "paracrine-independent expression of grade-associated" (PEGA) gene signature. PEGA members extended beyond "proliferation-driven" signatures commonly associated with aggressive, high-grade breast cancer. The calcium-binding protein S100P was prominent among PEGA genes overexpressed in high-grade tumors. A three-member fingerprint of S100P-correlated genes, consisting of GPRC5A, FXYD3, and PYCARD, conferred poor outcome in multiple breast cancer data sets, irrespective of estrogen receptor status but dependent on tumor size (P < 0.01). S100P silencing markedly diminished coregulated gene transcripts and reversed aggressive tumor behavior. Exposure to pathway-implicated agents, including the calmodulin inhibitor N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide, phenothiazine, and chlorpromazine, resulted in rapid apoptotic cell death in high-grade tumor cells resistant to the chemotherapeutic drug cisplatin. This is the first comprehensive study describing molecular phenotypes intimately associated with histologic grade whose expression remains relatively fixed despite an unavoidably changing environment to which tumor cells are invariably exposed.
The authors investigate the antiangiogenic and proapoptotic effects of mustard essential oil containing allyl isothiocyanate (AITC) and explore its mechanism of action on Ehrlich ascites tumor (EAT) cells. Swiss albino mice transplanted with EAT cells were used to study the effect of AITC. AITC was effective at a concentration of 10 mum as demonstrated by the inhibition of proliferation of EAT cells when compared with the normal HEK293 cells. It significantly reduced ascites secretion and tumor cell proliferation by about 80% and inhibited vascular endothelial growth factor expression in tumor-bearing mice in vivo. It also reduced vessel sprouting and exhibited potent antiangiogenic activity in the chorioallantoic membrane and cornea of the rat. AITC arrested the growth of EAT cells by inducing apoptosis and effectively arrested cell cycle progression at the G1 phase. The results clearly suggest that AITC inhibits tumor growth by both antiangiogenic and proapoptotic mechanisms.
The mouse Nedd5 gene encodes a 41.5-kD GTPase similar to the Saccharomyces and Drosophila septins essential for cytokinesis. Nedd5 accumulates near the contractile ring from anaphase through telophase, and finally condenses into the midbody. Microinjection of anti-Nedd5 antibody interferes with cytokinesis, giving rise to binucleated cells. In interphase and postmitotic cells, Nedd5 localizes to fibrous or granular structures depending on the growth state of the cell. The Nedd5-containing fibers are disrupted by microinjection of GTPgammaS and by Nedd5 mutants lacking GTP-binding activity, implying that GTP hydrolysis is required for its assembly. The Nedd5-containing fibers also appear to physically contact actin bundles and focal adhesion complexes and are disrupted by cytochalasin D, C3 exoenzyme, and serum starvation, suggesting a functional interaction with the actin-based cytoskeletal systems in interphase cells.
Cell death and differentiation is a monthly research journal focused on the exciting field of programmed cell death and apoptosis. It provides a single accessible source of information for both scientists and clinicians, keeping them up-to-date with advances in the field. It encompasses programmed cell death, cell death induced by toxic agents, differentiation and the interrelation of these with cell proliferation.
Although several genes that might mediate p53-induced apoptosis have been proposed, none have previously been shown to play an essential role in this process through a rigorous gene disruption approach. We used a gene-targeting approach to evaluate p53-mediated death in human colorectal cancer cells. Expression of p53 in these cells induces growth arrest through transcriptional activation of the cyclin-dependent kinase inhibitor p21. If p21 is disrupted via gene targeting, the cells die through apoptosis. If the PUMA gene is also disrupted in such cells, apoptosis is prevented. The effects of PUMA on apoptosis were observed after exogenous overexpression of p53 as well as after exposure to hypoxia, a physiologic activator of p53, and DNA damage. The PUMA protein interacts with Bcl-X(L) and promotes mitochondrial translocation and multimerization of Bax. Accordingly, genetic disruption of BAX makes cells resistant to the apoptosis resulting from PUMA expression. These results suggest that the balance between PUMA and p21 is pivotal in determining the responses to p53 activation and provide a model for understanding the basis of p53 mutations in human cancer.
Smoking is a major risk factor for urothelial cell carcinoma of the bladder (UCC). Mutations in the FGFR3 and TP53 genes have been shown to define two distinct pathways in superficial papillary and invasive UCC disease, respectively. We investigated the relationship between smoking and these mutations by means of denaturing high performance liquid chromatography and sequencing for 110 primary UCC of the bladder. This study included 48 current smokers, 31 ex-smokers and 31 non-smokers. Thirty-five of the tumors were stage pTa, 40 pT1 and 35 > or =pT2. Fourteen of the tumors were grade 1, 37 were grade 2 and 59 grade 3. Smoking was associated with high stage (P = 0.03) and high grade tumors (P = 0.006). Twenty-two of the 110 tumors studied harbored TP53 mutations (20%) and 43 harbored FGFR3 mutations (39%). Odds ratios (OR) were higher for TP53 mutations in current smokers [OR, 2.25; 95% confidence interval (95% CI), 0.65-7.75] and ex-smokers (OR, 1.62; 95% CI, 0.41-6.42) than in non-smokers. Double TP53 mutations and the A:T-->G:C TP53 mutation pattern was found only in current smokers. Patients with the FGFR3(wild-type)/TP53(mutated) genotype had significantly higher levels of tobacco consumption, as measured in pack-years (P = 0.01). Smoking influenced neither the frequency nor the pattern of FGFR3 mutations. Our results suggest that smoking is associated with invasive and high grade UCCs, at initial presentation, and influenced TP53 or the molecular pathway defined by these mutations. In contrast, FGFR3 mutations are not affected by smoking and probably result from endogenous alterations. These data have potential implications for clinical management and prevention strategies.
The clinical course of renal cell carcinoma (RCC) can be difficult to predict. In this study we evaluated the prognostic value of anillin and Ki-67 in predicting survival in RCC. Immunohistochemical analysis using anillin and Ki-67 antibodies was performed on tissue microarrays constructed from paraffin-embedded specimens from 152 patients with primary RCC. The mean follow-up time was 90 months. Levels of anillin and Ki-67 staining were correlated with clinical factors, pathological features and survival. Anillin expression in cytoplasmic and nuclear fractions of the RCC cell line 786-O was examined using Western blot analysis. Cytoplasmic anillin immunopositivity was detected in 121 (83%) tumours. Nuclear anillin expression was present in 40 (27%) tumours and increased Ki-67 activity in 98 (66%) tumours. A positive association was found between nuclear anillin and Ki-67 proliferation activity (p=0.005). The mean RCC-specific survival times for anillin immunopositive and immunonegative tumours were 158 (95% CI 143-173) and 109 (78-141) months, respectively, with p=0.03. Increased Ki-67 activity showed a tendency towards a poorer prognosis, although this was not statistically significant. In the Cox regression analysis for cytoplasmic anillin, nuclear anillin or Ki-67 rate, and age, gender, stage and nuclear grade, the only significant factor in RCC-specific survival was stage (p<0.001). Western blot analysis showed anillin expression in both nuclear and cytosolic fractions of the RCC cell line. To conclude, anillin expression can be observed both in the cytoplasm and nuclei in patients with RCC. Cytoplasmic anillin expression is a marker of favourable prognosis in RCC patients.
Allyl isothiocyanate (AITC) is present in plants of the cruciferous family and is abundant in mustard seed. Due to its high bioavailability in urine after ingestion, AITC has been considered a promising antineoplastic agent against bladder cancer. Because TP53 mutations are the most common alterations in bladder cancer cells and are frequently detected in in situ carcinomas, in this study, we investigated whether the AITC effects in bladder cancer cells are dependent on the TP53 status. Two bladder transitional carcinoma cell lines were used: RT4, with wild-type TP53; and T24, mutated TP53 gene. AITC was tested at concentrations of 0.005, 0.0625, 0.0725, 0.0825, 0.0925, 0.125 and 0.25 μM in cytotoxicity, cell and clonogenic survival assays, comet and micronucleus assays and for its effects on cell cycle and apoptosis by flow cytometry and on TP53 gene expression. The data showed increased primary DNA damage in both cell lines; however, lower concentrations of AITC were able to induce genotoxicity in the mutant cells for the TP53 gene. Furthermore, the results demonstrated increased apoptosis and necrosis rates in the wild-type cells, but not in mutated TP53 cells, and cell cycle arrest in the G2 phase for mutated cells after AITC treatment. No significant differences were detected in TP53 gene expression in the two cell lines. In conclusion, AITC caused cell cycle arrest, increased apoptosis rates and varying genotoxicity dependent on the TP53 status. However, we cannot rule out the possibility that those differences could reflect other intrinsic genetic alterations in the examined cell lines, which may also carry mutations in genes other than TP53. Therefore, further studies using other molecular targets need to be performed to better understand the mechanisms by which AITC may exert its antineoplastic properties against tumor cells.
Cancer is a multistep process resulting in uncontrolled cell division. It results from aberrant signaling pathways that lead to uninhibited cell division and growth. Various recent epidemiological studies have indicated that consumption of cruciferous vegetables, such as garden cress, broccoli, etc., reduces the risk of cancer. Isothiocyanates (ITCs) have been identified as major active constituents of cruciferous vegetables. ITCs occur in plants as glucosinolate and can readily be derived by hydrolysis. Numerous mechanistic studies have demonstrated the anticancer effects of ITCs in various cancer types. ITCs suppress tumor growth by generating reactive oxygen species or by inducing cycle arrest leading to apoptosis. Based on the exciting outcomes of preclinical studies, few ITCs have advanced to the clinical phase. Available data from preclinical as well as available clinical studies suggest ITCs to be one of the promising anticancer agents available from natural sources. This is an up-to-date exhaustive review on the preventive and therapeutic effects of ITCs in cancer.
Medicinal herbs, used in indigenous medicines in crude forms for the management of diabetes mellitus, contain both the organic and inorganic constituents. The aim of the study was to find out the hypoglycemic effect of Ficus racemosa in a group of diabetic subjects taking oral hypoglycemic drug. Twenty five of each, male and female, diabetic patients, selected from Fatima Jinnah Medical College, Lahore, Pakistan, taking oral hypoglycemic drug were included in this study and were given orally the extract (5 mL) of bark of Ficus racemosa (about 100 mg) two times for 15 days. Blood samples for estimation of blood glucose and parameters of liver and renal functions were estimated. It was observed that after taking the herb in combination with drug, blood glucose level (fasting and after breakfast) was markedly decreased in both male and female but significant difference was only observed in sugar level of males after 1.5 h after breakfast. To rule out herb toxicity, liver and renal functions tests of patients was also performed which were observed to be in normal range. Present investigation established a pharmacological evidence to support the folklore claim that Ficus racemosa is good anti-diabetic agent.
The two most commonly used methods to analyze data from real-time, quantitative PCR experiments are absolute quantification and relative quantification. Absolute quantification determines the input copy number, usually by relating the PCR signal to a standard curve. Relative quantification relates the PCR signal of the target transcript in a treatment group to that of another sample such as an untreated control. The 2(-DeltaDeltaCr) method is a convenient way to analyze the relative changes in gene expression from real-time quantitative PCR experiments. The purpose of this report is to present the derivation, assumptions, and applications of the 2(-DeltaDeltaCr) method. In addition, we present the derivation and applications of two variations of the 2(-DeltaDeltaCr) method that may be useful in the analysis of real-time, quantitative PCR data. (C) 2001 Elsevier science.
Malignant glioma, also known as brain cancer, is the most common intracranial tumor, having an extremely high mortality and recurrence rate. The survival rate of the affected patients is very low and treatment is difficult. Hence, growth inhibition of glioma has become a hot topic in the study of brain cancer treatment. Among the various isothiocyanate compounds, it has been confirmed that benzyl isothiocyanate (BITC) can inhibit the growth of a variety of tumors, including leukemia, glioma and lung cancer, both inside and outside the body. This study explored inhibitory effects of BITC on human glioma U87MG cells, as well as potential mechanisms. It was found that BITC could inhibit proliferation, induce apoptosis and arrest cell cycling of U87MG cells. In addition, it inhibited the expression of SOD and GSH, and caused oxidative stress to tumor cells. Therefore, it is believed that BITC can inhibit the growth of U87MG cells outside the body. Its mechanism may be related to the fact that BITC can cause oxidative stress to tumor cells.
Natural products including botanicals for both therapy of clinical manifestations of atherosclerosis and reduction of atherosclerosis risk factors are topics of recent patents. Only a few recent patents are relevant to the direct anti-atherosclerotic therapy leading to regression of atherosclerotic lesions. Earlier, using a cellular model we have developed and patented several anti-atherosclerotic drugs. The AMAR (Atherosclerosis Monitoring and Atherogenicity Reduction) study was designed to estimate the effect of two-year treatment with time-released garlic-based drug Allicor on the progression of carotid atherosclerosis in 196 asymptomatic men aged 40-74 in double-blinded placebo-controlled randomized clinical study. The primary outcome was the rate of atherosclerosis progression, measured by high-resolution B-mode ultrasonography as the increase in carotid intima-media thickness (IMT) of the far wall of common carotid arteries. The mean rate of IMT changes in Allicor-treated group (-0.022±0.007 mm per year) was significantly different (P=0.002) from the placebo group in which there was a moderate progression of 0.015±0.008 mm at the overall mean baseline IMT of 0.931±0.009 mm. A significant correlation was found between the changes in blood serum atherogenicity (the ability of serum to induce cholesterol accumulation in cultured cells) during the study and the changes in intima-media thickness of common carotid arteries (r=0.144, P=0.045). Thus, the results of AMAR study demonstrate that long-term treatment with Allicor has a direct anti-atherosclerotic effect on carotid atherosclerosis and this effect is likely to be due to serum atherogenicity inhibition. The beneficial effects of other botanicals including Inflaminat (calendula, elder and violet), phytoestrogen-rich Karinat (garlic powder, extract of grape seeds, green tea leafs, hop cones, β-carotene, α-tocopherol and ascorbic acid) on atherosclerosis have also been revealed in clinical studies which enforces a view that botanicals might represent promising drugs for anti-atherosclerotic therapy.
S100 is a multigenic family of non-ubiquitous Ca2+-modulated proteins of the EF-hand type expressed in vertebrates exclusively and implicated in intracellular and extracellular regulatory activities. Within cells, most of S100 members exist in the form of antiparallely packed homodimers (in some cases heterodimers), capable of functionally crossbridging two homologous or heterologous target proteins in a Ca2+-dependent (and, in some instances, Ca2+-independent) manner. S100 oligomers can also form, under the non-reducing conditions found in the extracellular space and/or within cells upon changes in the cell redox status. Within cells, S100 proteins have been implicated in the regulation of protein phosphorylation, some enzyme activities, the dynamics of cytoskeleton components, transcription factors, Ca2+ homeostasis, and cell proliferation and differentiation. Certain S100 members are released into the extracellular space by an unknown mechanism. Extracellular S100 proteins stimulate neuronal survival and/or differentiation and astrocyte proliferation, cause neuronal death via apoptosis, and stimulate (in some cases) or inhibit (in other cases) the activity of inflammatory cells. A cell surface receptor, RAGE, has been identified on inflammatory cells and neurons for S100A12 and S100B, which transduces S100A12 and S100B effects. It is not known whether RAGE is a universal S100 receptor, S100 members interact with other cell surface receptors, or S100 protein interaction with other extracellular factors specifies the biological effects of a given S100 protein on a target cell. The variety of intracellular target proteins of S100 proteins and, in some cases, of a single S100 protein, and the cell specificity of expression of certain S100 members suggest that these proteins might have a role in the fine regulation of effector proteins and/or specific steps of signaling pathways/cellular functions. Future analyses should discriminate between functionally relevant S100 interactions with target proteins and in vitro observations devoid of physiological importance.
Urothelial carcinoma of the urinary bladder (UCB) is a highly heterogeneous malignancy that causes significant morbidity and mortality. Despite advances in surgical and medical treatment, there has been no change in mortality in UCB over the past decades. Standard pathological features (stage, grade, nodal status) provide only limited information regarding biological potential and clinical behavior. Molecular biomarkers may shed light on important mechanisms of pathogenesis, provide useful additional prognostic information, and serve as targets for therapy. This review summarizes recent advances and the most promising UCB tissue and blood biomarkers of the past few years. We discuss the predictive and prognostic value of biomarkers at different stages of UCB. There is no doubt that a panel of biomarkers will eventually improve our clinical decision-making with regard to treatment and follow-up.
Naturally occurring allyl isothiocyanate (AITC) was recently shown to be selectively delivered to bladder cancer tissue via
urinary excretion and to inhibit bladder cancer growth and muscle invasion in an animal model. AITC is excreted in urine mainly
as N-acetyl-S-(N-allylthiocarbamoyl)cysteine, more commonly known as the N-acetylcysteine conjugate (NAC-AITC). We show here that treatment of human bladder cancer UM-UC-3 cells or rat bladder cancer
AY-27 cells with NAC-AITC at 15 μM results in significant inhibition of cell growth and proliferation, together with cell
cycle arrest and apoptosis. We also show that NAC-AITC administered orally at 10 μmol/kg body wt inhibits cancer growth by
40% and muscle invasion by 49% in an orthotopic rat bladder cancer model. Furthermore, the anticancer activity of NAC-AITC
is associated with the modulation of several important molecular targets, including downregulation of both α-tubulin and β-tubulin,
activation of caspase-3 and downregulation of vascular endothelial growth factor. These results are similar to those shown
previously for AITC and are consistent with the understanding that NAC-AITC is a carrier of AITC. Furthermore, comparison
of the pharmacokinetic and physical properties of NAC-AITC with those of AITC suggests that NAC-AITC is superior to AITC for
potential use for prevention and therapy of bladder cancer.
S100P is a 95-amino-acid protein and a member of the S100 family. It was first purified from placenta. The promoter area of S100P has binding sites for SMAD, STAT/CREB and SP/KLF, key regulatory elements participating in transcriptional activation of the S100P gene. Increased levels of S100P have been observed in multiple tumor cell lines and breast, pancreas, lung and ovary carcinomas. S100P has been shown to mediate tumor growth, metastasis and invasion through the binding of Ca(2+) ions, receptor for advanced glycation end products, cytoskeletal protein ezrin, calcyclin-binding protein/Siah-1-interacting protein and cathepsin D. S100P could potentially serve as diagnostic marker, prognostic/predictive indicator and therapy target for different carcinomas.
The clinical course of renal cell carcinoma (RCC) can be difficult to predict. In this study we evaluated the prognostic value of anillin and Ki-67 in predicting survival in RCC. Immunohistochemical analysis using anillin and Ki-67 antibodies was performed on tissue microarrays constructed from paraffin-embedded specimens from 152 patients with primary RCC. The mean follow-up time was 90 months. Levels of anillin and Ki-67 staining were correlated with clinical factors, pathological features and survival. Anillin expression in cytoplasmic and nuclear fractions of the RCC cell line 786-O was examined using Western blot analysis. Cytoplasmic anillin immunopositivity was detected in 121 (83%) tumours. Nuclear anillin expression was present in 40 (27%) tumours and increased Ki-67 activity in 98 (66%) tumours. A positive association was found between nuclear anillin and Ki-67 proliferation activity (p=0.005). The mean RCC-specific survival times for anillin immunopositive and immunonegative tumours were 158 (95% CI 143-173) and 109 (78-141) months, respectively, with p=0.03. Increased Ki-67 activity showed a tendency towards a poorer prognosis, although this was not statistically significant. In the Cox regression analysis for cytoplasmic anillin, nuclear anillin or Ki-67 rate, and age, gender, stage and nuclear grade, the only significant factor in RCC-specific survival was stage (p<0.001). Western blot analysis showed anillin expression in both nuclear and cytosolic fractions of the RCC cell line. To conclude, anillin expression can be observed both in the cytoplasm and nuclei in patients with RCC. Cytoplasmic anillin expression is a marker of favourable prognosis in RCC patients.
Transitional cell (urothelial) carcinoma of the bladder is the second most common urologic malignancy and is one of the best understood neoplasms, with relatively well-defined pathogenetic pathways, natural history, and tumor biology. Conventional clinical and pathologic parameters are widely used to grade and stage tumors and to predict clinical outcome of transitional cell carcinoma; but the predictive ability of these parameters is limited, and there is a lack of indices that could allow prospective assessment of risk for individual patients. In the last decade, a wide range of candidate biomarkers representing key pathways in carcinogenesis have been reported to be clinically relevant and potentially useful as diagnostic and prognostic molecular markers, and as potential therapeutic targets. The use of molecular markers has facilitated the development of novel and more accurate diagnostic, prognostic, and therapeutic strategies. FGFR3 and TP53 mutations have been recognized as key genetic pathways in the carcinogenesis of transitional cell carcinoma. FGFR3 appears to be the most frequently mutated oncogene in transitional cell carcinoma; its mutation is strongly associated with low tumor grade, early stage, and low recurrence rate, which confer a better overall prognosis. In contrast, TP53 mutations are associated with higher tumor grade, more advanced stage, and more frequent tumor recurrences. These molecular markers offer the potential to characterize individual urothelial neoplasms more completely than is possible by histologic evaluation alone. Areas in which molecular markers may prove valuable include prediction of tumor recurrence, molecular staging of transitional cell carcinoma, detection of lymph node metastasis and circulating cancer cells, identification of therapeutic targets, and prediction of response to therapy. With accumulating molecular knowledge of transitional cell carcinoma, we are closer to the goal of bridging the gap between molecular findings and clinical outcomes. Assessment of key genetic pathways and expression profiles could ultimately establish a set of molecular markers to predict the biological nature of tumors and to establish new standards for molecular tumor grading, classification, and prognostication. The main focus of this review is to discuss clinically relevant biomarkers that might be useful in the management of transitional cell carcinoma and to provide approaches in the analysis of molecular pathways that influence the clinical course of bladder cancer.
Protein-protein interactions between the Bcl2 family proteins regulate apoptosis. An imbalance of this interaction network due to the upregulation of the proto-oncogene Bcl2 leads to a resistance to apoptosis associated with tumor formation. Bcl2 overexpression inhibits BAX oligomerization and mitochondrial outer membrane (MOM) permeabilization. However, Bcl2 effects on earlier steps of BAX-mediated apoptosis are not fully understood. Bcl2 overexpression inhibits BAX insertion into the MOM but spontaneously increases BAX relocalization to the mitochondria. Also, a physical interaction between BAX and Bcl2 is necessary for these two effects to occur. Taken together, these results suggest upregulated Bcl2 stabilizes BAX loose binding to mitochondrial membranes, inhibiting its insertion into the MOM and consequently cytochrome c release.
Bladder cancer is the fifth most common human malignancy and the second most frequently diagnosed genitourinary tumor after prostate cancer. The majority of malignant tumors arising in the urinary bladder are urothelial carcinomas. Clinically, superficial bladder tumors (stages Ta and Tis) account for 75% to 85% of neoplasms, while the remaining 15% to 25% are invasive (T1, T2–T4) or metastatic lesions at the time of initial presentation. Several studies have revealed that distinct genotypic and phenotypic patterns are associated with early vs. late stages of bladder cancer. Early superficial disease appears to segregate into 2 main pathways: (1) superficial papillary bladder tumors, which are characterized by gain-of-function mutations affecting oncogenes such as H-RAS, FGFR3, and PI3K, and deletions of the long arm of chromosome 9 (9q); (2) Carcinoma in situ, a “flat” high grade lesion considered to be a precursor of invasive cancer, is characterized by loss-of-function mutations affecting tumor suppressor genes, such as p53, RB, and PTEN. Based on these data, a model for bladder tumor progression has been proposed in which 2 separate genetic pathways characterize the evolution of early bladder neoplasms. Several molecular markers have been correlated with tumor stage, but the rationale for these 2 well-defined genetic pathways still remains unclear.
This review surveys some of the areas in which nutrients have been shown to have an impact on specific immune functions, the use of molecular and genetic tools to study molecular responses to dietary factors,and the metabolic consequences of food. It also explores the relationships between nutrient molecules, genetic polymorphisms, and the biological system as a whole, while providing a short introduction to nutrition immunology, nutrient-gene interactions and the novel technologies employed in nutrigenomics and nutrigenetics research.
Allyl isothiocyanate (AITC) is a constituent of cruciferous vegetables. It occurs widely in the human diet as a natural ingredient or food additive. AITC possesses numerous biochemical and physiological activities. It is cytotoxic and tumorigenic at high doses and also is a modulator of enzymes involved in metabolism of xenobiotics, including carcinogens. It is plausible that the wide consumption of dietary AITC may have profound effects on human health. To facilitate investigations of the effects of dietary AITC in humans, a method of measuring its uptake is needed. In this study, a urinary marker was developed for quantifying AITC uptake in humans. Four adult volunteers were asked to eat a meal containing brown mustard as the source of AITC. The 48-h urine samples were collected from these individuals and analyzed by reverse phase high performance liquid chromatography. A major urinary metabolite was found, which was identified as N-acetyl-S-(N-allylthiocarbamoyl)-L-cysteine, the N-acetylcysteine conjugate of AITC, by comparing its retention time and UV, nuclear magnetic resonance, and mass spectra with those of the synthetic standard. After ingestion of mustard, the AITC conjugate was detected in urine collected from 0 to 12 h. No conjugate was found in urine samples collected after 12 h. The major portion of this metabolite was excreted within 8 h. The average total excretion of AITC conjugate was 5.4 +/- 1.7 (SD) mg after consumption of 10 g of mustard and 12.8 +/- 2.0 mg when 20 g of mustard was consumed. Thus, a dose-dependent excretion of this metabolite was demonstrated.(ABSTRACT TRUNCATED AT 250 WORDS)
Allyl isothiocyanate, a constituent of mustard and certain vegetables found in the human diet, was tested for cytotoxic and
cytostatic effects in HT29 human colon carcinoma cells in vitro. For an exposure time of 24 h, allyl isothiocyanate exhibited a Dq of 0.32 μg/ml and a D0 of 0.74 μg/ml. Following detransformation of the cells by treatment with sodium butyrate or dimethylformamide the cells became
more resistant to the cytotoxic effects of allyl isothiocyanate, the Dq increasing to 0.74 μg/ml and the D0 to 0.96 μg/ml (with butyrate) or 0.84 μg/ml (with dimethylformamide). At the Dq value for detransformed cells the survival of the control cells was reduced to 56%. Allyl isothiocyanate was also found to
be less cytostatic to the mass growth of detransformed populations in that daily doses of 1.6 μg/ml over a week reduced the
final number of detransformed cells relative to untreated cultures by <25% whilst growth of the transformed cultures was reduced
by >60%. Given this increased sensitivity of the cells to allyl isothiocyanate when in the transformed state, it is hypothesized
that, when consumed in the human diet, this compound may protect against the development of colorectal cancer by selectively
inhibiting the growth of transformed cell clones within the gastrointestinal mucosa.
Gemcitabine plus cisplatin (GC) and methotrexate, vinblastine, doxorubicin, and cisplatin (MVAC) were compared in patients with locally advanced or metastatic transitional-cell carcinoma (TCC) of the urothelium.
Patients with stage IV TCC and no prior systemic chemotherapy were randomized to GC (gemcitabine 1,000 mg/m2 days 1, 8 and 15; cisplatin 70 mg/m2 day 2) or standard MVAC every 28 days for a maximum of six cycles.
Four hundred five patients were randomized (GC, n = 203; MVAC, n = 202). The groups were well-balanced with respect to prognostic factors. Overall survival was similar on both arms (hazards ratio [HR], 1.04; 95% confidence interval [CI], 0.82 to 1.32; P = .75), as were time to progressive disease (HR, 1.05; 95% CI, 0.85 to 1.30), time to treatment failure (HR, 0.89; 95% CI 0.72 to 1.10), and response rate (GC, 49%; MVAC, 46%). More GC patients completed six cycles of therapy, with fewer dose adjustments. The toxic death rate was 1% on the GC arm and 3% on the MVAC arm. More GC than MVAC patients had grade 3/4 anemia (27% v 18%, respectively), and thrombocytopenia (57% v 21%, respectively). On both arms, the RBC transfusion rate was 13 of 100 cycles and grade 3/4 hemorrhage or hematuria was 2%; the platelet transfusion rate was four patients per 100 cycles and two patients per 100 cycles on GC and MVAC, respectively. More MVAC patients, compared with GC patients, had grade 3/4 neutropenia (82% v 71%, respectively), neutropenic fever (14% v 2%, respectively), neutropenic sepsis (12% v 1%, respectively), and grade 3/4 mucositis (22% v 1%, respectively) and alopecia (55% v 11%, respectively). Quality of life was maintained during treatment on both arms; however, more patients on GC fared better regarding weight, performance status, and fatigue.
GC provides a similar survival advantage to MVAC with a better safety profile and tolerability. This better-risk benefit ratio should change the standard of care for patients with locally advanced and metastatic TCC from MVAC to GC.
To determine the 10-year outcome of patients with muscle-invasive bladder cancer treated by transurethral resection (TUR) alone.
Of 432 newly evaluated patients with muscle-invasive bladder cancer, 151 were treated by standard radical cystectomy or by definitive TUR, if restaging TUR of the primary tumor site showed no (T0) or only non-muscle-invasive (T1) residual tumor. Patients were followed-up every 3 to 6 months thereafter for a minimum of 10 years and up to 20 years. Primary end points of the study were disease-specific survival, survival with a bladder, frequency of recurrent invasive tumors in the bladder, and survival after salvage cystectomy.
The 10-year disease-specific survival was 76% of 99 patients who received TUR as definitive therapy (57% with bladder preserved) compared with 71% of 52 patients who had immediate cystectomy (P: = .3). Of the 99 patients treated with TUR, 82% of 73 who had T0 on restaging TUR survived versus 57% of the 26 patients who had residual T1 tumor on restaging TUR (P: = .003). Thirty-four patients (34%) relapsed in the bladder with a new muscle-invasive tumor, 18 (53%) were successfully treated with salvage therapy via cystectomy, and 16 patients (16%) died of disease.
Radical TUR for muscle-invasive bladder cancer is a successful bladder-sparing therapeutic strategy in selected patients who have no residual tumor on a repeat vigorous resection of the primary tumor site.
The two most commonly used methods to analyze data from real-time, quantitative PCR experiments are absolute quantification and relative quantification. Absolute quantification determines the input copy number, usually by relating the PCR signal to a standard curve. Relative quantification relates the PCR signal of the target transcript in a treatment group to that of another sample such as an untreated control. The 2(-Delta Delta C(T)) method is a convenient way to analyze the relative changes in gene expression from real-time quantitative PCR experiments. The purpose of this report is to present the derivation, assumptions, and applications of the 2(-Delta Delta C(T)) method. In addition, we present the derivation and applications of two variations of the 2(-Delta Delta C(T)) method that may be useful in the analysis of real-time, quantitative PCR data.
Solanum nigrum L. (SNL) has been traditionally used as a herbal plant, whose fruit is believed to have anti-tumor properties, although the mechanism for the activity remains to be elucidated. In this study, we prepared an ethanol extract from ripe fruits of SNL and investigated the mechanism involved in its growth-inhibitory effect on MCF-7 human breast cancer cells. Results from proliferation assay using tritium uptake showed that the proliferative capacity of MCF-7 cells was strongly suppressed in the presence of SNL ethanol extract. This was further confirmed through MTT assay and trypan blue exclusion experiments, which showed a very close correlation between the SNL extract concentration and the surviving cell numbers. The SNL extract-mediated suppression of cell growth was verified to be apoptotic, based on the appearance of DNA laddering, increase in DNA fragmentation, and low fluorescence intensity in nuclei after propidium iodide staining of the cells. Furthermore, the SNL extract was revealed to be a potential scavenger of hydroxyl radicals and DPPH radicals rather than superoxide anions. Collectively, our findings suggest that SNL fruit extract could be used as an anti-oxidant and cancer chemo-preventive material.
There has been much progress in the discovery of small, organic molecules that inhibit protein-protein interactions, particularly in the field of cancer. Tubulin polymerization represents a classic target whose function can be allosterically modulated by small molecules. Several protein-protein complexes that regulate apoptosis, or programmed cell death, appear to be particularly amenable to inhibition by small molecules, and recently described compounds have helped to characterize Bcl-2, MDM2 and XIAP as drug targets. Additionally, small-molecule antagonists have recently been described for several new targets, including Rac1-Tiam1, beta-catenin-T cell factor (Tcf), and Sur-2-ESX. Not only is the list of protein-protein inhibitors growing, but the inhibitors themselves are moving closer to treating disease.