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Activation of Caspase-9/3 and Inhibition of Epithelial Mesenchymal Transition are Critically Involved in Antitumor Effect of Phytol in Hepatocellular Carcinoma Cells: Antitumor Effect of Phytol in Hepatocellular Carcinoma Cells
Abstract
This study was designed to investigate the antitumor mechanism of Phytol in hepatocellular carcinomas including Huh7 and HepG2 cells in association with caspase dependent apoptosis and epithelial mesenchymal transition (EMT) signaling. Phytol significantly suppressed the viability of Huh7 and HepG2 cells. Also, Phytol significantly increased the sub G1 population and terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick end labeling (TUNEL) positive cells in a concentration dependent manner in Huh7 and HepG2 cells. Consistently, Phytol cleaved poly (adenosine diphosphate-ribose) polymerase (PARP), activated caspase-9/3, and Bax attenuated the expression of survival genes such as Bcl-2, Mcl-1, and c-Myc in Huh7 and HepG2 cells. Of note, Phytol also suppressed typical morphology change of EMT such as loss of cell adhesion and formation of fibroblast like mesenchymal cells in HepG2 cells. Furthermore, Phytol also reversed the loss of E-cadherin and overexpression of p-smad2/3, alpha-smooth muscle actin, and Snail induced by EMT promoter transforming growth factor beta1 in HepG2 cells. Overall, our findings suggest that Phytol exerts antitumor activity via apoptosis induction through activation of caspas-9/3 and inhibition of EMT in hepatocellular carcinoma cells as a potent anticancer candidate for liver cancer treatment. Copyright © 2015 John Wiley & Sons, Ltd.
Copyright © 2015 John Wiley & Sons, Ltd.
... Phytol and its metabolites at concentrations in the physiological range (≤10 μM) can alter pathways involved in carcinogenesis, such as increasing apoptosis and decreasing proliferation (Komiya et al., 1999;Kim et al., 2015;Thakor et al., 2017). Proposed mechanisms by which phytol and its metabolites exert their chemo-preventive effects include inducing mitochondrial dysfunction, oxidative damage and intracellular Ca 2+ deregulation, as well as epigenetic changes such as histone deacetylation (Idel et al., 2002;Schönfeld et al., 2004;Schönfeld and Wojtczak, 2007;Leipnitz et al., 2010;Kruska and Reiser, 2011;Grings et al., 2012;Borges et al., 2015;Dhaunsi et al., 2016;Dhaunsi et al., 2017) (Table 1). ...
... PA, phytanic acid; PRA, pristanic acid. of brain tissues showed effects on membrane depolarization, oxidative damage, mitochondrial reactive oxygen species (ROS) generation and histone deacetylation (epigenetic transcription regulation) at concentrations as low as 80 nM of PA (Schönfeld et al., 2004;Reiser et al., 2006;Rönicke et al., 2009;Leipnitz et al., 2010;Borges et al., 2015;Nagai, 2015). In HepG2 cells, 40 µM of phytol can suppress the epithelial-mesenchymal transition signaling, which is important for tumor invasion (Kim et al., 2015). Conversely, phytol and its metabolites may also have tumor-promoting properties, as ROS generation can induce DNA damage that either causes apoptosis or transforms normal cells into cancerous cells (Gu et al., 2018;Samimi et al., 2018). ...
... In the physiological range (≤10 μM), phytol and its metabolites can be cytotoxic to various cancer cell lines (Idel et al., 2002;Mobley et al., 2003;Kahlert et al., 2005;Reiser et al., 2006;Tang et al., 2007;Rönicke et al., 2009;Che et al., 2013;Pejin et al., 2014;Kim et al., 2015;Nagai, 2015;Thakor et al., 2017), as shown in Table 2. Although the induction period and measures of cytotoxicity slightly vary among studies, the lowest concentrations at which cytotoxicity was observed were 5 µM of PA in neuroblastoma Neuro2a cells (Nagai, 2015) and 8.8 µM of phytol in breast adenocarcinoma MCF-7 (Pejin et al., 2014). ...
This review summarizes the current evidence on the potential role of phytol, a microbial metabolite of chlorophyl A, and its metabolites, phytanic and pristanic acids, in carcinogenesis. Primary food sources in Western diets are the nut skin for phytol and lipids in dairy, beef and fish for its metabolites. Phytol and its metabolites gained interest as dietary compounds for cancer prevention because, as natural ligands of peroxisome proliferator-activated receptor-α and -γ and retinoid X receptor, phytol and its metabolites have provided some evidence in cell culture studies and limited evidence in animal models of anti-carcinogenic, anti-inflammatory and anti-metabolic-syndrome properties at physiological concentrations. However, there may be a narrow range of efficacy, because phytol and its metabolites at supra-physiological concentrations can cause in vitro cytotoxicity in non-cancer cells and can cause morbidity and mortality in animal models. In human studies, evidence for a role of phytol and its metabolites in cancer prevention is currently limited and inconclusive. In short, phytol and its metabolites are potential dietary compounds for cancer prevention, assuming the challenges in preventing cytotoxicity in non-cancer cells and animal models and understanding phytol metabolism can be mitigated.
... Phytol is a common terpenoid of highly aromatic plants such as green tea, mint, tarragon, basil, and cannabis cultivars [109,110]. This terpenoid has been speculated to hold antioxidant [111,112], anti-inflammatory [113], analgesic [112], anti-cancer [111,114] anti-anxiety [115], anti-convulsant [116], and sedative [117] properties. Phytol and its derivatives have also been explored for toxicity in immune-compromised mice, suggesting non-toxic effects [118][119][120]. ...
... Phytol is a common terpenoid of highly aromatic plants such as green tea, mint, tarragon, basil, and cannabis cultivars [109,110]. This terpenoid has been speculated to hold antioxidant [111,112], anti-inflammatory [113], analgesic [112], anti-cancer [111,114] anti-anxiety [115], anti-convulsant [116], and sedative [117] properties. Phytol and its derivatives have also been explored for toxicity in immune-compromised mice, suggesting non-toxic effects [118][119][120]. ...
Cannabis is a complex biosynthetic plant, with a long history of medicinal use. While cannabinoids have received the majority of the attention for their psychoactive and pharmacological activities, cannabis produces a diverse array of phytochemicals, such as terpenes. These compounds are known to play a role in the aroma and flavor of cannabis but are potent biologically active molecules that exert effects on infectious as well as chronic diseases. Furthermore, terpenes have the potential to play important roles, such as synergistic and/or entourage compounds that modulate the activity of the cannabinoids. This review highlights the diversity and bioactivities of terpenes in cannabis, especially minor or secondary terpenes that are less concentrated in cannabis on a by-mass basis. We also explore the question of the entourage effect in cannabis, which studies to date have supported or refuted the concept of synergy in cannabis, and where synergy experimentation is headed, to better understand the interplay between phytochemicals within Cannabis sativa L.
... Phytol is released during ruminal digestion of the green plants in ruminants (Van den Brink and Wanders, 2006) [1] and thus is present at significant levels in meat and dairy products. Phytol is present in vitamin K, vitamin E, and other tocopherols (Kim et al., 2015) [2] . Phytol is used as an aromatic ingredient in many fragrant compounds and also found in some cosmetic and noncosmetic products (McGinty et al., 2010) [3] . ...
... Phytol is released during ruminal digestion of the green plants in ruminants (Van den Brink and Wanders, 2006) [1] and thus is present at significant levels in meat and dairy products. Phytol is present in vitamin K, vitamin E, and other tocopherols (Kim et al., 2015) [2] . Phytol is used as an aromatic ingredient in many fragrant compounds and also found in some cosmetic and noncosmetic products (McGinty et al., 2010) [3] . ...
Phytol is a component of chlorophyll and is abundantly present in nature. Chemically, it is a diterpene and a branched-chain unsaturated acyclic fatty alcohol. Phytol is converted to phytanic acid and pristanic acid through α-oxidation and β-oxidation respectively. Its significant diverse bioactivities have recently drawn attention for their possible application in the pharmaceutical and biotechnological fields. Phytol had large variety of pharmacological actions and its metabolites had an influence in eliciting these actions. Literature search has revealed that phytol and its metabolites had an antidiabetic action and it can be used for the treatment of metabolism related disorders. The present study was undertaken to assess the effect of orally and intravenously administered phytol in healthy rats. Single dose of phytol was given orally and intravenously to separate groups of rats at a dose rate of 250 mg/kg and 25 mg/kg respectively. From the study, it was noted that all the animals were apparently healthy and showed normal ratty behaviour. Phytol as a single oral dose caused a significant increase in the weekly body weight with no significant effect on the feed and water intake. On the contrary, there was an increase in the feed intake of rats with no significant change in the weekly body weight following administration of phytol intravenously. However, administration of phytol through either route did not affect the water intake of rats. The relative organ weights of liver, kidney and small intestine were significantly increased after oral administration. Whereas, the relative organ weight of liver, lung and reproductive organ of rats showed a significant increase following intravenous administration of phytol when compared to that of control rats. Gross pathological lesions were not observed in any of the organs examined. Phytol is safe for therapeutic use by both oral and intravenous routes at the dose administered.
... Phytol is an acyclic diterpene alcohol that derive from metabolism of chlorophyll in some plants (27). Although, some biological activities such as anticancer (28)(29)(30)(31), anti-microbial (32) and anti-inflammatory (33) effects were reported for phytol, its potential to eliminate CSCs and downregulate CSCs markers has not been explored until now. ...
... This process contribute to the acquisition of invasive properties that are necessary for metastasis (52,53) and its molecular hallmark is downregulation of epithelial markers such as E-cadherin and upregulation of mesenchymal markers, including N-cadherin, Vimentin and Snail (52,53). It was shown that phytol reverse loss of E-cadherin and overexpression of p-smad2/3, alpha-smooth muscle actin, and snail in hepatocellular carcinoma cells (29). Given that cells that have undergone an EMT share many features with CSCs, have drug resistant phenotype and express stem cell markers (44,54), it is likely that phytol with anti-EMT properties has inhibitory effect on CSCs and expression of CSC markers. ...
Cancer stem cells (CSCs), a subgroup of cancer cells, have self-renewal capacity and differentiation potential and drive tumor growth. CSCs are highly-resistant to conventional chemo-radio therapy. Phytochemicals were shown to be able to eliminate CSCs. Phytol is a diterpene alcohol with demonstrated anticancer effects. The current study compared the effect of phytol with retinoic acid (RA) as a well-known inducers of CSC differentiation and cisplatin, a common chemotherapy drug, on CSC markers in human embryonic carcinoma NCCIT cells. NCCIT cells were exposed to 10 mM RA for 14 day to induce differentiation. Moreover, NCCIT cells were treated with IC50 dose of cisplatin (12 µM) and phytol (40 µM) for 7 day. Real-time PCR showed that phytol was more effective that RA and cisplatin in down-regulating the CSC markers OCT4, NANOG, SOX2, ALDH1, ABCB1, CD44 and CD133. Percentage of SP (13%) and ABCB1⁺ (0.34%) in NCCIT cells decreased to 7% and 0.1% respectively after treatment with phytol. A very small proportion of NCCIT cells were positive for CD44 (0.2%) and CD133 (0.48%) and this fraction did not change significantly after treatment with three agents. In conclusion, phytol has the greatest inhibitory effect on CSC population and markers than RA and cisplatin.
... Additionally, Chikati [56] suggested that, phytol dose-dependently inhibited the growth of MDA MB 231 cells along with an anti-invasive effect. Furthermore, in Huh7 and HepG2 cells, phytol at 20-100 µM exhibited an antitumor effect via induction of apoptosis through the activation of caspase-9/3 and inhibition of EMT [57]. Phytol induced apoptosis in AGS gastric adenocarcinoma cells, as observed by the increase of cells in the sub G1 mitotic phase, low regulation of Bcl-2, Bax upregulation, activation of caspases 3 and 9, PARP cleavage, depolarization of mitochondrial membranes and induction of autophagy. ...
... Antiproliferative activity: autophagy with an increase in acidic vacuole organelle formation and GFP-LC3 puncta, inhibition of MECL-induced cell death by pre-treatment with the autophagy inhibitor 3-MA [34] Molt 4B cells -Induction of apoptosis [54,55] Cells brain Astrocyte cell 50 mM Phytanic acid-induced death by activating the mitochondrial route of apoptosis [67] Mouse neuroblastoma Neuro2a cells 10 µM Phytanic acid-induced mitochondrial abnormality and cell death via activation of Hdac2,3 [60] Rat cells brain 10-50 µM Phytanic acid-induced oxidative stress [62] Rat fibroblasts -Phytanic acid-induced lipid accumulation and cytotoxicity [63] Antitumor activity via apoptosis induction through activation of caspase-9/3 and inhibition of EMT in hepatocellular carcinoma cells [57] i.p., intra-peritoneal; p.o., per oral; MAPK, mitogen activated protein kinase; MCP-1, monocyte chemoattractant protein-1; MDA, malonylaldehyde; MIP-1, macrophage inflammatory proteins; MNU, methylnitrosourea; NAD, nicotinamide adenine-di-nucleotide; NLRP, NACHT leucine-rich repeat; NLRP3, PYD domains-containing protein 3; PPARα, peroxisome proliferator-activated receptor alpha; SOD, superoxide dismutase; TCA, tri-carboxylic acid; TIMP, metalloproteinases. metabolism and glucose homeostasis and PPAR-β/δ, which to be involved in anti-inflammatory effect [65,66]. ...
Background:
Phytol have various pharmacological activities such as antimicrobial, cytotoxic, antitumoral, antimutagenic, anti-atherogenic, antidiabetic, lipid-lowering, antispasmodic, antiepileptic, antinociceptive, antioxidant, anti-inflammatory, anxiolytic, antidepressant and immunoadjuvant. Several studies point to an association of phytol with implications for apoptosis and necrosis at cellular levels in cancer, yet no clear conclusions were drawn.
Method:
To clarify this, we conducted a meta-analysis of non-clinical studies of phytol and its associations with toxicity and cytotoxicity emphasizing the mechanisms of apoptosis and necrosis induction and its importance in tumor therapy. Relevant studies were systematically searched in PubMed and Web of Science. The association between phytol and cyto-/toxicity was assessed by odds ratio (ORs) and 95% confidence intervals (CI). Twentythree studies were finally included in the meta-analysis. A significant association between phytol and toxicity (OR: 1.47; 95% CI = 0.86-2.48) was found among in vivo studies and cytotoxicity (OR: 1.81; 95% CI = 1.12- 2.65, p<0.05) in in vitro and ex vivo studies. In in vitro studies, 24% of them indicate that phytol at high doses induces apoptosis by several mechanisms; while about 40% of ex vivo studies indicate that phytol induces reactive oxygen species generation. But, Phytol does not act as a direct oxidant, unlike its metabolite phytanic acid. The 24% of in vivo studies also highlighted the mechanisms for apoptosis-like including expression of Bcl2 protein or mutations in pro-apoptotic protein Bax. Of them, 8% studies show necrosis and hepatotoxicity. However, in 24% of the articles, the mechanisms of toxicity and cytotoxicity are still not well elucidated.
Conclusion:
This study confirms that the association between phytol and cyto-/toxicity depends on the dose/concentration used in the given experimental conditions. Thus, there are still great prospects for new research aimed at the use of phytol and its metabolite as anticancer agents.
... Phytol is a precursor for the synthesis of vitamin K1 and E [19]. Phytol is an acyclic monounsaturated diterpene alcohol used as an aromatic ingredient in many fragrant compounds and also found in some cosmetic and non-cosmetic products [20]. ...
... caspase 3) observed in the present study. The result of significant change in caspase 9 and 3 correlates with the findings of who reported that the activation of Caspase-9/3 is critically involved in antitumor effect of phytol in hepatocellular carcinoma cells [19]. From, gene expression study, it can be said that phytol causes cell death by activation of intrinsic as well as extrinsic apoptosis pathway in human lung carcinoma cells. ...
A number of drugs as well as lead molecules are isolated from natural sources. Phytol is one of such lead molecule belongs to terpenes group distributed widely in medicinal plants. In the present work, we investigated the cytotoxic behavior of phytol on human lung carcinoma cells (A549). Phytol was found to cause characteristic apoptotic morphological changes and generation of ROS in A549 cells. The mechanism of phytol involved the activation of TRAIL, FAS and TNF-α receptors along with caspase 9 and 3. In silico molecular docking studies revealed that phytol has a good binding affinity with glucose-6-phosphate dehydrogenase (G6PD), which is known to promote tumor proliferation. The ability of phytol to become potential drug candidate has been revealed from the pharmacokinetic study performed in the present study.
... Additionally, Chikati [56] suggested that, phytol dose-dependently inhibited the growth of MDA MB 231 cells along with an anti-invasive effect. Furthermore, in Huh7 and HepG2 cells, phytol at 20-100 µM exhibited an antitumor effect via induction of apoptosis through the activation of caspase-9/3 and inhibition of EMT [57]. Phytol induced apoptosis in AGS gastric adenocarcinoma cells, as observed by the increase of cells in the sub G1 mitotic phase, low regulation of Bcl-2, Bax upregulation, activation of caspases 3 and 9, PARP cleavage, depolarization of mitochondrial membranes and induction of autophagy. ...
... Antiproliferative activity: autophagy with an increase in acidic vacuole organelle formation and GFP-LC3 puncta, inhibition of MECL-induced cell death by pre-treatment with the autophagy inhibitor 3-MA [34] Molt 4B cells -Induction of apoptosis [54,55] Cells brain Astrocyte cell 50 mM Phytanic acid-induced death by activating the mitochondrial route of apoptosis [67] Mouse neuroblastoma Neuro2a cells 10 µM Phytanic acid-induced mitochondrial abnormality and cell death via activation of Hdac2,3 [60] Rat cells brain 10-50 µM Phytanic acid-induced oxidative stress [62] Rat fibroblasts -Phytanic acid-induced lipid accumulation and cytotoxicity [63] Antitumor activity via apoptosis induction through activation of caspase-9/3 and inhibition of EMT in hepatocellular carcinoma cells [57] i.p., intra-peritoneal; p.o., per oral; MAPK, mitogen activated protein kinase; MCP-1, monocyte chemoattractant protein-1; MDA, malonylaldehyde; MIP-1, macrophage inflammatory proteins; MNU, methylnitrosourea; NAD, nicotinamide adenine-di-nucleotide; NLRP, NACHT leucine-rich repeat; NLRP3, PYD domains-containing protein 3; PPARα, peroxisome proliferator-activated receptor alpha; SOD, superoxide dismutase; TCA, tri-carboxylic acid; TIMP, metalloproteinases. metabolism and glucose homeostasis and PPAR-β/δ, which to be involved in anti-inflammatory effect [65,66]. ...
... Our data suggest that the most relevant compounds in the n-hexane and DCM subfraction of K. pinnata leaf ethanolic extract are phytol, stigmasterol, ethyl palmitate, and ethyl linoleate. Phytol and its metabolites at concentrations in the physiological range (≤ 10 µM) can alter pathways involved in carcinogenesis, such as increasing apoptosis, decreasing proliferation, and inhibiting the cancer stem cell population [64][65][66]. Stigmasterol is a relevant phytosterol in various herbal plants and possesses anticancer activity. Studies demonstrate that stigmasterol inhibits endothelial cell proliferation, migration, and capillary network formation [64]. ...
Background
The leaves of Kalanchoe pinnata (Lam.) Pers. (K. pinnata), a succulent plant native to tropical regions, are used as a medicinal alternative against cancer in several countries worldwide; however, its therapeutic potential to fight cancer has been little addressed. In this study, we analyzed the phytochemical content, antioxidant capacity, and selectivity of K. pinnata leaf ethanolic extract against different human cancer cell lines in vitro.
Methodology
This study subjected the ethanolic extract to enzymatic assays to quantify the phytochemical content (phenolics, flavonoids, and anthraquinones) and its radical scavenging and iron-reducing capacities. Also, the phytoconstituents and major phenolic compounds present in the extract’s subfractions were identified by GC-MS, HPLC, and NMR. Human cancer (MCF-7, PC-3, HT-29) and normal colon (CoN) cell lines were treated with different concentrations of K. pinnata leaf ethanolic extract, and the changes in cell proliferation (sulforhodamine B assay), caspases activity (FITC-VAD-FMK reporter), mitochondrial membrane potential (MMP, rhodamine 123 assay), chromatin condensation/fragmentation (Hoechst 33342 stain), and ROS generation (DCFH2 probe assay) were assessed.
Results
The results showed that the K. pinnata leaf ethanolic extract is rich in phytoconstituents with therapeutic potential, including phenols (quercetin and kaempferol), flavonoids, fatty acid esters (34.6% of the total composition), 1- triacontanol and sterols (ergosterol and stigmasterol, 15.4% of the total composition); however, it presents a poor content of antioxidant molecules (IC50 = 27.6 mg/mL for H2O2 scavenging activity vs. 2.86 mg/mL in the case of Trolox). Notably, the extract inhibited cell proliferation and reduced MMP in all human cell lines tested but showed selectivity for HT-29 colon cancer cells compared to CoN normal cells (SI = 8.4). Furthermore, ROS generation, caspase activity, and chromatin condensation/fragmentation were augmented significantly in cancer-derived cell lines, indicating a selective cytotoxic effect.
Conclusion
These findings reveal that the K. pinnata leaf ethanolic extract contains several bioactive molecules with therapeutic potential, capable of displaying selective cytotoxicity in different human cancer cell lines.
... Gg suppressed the viability of DU145 (human prostate carcinoma) and DLD1 (colon cancer) cells, generating cell cycle arrest in the G phase. Also, it affected HL-60 (human promyelocytic leukemia cells), K562 (chronic myelogenous leukemia), Molt-3 (acute lymphoblastic leukemia) and COLO320 (adenocarcinoma of the colon) by DNA damage, and these processes eventually led to apoptosis through the activation of caspases (3, 8 and 9) and caused the release of cytochrome C [33][34][35][36]. In the case of MTX, the results obtained from the induction of apoptosis on the U-937 cells are in agreement with some studies on its inhibition effects on the enzyme dihydrofolate reductase, thus preventing the conversion of dihydrofolate to tetrahydrofolate. ...
Ilama leaves are an important source of secondary metabolites with promising anticancer properties. Cancer is a disease that affects a great number of people worldwide. This work aimed to investigate the in vivo, in vitro and in silico anticancer properties of three acyclic terpenoids (geranylgeraniol, phytol and farnesyl acetate) isolated from petroleum ether extract of ilama leaves. Their cytotoxic activity against U-937 cells was assessed using flow cytometry to determine the type of cell death and production of reactive oxygen species (ROS). Also, a morphological analysis of the lymph nodes and a molecular docking study using three proteins related with cancer as targets, namely, Bcl-2, Mcl-1 and VEGFR-2, were performed. The flow cytometry and histomorphological analysis revealed that geranylgeraniol, phytol and farnesyl acetate induced the death of U-937 cells by late apoptosis and necrosis. Geranylgeraniol and phytol induced a significant increase in ROS production. The molecular docking studies showed that geranylgeraniol had more affinity for Bcl-2 and VEGFR-2. In the case of farnesyl acetate, it showed the best affinity for Mcl-1. This study provides information that supports the anticancer potential of geranylgeraniol, phytol and farnesyl acetate as compounds for the treatment of cancer, particularly with the potential to treat non-Hodgkin’s lymphoma.
... The diterpene phytol, widely distributed in nature, has been linked with the expression of PPARc, indicating that is an important molecule in managing abnormalities in lipid metabolism (Goto et al., 2005). The cytotoxic activity against several adenocarcinomas has also been reported for phytol (Kim et al., 2015), as well as its antimicrobial, antioxidant, anti-inflammatory, anxiolytic, and antidepressant activities (Islam et al., 2015). The compound manoyl oxide, is a member of triterpenoids and its anticancer activity has been reported (Deepika et al., 2014). ...
High amounts of raspberries are wasted every year, which are rich source of bioactive compounds. Thus, raspberry waste was used to obtain the essential oil by maceration and the oleoresin by supercritical fluid extraction (SFE). Higher amounts of total phenolics (185 mg GAE g⁻¹) and flavonoids (11.0 mg QE g⁻¹) were obtained in oleoresins than in essential oil (131 mg GAE g⁻¹ and 9.0 mg QE g⁻¹, respectively). DPPH radical scavenging was 1350 μm TE g⁻¹ for oleoresins and 890 μm TE g⁻¹ for essential oil. SFE allowed the extraction of compounds not reported before in red raspberry such as a pinocembrin and farnesol. Oleoresin at 20 μg mL⁻¹ induced a decrease in lipid accumulation during the 3T3‐L1 cell differentiation process, but no after cells were differentiated. Red raspberry waste is an important source of bioactives that can be extracted using green technologies to generate high commercial value by‐products.
... Pristanic acid is a product of phytol metabolism (Bobe et al., 2020). Furthermore, phytol has a cytotoxicity effect against some cancer cell lines (Pejin et al. 2014;Kim et al., 2015;Thakor et al., 2017). It also presence in Calotropis gigantea leaves as larvicidal activity against Spodoptera litura larvae (Babu et al., 2016). ...
Larvicidal potential of Hyptis capitata grown in Indonesia has not been extensively studied. Its leaves are extracted with the maceration method using ethanol as the solvent. Furthermore, the ethanolic leaf extract of the plant was used for larvicidal assays against instar III/IV larvae of Culex quinquefasciatus with different concentrations, namely 1000, 500, 250, 125, and 62.5 µg/mL. Fractionation of the extract was carried out by vacuum liquid chromatography, and obtained four fractions, namely fractions F1, F2, F3, F4. Fractions were also used for the larvicidal assay. The constituents of the extract were then analyzed with the GC-MS method to predict the components involved in its toxicity. Larvicidal data obtained were analyzed using regression analysis to determine the LC50 value. Analysis of variance was carried out with one-way ANOVA using Tuckey HSD-test on the SPSS program 26 at 95% confidence and significance P<0.05. Ethanolic leaf extract has a higher level of toxicity than its fraction. Some compounds that were assumed to play a role in its toxicity include pentadecanoic acid, 2,6,10,14-tetramethyl-, methyl ester; 1-heptadecyne; 9-tetradecen-1-ol, acetate; oxyrane, deodecyl-; 9,12,15-octadecatrienal; and 6,11-dimethyl-2,6,10-dodecatrien-1-ol. These finding indicate that the ethanolic leaf extract of H. capitata has the potential to be developed as a biolarvicidal agent against C. quinquefasciatus.
... Phytols detected in the extracts have shown antiproliferative effects and also induced apoptosis in lung cancer cell lines [12]. In HepG2 hepatocellular carcinoma cells, phytols that induced apoptosis were found to induce it through caspase 9/3 activation and also were found to inhibit EMT (epithelial mesenchymal transition) which plays a key role in tumor invasion [13]. Ethyl palmitate and ethyl linoleate present in the extract were also known to have inhibitory effects on NF-κB [14,15]. ...
... when it was in its highest bloom. Recent studies have shown its antioxidant potential [27], antimicrobial [28], anti-inflammatory [29], antitumour [30], and antidiabetic activities [27]. Its derivative, phytane, was detected in July (6.11%, ...
The present study aimed to isolate volatile organic compounds (VOCs) from fresh (FrHSc) and air-dried (DrHSc) Halopteris scoparia (from the Adriatic Sea) by headspace solid-phase microextraction (HS-SPME) and hydrodistillation (HD) and to analyse them by gas chromatography and mass spectrometry (GC–MS). The impact of the season of growth (May–September) and air-drying on VOC composition was studied for the first time, and the obtained data were elaborated by principal component analysis (PCA). The most abundant headspace compounds were benzaldehyde, pentadecane (a chemical marker of brown macroalgae), and pentadec-1-ene. Benzaldehyde abundance decreased after air-drying while an increment of benzyl alcohol after drying was noticed. The percentage of pentadecane and heptadecane increased after drying, while pentadec-1-ene abundance decreased. Octan-1-ol decreased from May to September. In HD-FrHSc, terpenes were the most abundant in June, July, and August, while, in May and September, unsaturated aliphatic compounds were dominant. In HD-DrHSc terpenes, unsaturated and saturated aliphatic compounds dominated. (E)-Phytol was the most abundant compound in HD-FrHSc through all months except September. Its abundance increased from May to August. Two more diterpene alcohols (isopachydictyol A and cembra-4,7,11,15-tetraen-3-ol) and sesquiterpene alcohol gleenol were also detected in high abundance. Among aliphatic compounds, the dominant was pentadec-1-ene with its peak in September, while pentadecane was present with lower abundance. PCA (based on the dominant compound analyses) showed distinct separation of the fresh and dried samples. No correlation was found between compound abundance and temperature change. The results indicate great seasonal variability of isolated VOCs, as well among fresh and dried samples, which is important for further chemical biodiversity studies.
... These results agreed with those of Pardhasaradhi et al. (2005), which showed that treatment of MCF-7 with A. squamosa seed extracts resulted in nuclear condensation, DNA fragmentation, ROS generation, and apoptosis induction via downregulation of the ratio of Bcl-2/Bax. The remarkable anticancer activity of the extracts may due to the high level of germacrene-D on the leaves which exerted anticancer activity against different cell lines, or to the presence of other bioactive compounds which have also been known to have an anticancer activity such as humulene, phytol and/or a combination of these bioactive compounds (Essien et al., 2016;Kim et al., 2015;Pejin et al., 2014). Collectively, methanolic extracts have the highest total phenolic content and germacrene-D, while acetonic extract have the highest total flavonoid content, these differences on the ratio of the bioactive compounds explain the potential differences between the three extracts (Alnemari et al., 2020). ...
Annona squamosa L. is an important medicinal plant used in traditional medicine for the treatment of various diseases. Different parts of A. squamosa L. have various therapeutic effects; however, the anticancer activity of the leaves has not yet been identified. In vitro, MTT, nuclear staining, and LDH assays were used to evaluate cell survival and proliferation in cells exposed to the extracts. The effect of the extracts on cell migration was investigated using a monolayer wound repair assay, and the apoptotic effects were evaluated using flow cytometry. A breast cancer model was used to study the effect of the extract on the tumor size, and the expression of different proliferative and apoptotic markers was evaluated by immunohistochemical analysis. At a concentration of 100 µg/mL, A. squamosa leaf extracts exerted strong antiproliferative and cytotoxic effects against various cell lines. The extracts reduced wound closure and strongly induced apoptosis. In vivo study, rats were sacrificed 24 h after the last injection, and tumor size, as well as the expression of proliferative and apoptotic markers, were observed to be greatly affected by treatment with the extracts. Therefore, A. squamosa leaf extract may be developed as a potential novel drug to treat breast cancer in the future.
... Another terpene dereplicated, phytol (C 20 H 40 O; accurate mass 296.3079) selectively inhibited the growth of the HepG2 cells with an IC 50 value of 78 ± 3.45 µM [24]. Another study showed that phytol exerted antitumor effect in hepatocellular carcinoma cells by activation of caspases 9/3 [25]. The triterpene cucurbitacin E (formula suggested C 32 H 44 O 8 ; accurate mass 556.3036) exhibited antiproliferative action on Hep3B cancer cells through inhibition of Wnt/β-catenin activation [26]. ...
Cissus incisa leaves have been traditionally used in Mexican traditional medicine to treat certain cancerous illness. This study explored the metabolomic profile of this species using untargeted technique. Likewise, it determined the cytotoxic activity and interpreted all data by computational tools. The metabolomic profile was developed through UHPLC-QTOF-MS/MS for dereplication purposes. MetaboAnalyst database was used in metabolic pathway analysis and the network topological analysis. Hexane, chloroform/methanol, and aqueous extracts were evaluated on HepG2, Hep3B, HeLa, PC3, A549, and MCF7 cancer cell lines and IHH immortalized hepatic cells, using Cell Titer proliferation assay kit. Hexane extract was the most active against Hep3B (IC50 = 27 ± 3 μg/mL), while CHCl3/MeOH extract was the most selective (SI = 2.77) on the same cell line. A Principal Component Analysis (PCA) showed similar profiles between the extracts, while a Venn diagram revealed 80 coincident metabolites between the bioactive extracts. The sesquiterpenoid and triterpenoid biosynthesis pathway was the most significant identified. The Network Pharmacology (NP) approach revealed several targets for presqualene diphosphate, phytol, stearic acid, δ-tocopherol, ursolic acid and γ-linolenic acid, involved in cellular processes such as apoptosis. This work highlights the integration of untargeted metabolomic profile and cytotoxic activity to explore plant extracts, and the NP approach to interpreting the experimental results.
... The authors reported that both extracts ultimately reduced the colonogenic survival of MCF-7 and HCT-116 cells, while both extracts showed lower activity against non-tumor cells (VERO). This remarkable anticancer activity may due to the high level of germacrene-D which exerted anticancer activity against different cell lines [36][37][38], or to the presence of other bioactive compounds which have also been known to have an anticancer activity such as humulene, phytol and/or a combination of these bioactive compounds [59,60]. Collectively, these findings may assist the future development of novel drugs for colon cancer therapy. ...
Background
The research and application of plants in food supplements and drugs have attracted great interest. This study aimed to examine the efficiency of several solvents for the extraction of the main compounds from Annona squamosa leaves and to evaluate the antioxidant, antibacterial, and anticancer activities of these extracts.
Methods
Gas chromatography-mass spectrometry was used to screen the bioactive compounds of A. squamosa methanolic extract. The free radical, hydrogen peroxide, and nitric oxide scavenging activities of the extracts were investigated. Furthermore, MTT, nuclear staining, LDH, and monolayer wound repair assays were performed to evaluate the potential anticancer activity of the extracts in colon cancer cells while the antibacterial activity was tested by using a well diffusion assay.
Results
A. squamosa leaves extracts were found to contain several bioactive compounds, of which the majority were sesquiterpenes (C15H24). These extracts exhibited strong antioxidant activity and antibacterial potency against both gram-positive and gram-negative bacteria. Different A. squamosa leaves extracts displayed remarkable antiproliferative, cytotoxic, antimigration, and apoptotic activities in colon cancer cells.
Conclusions
A. squamosa leaves contain major bioactive compounds that inhibit the growth of several types of bacteria and colon cancer cell lines, which demonstrated their efficacy as an alternative source of antibiotics and for the development of novel drugs for colon cancer therapy.
... The reported ameliorating effect of PYT co-administration could be attributed to previous experimental findings that PYT improves hepatic metabolism through significant suppression of α -Amino-β-carboxymuconate-εsemialdehyde decarboxylase (ACMSD), which plays a key role in regulation of NAD biosynthesis mRNA expression in primary rat hepatocytes and increased blood NAD level (Matsuda et al., 2013). Also, Kim et al., (2015) found PYT induced apoptosis through activation of caspas-9/3 and inhibition of epithelial mesenchymal transition in hepatocellular carcinoma cells lines. ...
Schistosomiasis is one of the most prevalent parasitic diseases in tropical and subtropical areas. Pra-ziquantel is the drug of choice but this drug showed its efficacy against adult only with no effect on juvenile stage and there was drug resistance. Natural drugs are safe. In this study phytol was used to evaluate its efficacy on schistosoma mansoni infected mice. Mice were grouped into 4 groups: Control healthy group included 15 mice, Positive control group included 15 infected animals and received no drugs. Group infected and treated with 40 mg/kg and group treated with 80 mg/kg for two successive days at 6 th week post infection Serum and liver homogenate were taken for all , Biochemical , Hematological and Serum hepatic enzymes. This study revealed that phytol administration improved liver enzymes and decreased Tumor necrosis factor-alfa (TNF-α) in dose dependent manner.
... Phytol is diterpene alcohol it has been reported for many pharmacological activities including cytotoxic, anti-tumorous, anti-mutagenic and anti-teratogenic [32] Phytol is screened against several cell lines it was found that it is a concentration-dependent response in all cell lines, demonstrating to be most effective against MCF-7 with IC50 8.79 ± 0.41 μM; the IC50 values towards other tumors cell line ranged from 15.51 to 69.67 μM. It also exerted weak activity against PC-3 cells, (IC50 77.85 ± 1.93 μM) [33] [34] Squalene is the precursor of cholesterol biosynthesis, it has unique physical property and a wide variety of physiological functions such as anticancer, antihypercholesterolemia and antioxidant potential [35] . Phytosterols were reported for their inhibitory effect on lung, stomach, as well as ovarian and breast cancer. ...
... The activity of EtOAc fraction can be related to an active compound or to the synergic action of several compounds. Phytol as the major constituents in the volatile fraction detected by GC/MS was reported by many authors for its in vitro cytotoxic activity in several tumor cell lines using MTT assays [38] and it also proved apoptosis induction in hepatocellular carcinoma cells [39] . Additionally, the antioxidant activity was estimated through the use of DPPH which is a stable nitrogenous free radical compound has violet color which changes to its reduced form yellow color upon reduction, substances which are able to perform this reaction can be considered as antioxidants [40 , 41] . ...
Astragalus L. species were reported for their important biological activities and they were used in the treatment of several diseases. Astragalus sieberi DC. is a wild plant belonging to family Fabaceae and is growing in the Egyptian deserts. The plant was subjected to extraction by 70% methanol to give the total aqueous methanol extract which was fractionated by organic solvent to give three fractions; petroleum ether (Pet.Ether), ethyl acetate (EtOAc) and methanol (MeOH). These fractions were screened for their in vitro cytotoxic activity using colon (HCT-116) and breast (MCF-7) carcinoma cell lines as well as in vitro antioxidant activity by DPPH assay. The EtOAc fraction showed IC50 32.2 and 69.6 (µg/ml) against HCT-116 and MCF-7, compared to doxorubicin with IC50 37.6 and 26.3 (µg/ml), respectively. Moreover, the pet.ether fraction showed a moderate radical scavenging activity at 100 ug/ml with cell viability 57.6%. Phytochemical investigation of the EtOAc and MeOH fractions of the plant revealed isolation and identification of eight compounds. They were identified as 3-nitro-1-propanol (1), 3-nitro-1-propyl-β-D-glucopyranoside (2), kaempferol (3), rhamnocitrin (4), kaempferol 3-O-(2″-α-arabinopyranosyl)-β-glucopyranoside (5), kaempferol 3-O-(6″-α-rhamnopyranosyl)-β-glucopyranoside (6), isorhamnetin 3-O-(2″-α-arabinopyranosyl)-β-glucopyranoside (7) and isorhamnetin 3-O-(6″-α-rhamnopyranosyl)-β-glucopyranoside (8). GC–MS analysis of the Pet.Ether and EtOAc fractions led to identification of twelve compounds, where phtyol (11.55%), 9,12,15-octadecatrienoic acid methyl ester (10.3%) and 16-octadecenoic acid methyl ester (9.45%) are the major compounds detected in the EtOAc fraction as well as N,N-dimethyl-1-dodecanamine (42.36%) and butylated hydroxytoluene (35.96%) were the major compounds detected in the pet.ether fraction. Additionally, LC–ESI–MS analysis of the MeOH fraction was also performed and revealed the tentative identification of kaempferol and isorhamnetin nuclei with high grade of glycosylation for the first time. Keywords: Astragalus sieberi, GC–MS, LC–ESI–MS, Nitro-compounds, Flavonoids, Cytotoxicity
... The findings would suggest a link between caspase-dependent pathway, BDNF secretion and Akt/PI3K/GSK-3β in HCC cells [34]. A recent report assessed the phytol induced apoptosis in hepatocellular carcinoma cells via activation of caspase-9/3 for mitochondrial dependent pathway and inhibition of survival genes such as Bcl-2, Mcl-1, and c-Myc [35]. According to Fadipe et al. lupeol exhibited LD50 of 289.4 µg/ml against HepG2 [36]. ...
Background
Lavandula genus is an important member of Labiatae (Lamiaceae) family. People use this medicinal plant in treatment of various diseases around the world. The objective of this work was to to investigate cytotoxic effect of different fractions like ethanol, ethyl acetate and n-hexane from methanol extract of Lavandula stoechas aerial parts on HepG2 cell line and investigation of the chemical composition of the ethanolic fraction of the Lavandula stoechas aerial parts which in study showed cytotoxic effect against HepG2 cell line.
Methods
Methanol extract was prepared by cold maceration. Its fractions were obtained in solvents of increasing polarity, i.e., hexane, ethyl acetate and ethanol. In order to evaluate the cytotoxic effect of Lavandula stoechas MTT assay methods was used. For this purpose, HepG2 cell line were treated with the above mentioned L. stoechas aerial parts fractions with different concentration 10, 25, 50, 100, 250, 500 and 1000 µg/ml. The cells without treatment were served as control.
Results
The result of this study showed that ethanolic fraction showed major reduction in % cell survival in compared to other fractions. These results showed that Lavandula stoechas ethanol fraction was highly inhibitory. Further, phyto chemical analysis of ethanolic fraction was carried out using GC-MS technique and the reports showed Lupeol, Phytol, α-Cadinol, Lup-20(29)-en-3-one, hydrocoumarin, Lup-20(29)-en-3-one. Other compounds included fatty acids and their esters. Some of these compounds are being first time reported here from this plant.
Conclusion
MTT assay showed morphological changes, which clearly supports Lavandula stoecha ethanolic fraction posses anticancer potency may be due to presence of phytosterols.
... Moreover, by reducing the expression of glutathione, a Reactive Oxygen Species (ROS) scavenger, citral activates apoptosis of breast cancer cells [43], and in combination with curcumin, induces their apoptosis and cell cycle arrest [45]. Furthermore, phytol inhibits the EMT in hepatocellular carcinoma cells [46]. ...
Pavetta indica L. is used in traditional medicine for the treatment of various diseases including hemorrhoids, headache, urinary conditions, ulcerated nose, and dropsy. However, no study has evaluated the anticancer effect of P. indica L. In this study, we found that a methanol extract of the leaves and branches of P. indica L. (MEPI) caused cell-cycle arrest at the sub-G1 phase and induced apoptosis, as indicated by the activation of caspase-8, -3, -7, and c-PARP. Western blotting revealed that MEPI significantly reduced the levels of markers of the epithelial-mesenchymal transition, such as Vimentin, Snail, Slug, and matrix metallopeptidase 9. Notably, the expression of multidrug resistance-associated protein 1 in triple negative breast cancer (TNBC) was significantly decreased by MEPI. Moreover, the co-treatment with MEPI and doxorubicin resulted in a synergistic reduction in cell viability. MEPI also induced radiation sensitization of TNBC cells. Gas chromatography-mass spectrometry analysis revealed that 5,6-dehydrokawain (DK) is the major constituent of MEPI. Interestingly, DK exerted significant anti-invasive and anti-metastatic effects. Our results provide a strong rationale for investigating the molecular mechanisms of action of MEPI in TNBC.
... Phytol is a component of chlorophyll, which exhibits anticancer and immune-enhancing effects [30]. Phytol induced apoptosis through activation of caspase 9 and 3 in hepatocellular carcinoma cells [31] and lung carcinoma cell line A549 [32]. Phytol augments the activity of natural killer cells to remove cancer cells and increase immunity by regulating macrophage function [30]. ...
Objective: To identify the phytochemical compounds from Annona muricata (A. muricata) and to determine their in vitro anti-proliferative activities against breast cancer cells, MCF7 and MDA-MB-231. Methods: A. muricata leaves were successively extracted by soxhlet method using n-hexane, ethyl acetate and methanol, and decocted with water. Each extract was analysed by gas chromatography mass spectrometry (GCMS) and characterized with Wiley and NIST library searches. Anti-proliferative activity of each extract was evaluated on MCF7 and MDA-MB-231 breast cancer cells using MTT assay. Results: The GCMS analysis of different solvent extracts of A. muricata leaves showed presence of different chemical groups of compounds such as steroids, terpenoids, phenolic compounds, sugars, sugars alcohol and others including vitamin E. Ethyl acetate leaves extract exhibited the lowest IC50 value on the MDA-MB-231 breast cancer cell and n-hexane leaves extract showed the the lowest IC50 value on the MCF-7 breast cancer cell. Conclusion: Steroids and phenolic compounds were the main phytocompound groups identified from all A. muricata leaves extracts. The antiproliferative activity of n-hexane and ethyl acetate extract towards breast cancer MCF7 and MDA-MB-231 respectively might be due to the presence of biologically active compounds in the extracts, hence, providing some scientific evidences of the effectiveness of its traditional usages.
... In EPP, phytol-an acyclic monounsaturated diterpene alcohol and constituent of chlorophyll-was found in a remarkable amount. The anticancer activity of phytol against several tumor cell lines in vitro has been assessed [30][31][32], as well as its capacity to induce the apoptosis in hepatocellular carcinoma cells [33] and in human gastric adenocarcinoma AGS [34]. ...
Recently, seaweeds and their extracts have attracted great interest in the pharmaceutical industry as a source of bioactive compounds. Studies have demonstrated the cytotoxic activity of macroalgae towards different types of cancer cell models, and their consumption has been suggested as a chemo-preventive agent against several cancers such as breast, cervix and colon cancers. Reports relevant to the chemical properties of brown algae Padina sp. are limited and those accompanied to a comprehensive evaluation of the biological activity on osteosarcoma (OS) are non existent. In this report, we explored the chemical composition of French Polynesian Padina pavonica extract (EPP) by spectrophotometric assays (total phenolic, flavonoid and tannin content, and antioxidant activity) and by gas chromatography-mass spectrometry (GC-MS) analysis, and provided EPP lipid and sterols profiles. Several compounds with relevant biological activity were also identified that suggest interesting pharmacological and health-protecting effects for EPP. Moreover, we demonstrated that EPP presents good anti-proliferative and pro-apoptotic activities against two OS cell lines, SaOS-2 and MNNG, with different cancer-related phenotypes. Finally, our data suggest that EPP might target different properties associated with cancer development and aggressiveness.
... Therefore, in response to DNA damage, cells can activate PARP1 as a mechanism of accessibility of DNA repair enzymes and transcriptional factors (58), with relevance for damage repair and cell survival (59), though studies indicate that PARP1 is implicated in apoptosis (60) or necrosis (61). A number of published papers indicated that PHY and its metabolite directly induce expression of PPAR-dependent luciferase activity in HepG2 cells, independent of its phytanic acid metabolite (62), cleave the PARP, and activate caspase 9/3 and Bax that attenuate the expression of Bci2, Mcl-1, and c-Myc genes (63). ...
Phytol (PHY) (3,7,11,15-tetramethylhexadec-2-en-1-ol) exhibits various pharmacological properties including toxicity and cytotoxicity, and exerts antitumor activity. Owing to the urgent need of new pharmaceutical formulations for breast cancer therapy, this study aimed at the evaluation of antitumor activity of PHY in 7,12-dimethylbenzanthracenecancer- induced animal model. Comet assay was employed to evaluate the cytogenetics, DNA repair, and antigenotoxic activities of PHY in neoplastic (breast) and non-neoplastic rodent cells (bone marrow, lymphocytes, and liver). Additionally, hematological, biochemical, histopathological, and immunohistochemical analyses were carried out in experimental animals. Thirty nonpregnant female mice (n = 5) underwent 7 weeks treatment with 6 mg/kg pro-carcinogen, PHY (4 mg/kg), and cyclophosphamide (25 mg/kg). Induction of cancer was confirmed by histopathology and immunohistochemistry for Ki-67. Results suggest that PHY exhibits low toxicity in comparison with other groups in hematological, biochemical, histopathological, and organ size parameters. Additionally, PHY showed modulatory effects on the procarcinogen, and induced genotoxicity and apoptosis in breast cancer cells. Furthermore, it showed a DNA damage repair capacity in mouse lymphocytes. These data indicate that PHY may have the potential as an anticancer candidate in pharmaceutical consumption.
... In this context, PA concentration between 1 and 500 μM was shown to cause oxidative damage of cerebellum and cerebral cortex of Wistar male rats (Leipnitz et al., 2010). Some of the bioactivities of PYT which could be related to its redox properties are its reported in vitro anticancer effects (Chikati, 2013;Guo et al., 2014;Hibasami et al., 2002;Kim et al., 2015;Komiya et al., 1999), anti-teratogenic activity (Arnhold et al., 2002), tumor-promotor (Kagoura et al., 1999) and anti-tumor activity (Líška et al., 2011). Depicted in Fig. 3 are proposed PYT-mediated antioxidant mechanisms (Islam et al., 2016b). ...
... 26 A recent study assessed the antitumor activity of phytol in hepatocellular carcinoma (HepG2 and Huh7) cells in association with epithelial mesenchymal transition (EMT) of hepatocytes signaling and caspase-dependent apoptosis. 27 In the MTT test, HepG2 cells were more susceptible to the diterpene than Huh7 cells. Phytol induced apoptosis in hepatocellular carcinoma cell lines with a loss of cell adhesion and formation of fibroblast. ...
Glycyrrhiza glabra cultivation and harvesting produces substantial quantities of aerial parts as waste. With the aim to prospect an innovative valorization of these byproducts, the aerial parts were harvested in May and October and analyzed for their chemical profile, antioxidant properties, and effects on viability of five cancer cell lines. Pinocembrin was the main constituent. A significant protection of lipid peroxidation was observed with the May total extract (IC50 of 4.2 ± 0.4 μg/mL at 30 min of incubation). The effects on viability of HeLa, MCF-7, MDA-MB-231, Caco-2, and PC3 human cancer cells were investigated. All samples shown a remarkable activity with IC50 values below 25 μg/mL. Samples from plants harvested in May exhibited greater activity than those harvested in October. MCF-7 and HeLa were the most sensitive cells with IC50 in the range 2.73–3.01 and 3.28–5.53 μg/mL, respectively. G. glabra aerial parts represent a good source of valuable products.
... The preliminary phytochemical study identified 31 known compounds using GC-MS analysis. Out of the 31 identified phytochemicals, vanillin (Lirdprapamongkol et al., 2005;Ho et al., 2009;Lirdprapamongkol et al., 2009), phytol (Kim et al., 2015), squalene (Murakoshi et al., 1992;Rao et al., 1998;Smith et al., 1998;Warleta et al., 2010), tetracosane (Uddin et al., 2012), vitamin E (Prasad and Edwards-Prasad, 1982;Jiang et al., 2004;Birringer et al., 2010;Torricelli et al., 2013;Wang et al., 2015), stigmasterol (Kasahara et al., 1994;Kim et al., 2014;Ali et al., 2015) and beta-sitosterol (Awad et al., 1996;Awad et al., 1998;von Holtz et al., 1998;Awad et al., 2000;Awad et al., 2003;Ju et al., 2004;Jourdain et al., 2006;Awad et al., 2007;Koschutnig et al., 2009;Zhao et al., 2009;Baskar et al., 2010) had existing anticancer evidence . However, as this study only detected and not tested the isolated phytoconstituents for their respective anticancer properties, further study is required to identify the exact bioactive compound responsible for the observed anticancer effect and elucidate the involved mechanism. ...
Background
Clinacanthus nutans (C.nutans) is a plant consumed as a cancer treatment in tropical Asia. Despite the availability of numerous anecdotal reports, evaluation of active anticancer effects has remained elusive. Therefore we here examined antiproliferative, reactive oxygen species (ROS)-inducing and apoptosis mechanisms of whole plant extracts in different cancer cell lines.
Methods
Antiproliferative actions of five solvent extracts (hexane, chloroform, ethyl acetate, methanol and water) of C.nutans were tested on non-small cell lung cancer (A549), nasopharygeal cancer (CNE1) and liver cancer (HepG2) cells using MTT assay. The most potent anticancer extract was then assessed by flow cytometry to study cell cycle changes. Intracellular levels of ROS were quantified by DCFH-DA assay. Involvement of the caspase pathway in induction of apoptosis was assessed using caspase assay kits. GC-MS analysis was performed to identify phytoconstituents in the extracts.
Results
Hexane and chloroform extracts were antiproliferative against all three cell lines, while the ethyl acetate extract, at 300 µg/mL, was antiproliferative in the CNE1 but not A549 and HepG2 cases. Methanol and water extracts did not inhibit cancer cell proliferation. The most potent anticancer hexane extract was selected for further testing. It induced apoptosis in all three cell lines as shown by an increase in the percentage of cell in sub-G1 phase. Dose-dependent increase in ROS levels in all three cell lines indicated apoptosis to be possibly modulated by oxidative stress. At high concentrations (>100 µg/mL), hexane extracts upregulated caspases 8, 9 and 3/7 across all three cell lines. GC-MS analysis of the hexane extract revealed abundance of 31 compounds.
Conclusion
Among the five extracts of C.nutans, that with hexane extract demonstrated the highest antiproliferative activity against all three cancer cell lines tested. Action appeared to be via ion of intracellular ROS, and induction of apoptosis via intrinsic and extrinsic caspase pathways.
... The 7 alpha-acetoxyroyleanone, horminone, royleanone, 7-ketoroyleanone and sugiol from Peltodon longipes were known to cause apoptosis and cell cycle arrest at G1/G0 and S phases in human pancreatic cancer cell line MIA PaCa-2 (Fronza et al., 2012), while 7-ethoxyrosmanol arrested G2/M phase and exerted anapoptotic cell death via caspase-3 and caspase-9 dependent pathway in human neuroblastoma cells (Tabata et al., 2012). The chlorophyll-derived diterpenoid, phytol, was found to exert an antitumor activity via apoptosis induction through an activation of caspase-9/3 and inhibition of epithelial mesenchymal transition signalling in HCC cells (Huh7 and HepG2) (Kim et al., 2015a). The apoptosis and cell cycle arresting events are also evident with longikaurin A (Zou et al., 2013), xerophilusin B (Yao et al., 2015a), indolo[3,2-β] andrographolide (Song et al., 2015), tanshinone IIA (Munagala et al., 2015), sclareol (Wang et al., 2015h), ponicidin (Du et al., 2015) and (5R,8R,9S,13R)-halim-1,10-ene-15,16-diol (Silva et al., 2015). ...
As therapeutic tools, diterpenes and their derivatives have gained much attention of the medicinal scientists nowadays. It is due to their pledging and important biological activities. This review congregates the anticancer diterpenes. For this, a search was made with selected keywords in PubMed, Science Direct, Web of Science, Scopus, The American Chemical Society and miscellaneous databases from January 2012 to January 2017 for the published articles. A total 28, 789 published articles were seen. Among them, 240 were included in this study. More than 250 important anticancer diterpenes and their derivatives were seen in the databases, acting in the different pathways. Some of them are already under clinical trials, while others are in the nonclinical and/or pre-clinical trials. In conclusion, diterpenes may be one of the lead molecules in the treatment of cancer. Copyright © 2017 John Wiley & Sons, Ltd.
... In a recent study, phytol alcohol has shown anticancer activity in hepatocellular carcinoma cells (Kim et al., 2015). Phytol's cytotoxicity has also been reported against several other cancer cell lines (Pejin et al., 2014). ...
Different solvent extracts of Dichotomaria obtusata (J. Ellis & Solander) Lamark, Galaxauraceae, a red algae collected from the coast of Bushehr in the Persain Gulf, was investigated for its cytotoxic properties and chemical constituents. The fresh alga, after extraction with methanol and dichloromethane were combined and partitioned between water, dichloromethane and ethyl acetate. The above fractions were then tested against MOLT-4 (human lymphoblastic leukemia) cancer cell line. The IC50 values of the dichloromethane and ethyl acetate layers of the crude extract were 29.8 ± 3.1 and 30.6 ± 7.9 μg/ml against MOLT-4 cells, respectively, while the water layer showed a week activity with IC50 > 50 μg/ml. After fractionation of the active extracts using open column chromatography over silica gel and preparative thin layer chromatography purification, two terpenoid derived compounds, trans-phytol palmitate and γ-tocopherol were isolated from the dichloromethane and ethyl acetate extracts. The structures of the compounds were elucidated using different spectral data including 1H NMR, 13C NMR, HSQC, HMBC and EI-MS. The IC50 values of compounds trans-phytol palmitate, γ-tocopherol and an undetermined mixture of compounds (F-13-14) were determined as 43.4 ± 1.6, – and 20.3 ± 6.2 μg/ml against LS180 (human colon adenocarcinoma); 53.2 ± 9.3, >100 and 27.6 ± 6.9 μg/ml against MCF-7 (human breast adenocarcinoma) and 40.0 ± 4.1, 48.8 ± 1.8 and 15.9 ± 0.3 μg/ml against MOLT-4 cell lines, respectively, which were comparable to the IC50 values of standard anticancer agent, cisplatin against the same cell lines. The red algae collected from the Persian Gulf contained substances that could inhibit the growth of human cancer cell lines and may represent a natural source for the discovery of novel anticancer agents.
... Phytol, a constituent of chlorophyll, is a precursor for the manufacture of synthetic forms of vitamin E [42] and vitamin K1 [51]. Extensive studies have proven anticancer activities of phytol [52][53][54][55]. The catechol substance, 3,5-ditert-butylcatechol, was identified to be an effective inhibitor of the enzyme sarco/endoplasmic reticulum calcium ATPase (SERCA), a potential target for cancer chemotherapy, by virtual screens and confirmed by bioassays [56]. ...
Gynostemma pentaphyllum
(Thunb.) Makino (GpM) has been widely used in traditional Chinese medicine (TCM) for the treatment of various diseases including cancer. Most previous studies have focused primarily on polar fractions of GpM for anticancer activities. In this study, a nonpolar fraction EA1.3A from GpM showed potent growth inhibitory activities against four cancer cell lines with IC
50
ranging from 31.62
μ
g/mL to 38.02
μ
g/mL. Furthermore, EA1.3A also inhibited the growth of breast cancer cell MDA-MB-453 time-dependently, as well as its colony formation ability. EA1.3A induced apoptosis on MDA-MB-453 cells both dose-dependently and time-dependently as analyzed by flow cytometry and verified by western blotting analysis of apoptosis marker cleaved nuclear poly(ADP-ribose) polymerase (cPARP). Additionally, EA1.3A induced cell cycle arrest in G0/G1 phase. Chemical components analysis of EA1.3A by GC-MS revealed that this nonpolar fraction from GpM contains 10 compounds including four alkaloids, three organic esters, two terpenes, and one catechol substance, and all these compounds have not been reported in GpM. In summary, the nonpolar fraction EA1.3A from GpM inhibited cancer cell growth through induction of apoptosis and regulation of cell cycle progression. Our study shed light on new chemical bases for the anticancer activities of GpM and feasibilities to develop new anticancer agents from this widely used medicinal plant.
... The enzyme, aromatase catalizes the biosynthesis of estrogen [56]. According to Guo et al. [57] trans-PYT acting as an inhibitor of aromatase substantiates its anticancer potential (IC 50 : 1 m-mole), although previously PYT has manifested antimutagenic (120 mg/kg; p.o.) specifically by p38 mitogen-activated protein kinases (MAPK) underpinning action [58], anti-teratogenic (500 mg/kg; p.o.) upon inhibitory effects on RAR and RXR receptors to which retinoic acids bind preciously and produce teratogenic effects to wide variety of animals [59], antimammary (500 mg/kg; i.p.) [60], antitumor in hepatocellular carcinoma cells [21] while PA by anti-teratogenic (100 mg/kg; p.o.) [7,59,61] and anti-mammary (with a vitamin D analog) cancer activities [60] in mice. There is also evidence of prostate cancer due to elevated levels of PA [62e65], but the later conclusion is a bit controversial [64,66e68]. ...
In this study essential oil from Oldenlandia diffusa (aerial parts) was extracted using hydrodistillation method. Characterization of the essential oil was done by using GC-MS analysis and identified 71 compounds. Pentacosane (13.29%), hexacosane (11.59%), tetracosane (11.18%), heptacosane (9.76%),tricosane (6.90%), phytol (5.71%), hexatriacontane (4.87%) and isophytol (4.69%) were the major compounds constituting the oil. Further, cytotoxicity of the extracted oil was observed against PA1 (Ovarian), MIAPaCa-2 (Pancreatic), A549 (Lung), MCF7 (Breast), HeLa (Cervical), HepG2 (Liver), PC-3 (Prostatic), MDA-MB-231 (Breast) cell lines. The oil exhibited dose and time dependent inhibition effects against the cancer cell lines. Best inhibition activity was observed against PA1, HeLa and PC-3 cancer cell lines. The IC50 values ranged from 24.19±0.837 - 3.12±0.126 µg/mL in PA1 cells, 51.87±3.104 - 28.95±0.76 µg/mL in HeLa cells and 52.92±1.233 - 14.62±0.465 µg/mL in PC-3 cells at 24, 48 and 72h. From the experiments, it was clear that essential oil of Oldenlandia diffusa should be further explored as an anticancer agent for developing medicinal drug.
Phytol (Pyt), a diterpenoid, possesses many important bioactivities. This study evaluates the anticancer effects of Pyt on sarcoma 180 (S-180) and human leukemia (HL-60) cell lines. For this purpose, cells were treated with Pyt (4.72, 7.08, or 14.16 μM) and a cell viability assay was performed. Additionally, the alkaline comet assay and micronucleus test with cytokinesis were also performed using doxorubicin (6 μM) and hydrogen peroxide (10 mM) as positive controls and stressors, respectively. Results revealed that Pyt significantly reduced the viability and rate of division in S-180 and HL-60 cells with IC50 values of 18.98 ± 3.79 and 1.17 ± 0.34 μM, respectively. Pyt at 14.16 μM exerted aneugenic and/or clastogenic effects in S-180 and HL-60 cells, where the number of micronuclei and other nuclear abnormalities (e.g., nucleoplasmic bridges and nuclear buds) were frequently observed. Moreover, Pyt at all concentrations induced apoptosis and showed necrosis at 14.16 μM, suggesting its anticancer effects on the tested cancer cell lines. Taken together, Pyt showed promising anticancer effects, possibly through inducing apoptosis and necrosis mechanisms, and it exerted aneugenic and/or clastogenic effects on the S-180 and HL-60 cell lines.
Objective:
The aim of this study was to investigate the relationship between miR-767-3p and hepatocellular carcinoma (HCC).
Method:
We examined the expression of miR-767-3p in both HCC tissues and HCC cell lines by qRT-PCR and western blot assay. We also investigated the influence of miR-767-3p on HCC by transfecting HCC cells with either miR-767-3p mimics or inhibitors.
Result:
MiR-767-3p expression was increased in HCCs and cell lines. Functional analyses demonstrated that miR-767-3p increased HCC cell proliferation and prevented apoptosis both in vitro and in vivo, whereas miR-767-3p inhibition had the opposite effect. Caspase-3 and caspase-9 were found to be direct targets of miR-767-3p in HCC cell lines, and overexpression of miR-767-3p suppressed caspase-3/-9 production. Caspase-3 and caspase-9 siRNA knockdown revealed similar promoting of cell proliferation and inhibiting of cell apoptosis produced by miR-767-3p overexpression, whereas caspase-3/-9 siRNAs inhibited miR-767-3p knockdown-induced inhibition of cell proliferation and promotion of cell apoptosis.
Conclusion:
MiR-767-3p promoted proliferation and prevented apoptosis in human hepatocellular carcinoma (HCC) through inhibiting the caspase-3/caspase-9 pathway.
Phytol (PHY), a diterpenoid, is known for its various bio-pharmacological activities. However, its toxicological profile has yet to be evaluated. The aim of this study was to evaluate the cytogenotoxicological profile of PHY in Wistar albino rats. Forty-five female non-pregnant rats were treated acutely and subchronically with PHY at doses of 300 and 2000 mg/kg and 30, 60 and 90 mg/kg for 14 and 28 days. Neuropharmacological, genotoxic, and mutagenic effects were investigated. The results suggest that PHY did not cause the death of rats at a dose of 2000 mg/kg, suggesting a higher range of the LD 50 of this diterpenoid. Several toxicological alterations were observed in clinical and neuropharmacological parameters depending upon the doses. No hepatic histopathological changes were observed. PHY induced genotoxicity in peripheral blood, bone marrow, liver, and kidney. PHY did not show damage repair activity in peripheral blood lymphocytes. In the bone marrow, both acute and subchronic PHY treatments increased micronucleus frequency, indicating a mutagenic effect. PHY caused neuropharmacological alterations and genetic instability, possibly through the oxidative stress induction pathway.
Baccaurea ramiflora Lour. syn. Baccaurea sapida (Roxb.) Muell. Arg. widely known as Burmese grape is native to Southeast Asia. Its leaves, fruits, stem, bark, seeds forms an ingredient in many herbal prescriptions which have been used to treat jaundice, constipation, indigestion, cellulitis, as antidote for sanke venome, antiphlogistic and anodyne against rheumatoid arthritis etc. In the recent years, this plant has been largely
explored on scientific grounds to identify the chemical constituents and pharmacological activities. The present review work is an effort to revisit the scientific works done to evaluate the scope for bio-prospection of B.
ramiflora. Based on the study designed, a number of research papers were reviewed, but only about 35 articles having information on B. ramiflora were evaluated in detail. In total, thirty compounds have been isolated and characterised so far from different parts of this evergreen tree, which accounts for its myriad medicinal value including analgesic, anthelmintic, antioxidant, anti-diarrheal, anti-inflammatory, cytotoxic, haemolytic,
hpoglycemic, hypolipidemic, insecticidal, neuropharmacological, thrmobocytic, anti-fungal and antimicrobial activities. This compilation of assorted information underpins the basic perceptive of B. ramiflora and opens up new horizon for further phytochemical evaluation, safety efficacy, and clinical trials.
The phytocompounds in crude solvent extracts of Pereskia bleo leaves were identified and their cytotoxic effects on cancer cell lines were determined. Crude extracts were obtained via maceration and subjected to GCMS analysis. Then, each extract was incubated with HeLa, MDA-MB-231, SW480, and NIH/3T3 cell lines for 72 h. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide assay was done to determine IC50 values of each extract. Terpenoids, sterols, alkaloids, fatty acids and phenolic compounds were identified from the crude extracts of P. bleo leaves. Other compounds identified were γ-sitosterol, β-tocopherol, and γ-tocopherol. The ethyl acetate extract had potent cytotoxic effect against HeLa and MDA-MB-231 cancer cells as noted by the lowest IC50 values
Emerging evidence suggests that cinobufagin, an active ingredient in Venenum Bufonis, inhibits cell proliferation in several tumor cells. However, the anti-tumor effect of cinobufagin on nasopharyngeal carcinoma and the underlying molecular mechanisms are still unclear. In this study, we found that cinobufagin significantly inhibits the proliferation of nasopharyngeal carcinoma HK-1 cells. Further analyses demonstrated that cinobufagin induces cell cycle arrest at the S phase in HK-1 cells through downregulating the levels of CDK2 and cyclin E. Moreover, cinobufagin significantly downregulates the protein level of Bcl-2 and upregulates the levels of Bax, subsequently increasing the levels of cytoplasmic cytochrome c, Apaf-1, cleaved PARP1, cleaved caspase-3, and cleaved caspase-9, leading to HK-1 apoptosis. Furthermore, we found that cinobufagin significantly increases ROS levels and decreases the mitochondrial membrane potential in HK-1 cells. Collectively, these data imply that cinobufagin induces cell cycle arrest at the S phase and induces apoptosis through increasing ROS levels, thereby inhibiting cell proliferation in HK-1 cells. Therefore, cinobufagin is a promising bioactive agent that may contribute to the development of treatment strategies of nasopharyngeal carcinoma.
The leaves and stems of Lysiphyllum strychnifolium (Craib) A. Schmitz (Fabaceae family) have been traditionally used in Thailand for detoxification and to treat pesticide poisoning in humans. To uncover novel uses of L.strychnifolium, the possible antiviral properties against avian influenza virus A, strain H5N1, were explored in this study. The ethanolic extracts of L.strychnifolium leaves and stems showed good inhibitory activities against H5N1 neuraminidase. Such activities have not been previously reported for this plant. Aqueous extracts did not show any inhibition. Thereafter, the anti-neuraminidase activities of ethanolic extracts were evaluated using fluorometric determination via a MUNANA-based enzyme inhibition assay. Ethanolic extracts of both stems and leaves showed good inhibitory activities against neuraminidase from Influenza A H5N1 with IC50 values of 55.3 and 70.0 μg/mL, respectively. Moreover, anti-bacterial activities of aqueous and ethanolic extracts of L.strychnifolium leaves and stems were tested using the disc diffusion method. All extracts showed broad antibacterial activities against both Gram positive and Gram negative bacterial strains. Phytochemical constituents of all extracts were identified through Gas Chromatography - Mass Spectrometry (GC-MS) and revealed some compounds such as Methyl-p-hydrozybenzoate; Mome inositol; n-Hexadecanoic acid; Tetradecanamide; (Z)-9-Octadecanamide; 1,2,3,-Benzenetriol; Methylparaben; 4-(4-Hydroxyphenyl)-2-butanone; n-Hexadecanoic acid; Ethyl hexadecanoate; Phytol; (Z)-9-Octadecanoic acid, and Octadecanoic acid. This is the first report of neuraminidase inhibitor against Influenza A H5N1 and antibacterial activities derived from L.strychnifolium extracts. The authors suggest that this plant is an alternative source for treatment against influenza viruses and potential antibiotic agent.
Hepatocellular carcinoma (HCC) is a common malignant cancer and is the third leading cause of death worldwide. Effective treatment of this disease is limited by the complicated molecular mechanism underlying HCC pathogenesis. Thus, therapeutic options for HCC management are urgently needed. Targeting the Wnt/β-catenin, Hedgehog, Notch, and Hippo-YAP signaling pathways in cancer stem cell development has been extensively investigated as an alternative treatment. Herbal medicine has emerged as an initiative therapeutic option for HCC management because of its multi-level, multi-target, and coordinated intervention effects. In this article, we summarized the recent progress and clinical benefits of targeting the above mentioned signaling pathways and using natural products such as herbal medicine formulas to treat HCC. Proving the clinical success of herbal medicine is expected to deepen the knowledge on herbal medicine efficiency and hasten the adoption of new therapies. Copyright © 2016 John Wiley & Sons, Ltd.
Hepatocellular carcinoma diagnosis and treatment has witnessed many major changes and challenges in the past two decades. Increasing incidence of HCC has introduced new monitoring systems and increased the efficacy of screening tests, as well as prognosis of the disease, including the staging system, serological testing and diagnostic imaging. Moreover, surgical resection, liver transplantation and herbal therapy have improved treatment. The most encouraging specific serological marker for HCC is alpha fetoprotein (AFP), which, along with ultrasonography, has improved earlier detection of HCC. Most recently, circulating tumor cell measurement has emerged as a promising tool for the prognosis of HCC. Herbal drugs and herbal composite formula drugs are promising towards the prevention of invasion and proliferation of tumor cells. Chemotherapeutic agents, such as sorafenib, bevacizumab and erlotinib, which target growth factor receptors in signaling pathways, are also used as HCC treatments. Furthermore, radiotherapy is employed in the treatment of unresectable tumors. The present report provides an analysis of the above parameters in the management of HCC.
Background:
Platycodin D (PD), a triterpenoid saponin isolated from the Chinese medicinal herb Platycodonis radix, possesses anti-cancer effects in several cancer cell lines. The aim of this study was to evaluate its anti- cancer activities in hepatocellular carcinoma cells.
Materials and methods:
MTT and colony formation assays were performed to evaluate cell proliferation, along with flow cytometry and Western blotting for apoptosis. Cell adhesion was tested by observing cellular morphology under a microscope, while the transwell assay was employed to investigate the cell migration and invasion.
Results:
PD concentration-dependently inhibited cell proliferation in both HepG2 and Hep3B cells, and significantly suppressed colony formation and induced apoptosis in HepG2 cells. The protein levels of cleaved poly ADP-ribose polymerase (PARP) and Bax were up-regulated while that of survivin was down-regulated after treatment with PD. Moreover, PD not only obviously suppressed the adhesion of HepG2 cells to Matrigel, but also remarkably depressed their migration and invasion induced by 12-O-tetradecanoylphorbol 13-acetate (TPA).
Conclusions:
PD presents anti-cancer potential in hepatocellular carcinoma cells via inducing apoptosis, and inhibiting cell adhesion, migration and invasion, indicating promising features as a lead compound for anti-cancer agent development.
Background:
Inoperable and metastatic hepatocellular carcinoma (HCC) is associated with a poor prognosis and low chemotherapeutic efficiency. Sorafenib is an oral multi-kinase inhibitor exerting its effects via the RAF/ MEK/ERK pathway, vascular endothelial growth factor receptor (VEGFR) and platelet derived growth factor receptor beta (PDGFR-β) tyrosine kinases. Randomized studies have shown a significant contribution of sorafenib to life expectancy and quality of life of cancer patients. The aim of the present study is to evaluate the efficacy and side effects of sorafenib therapy in Turkey.
Materials and methods:
Data for 103 patients (82 males, 21 females) receiving sorafenib therapy in 13 centers from February 2008 to December 2012 were evaluated. Median age was 61 years and median ECOG performance status was 1 (range: 0-2). 60 patients (58%) had hepatitis B, 15 patients (15%) had hepatitis C infection and 12 patients (12%) had a history of alcohol consumption. All of the patients had Child scores meeting the utilization permit of the drug in our country (Child A).
Results:
A total of 571 cycles of sorafenib therapy were administered with a median of four per patient. Among the evaluable cases, there was partial response in 15 (15%), stable disease in 52 (50%), and progressive disease in 36 (35%). Median progression-free survival was 18 weeks and median overall survival was 48 weeks. The dose was reduced only in 6 patients and discontinued in 2 patients due to grade 3-4 toxicity, 18 patients (17%) suffering hand-foot syndrome, 7 (7%) diarrhea, and 2 (2%) vomiting.
Conclusions:
This retrospective study demonstrated better efficacy of sorafenib therapy in patients with advanced HCC compared to the literature while progression-free survival and overall survival findings were comparable. The side effect rates indicate that the drug was tolerated well. In conclusion, among the available treatment options, sorafenib is an efficient and tolerable agent in patients with inoperable or metastatic HCC.
Hepatocellular carcinoma (HCC) is one of the most common malignancies, with an increasing incidence. With advances in surgical techniques and instrumentation and the development of molecular-target drugs, a number of potentially curative treatments have become available. Management of HCC patients depends on the stage of their tumor. Liver resection remains the first choice for very early-stage HCC, but it is being challenged by local ablative therapy. For early-stage HCC that meet the Milan criteria, liver transplantation still offers a better outcome; however, local ablative therapy can be a substitute when transplantation is not feasible. Local ablation is also used as a bridging therapy toward liver transplantation. HCC recurrence is the main obstacle to successful treatment, and there is currently no effective means of preventing or treating HCC recurrence. Transarterial therapy is considered suitable for intermediate-stage HCC, while sorafenib is recommended for advanced-stage HCC. This stage-based approach to therapy not only provides acceptable outcomes but also improves the quality of life of HCC patients. Because of the complexity of HCC, therapeutic approaches must be adapted according to the characteristics of each individual patient. This review discusses the current standards and trends in the treatment of HCC.
Abstract Studies suggest that the traditional applications of Kigelia pinnata leaves have beneficial effects against oxidative stress-mediated diseases and cancers. The pulverized dried leaves of K. pinnata were extracted with hexane, ethyl acetate, and methanol sequentially, and the crude extracts were fractionated by silica gel column chromatography with solvent gradient of increasing polarity. 3-hydro-4,8-phytene, trans-phytol, (9Z,12Z)-methyl octadeca-9,12-dienoate, and two oil fractions were obtained. The chemical compositions of chromatographic fractions were determined using gas chromatography-mass spectroscopy. The structure elucidations of the isolated compounds were based on FTIR, MS, and NMR spectral data analyses. These along with the crude extracts were examined for their antioxidant activities using ferric reducing antioxidant power (FRAP), 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging, and 2,2-azinobis(3-ethyl-benzothiazoline-6-sulfonic acid) (ABTS) assays. Total phenolic contents were also determined. The crude extracts and purified compounds were evaluated on the rhabdomyosarcoma human cancer cell for their cytotoxicity using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) cell viability assays. The methanol extract was richer in phenolics and was most potent as antioxidant and cytotoxic agent among all the substances tested. Among the fractions and pure compounds, the two oil fractions showed more cytotoxicity potency, with IC50s of 143.4±0.5 and 147.9±1.3 ng/mL, which is more significant than the reference standard, cyclophosphamide (165.6±1.0 ng/mL). 3-hydro-4,8-phytene showed lower antioxidant and cytotoxicity potential (IC50=1818±5.2 μg/mL and 171.7±0.8 ng/mL, respectively). Trans-phytol did not show a high cytotoxic power (IC50=769.8±4.3 ng/mL). The comparatively high cytotoxicity index of (9Z, 12Z)-methyl octadeca-9,12-dienoate (IC50=153.3±0.1 ng/mL) indicated that it may be one of the principal cytotoxic agent in the ethyl acetate extract. These results suggest that the leaves of K. pinnata possess tumor cytotoxic potential and could be part of a drug combination for future cancer chemotherapy.
The B-cell lymphoma-2 (Bcl-2) family of proteins regulates the intrinsic, or mitochondrial pathway of apoptosis, the final common mechanism of cell death in response to a variety of physiologic and pharmacologic signals, and plays a central role in AML pathogenesis, prognosis and responsiveness to chemotherapy. Traditionally thought to be an important survival factor for multiple myeloma cells, the anti-apoptotic Bcl-2 family protein myeloid cell leukemia-1 (Mcl-1) has recently been shown in preclinical studies to be critical to the development and maintenance of AML, making it an attractive therapeutic target in this disease. Several characteristics, such as its very short half-life, distinguish Mcl-1 from other anti-apoptotic Bcl-2 family members. Additionally, Mcl-1 levels are regulated by a large number of pathways affecting its transcription, translation and degradation. A variety of approaches exploiting these features has been developed to inhibit directly or indirectly the anti-apoptotic function of Mcl-1. Many of these lend themselves well to combination therapies, leading to striking synergism, at least in preclinical models. In this brief review, we highlight some of the more promising strategies targeting Mcl-1 in AML, with a particular emphasis on rational combinations of novel agents.
Neutrophil granulocytes have the shortest lifespan among leukocytes in the circulation and die via apoptosis. At sites of infection or tissue injury, prolongation of neutrophil lifespan is critical for effective host defense. Apoptosis of inflammatory neutrophils and their clearance are critical control points for termination of the inflammatory response. Evasion of neutrophil apoptosis aggravates local injury and leads to persistent tissue damage. The short-lived prosurvival Bcl-2 family protein, Mcl-1 (myeloid cell leukemia-1), is instrumental in controlling apoptosis and consequently neutrophil lifespan in response to rapidly changing environmental cues during inflammation. This paper will focus on multiple levels of control of Mcl-1 expression and function and will discuss targeting Mcl-1 as a potential therapeutic strategy to enhance the resolution of inflammation through accelerating neutrophil apoptosis.
Two different volatile isolates from the aerial parts of Cardaria draba (L.) Desv., obtained either by hydrodistillation (Extract I) or by CH(2) Cl(2) extraction subsequent to hydrolysis by exogenous myrosinase (Extract II), were characterized by GC-FID and GC/MS analyses. The main volatiles obtained by hydrodistillation, i.e., 4-(methylsulfanyl)butyl isothiocyanate (1; 28.0%) and 5-(methylsulfanyl)pentanenitrile (2; 13.8%), originated from the degradation of glucoerucin. In Extract I, also volatiles without sulfur and/or nitrogen were identified. These were mostly hexadecanoic acid (10.8%), phytol (10.2%), dibutyl phthalate (4.5%), and some other compounds in smaller percentages. Extract II contained mostly glucosinolate degradation products. They originated from glucoraphanin, viz., 4-(methylsulfinyl)butyl isothiocyanate (3; 69.2%) and 5-(methylsulfinyl)pentanenitrile (4; 4.5%), glucosinalbin, viz., 2-(4-hydroxyphenyl)acetonitrile (5; 7.2%), and glucoerysolin, viz., 4-(methylsulfonyl)butyl isothiocyanate (6; 5.0%). Moreover, the volatile samples were evaluated for their antimicrobial activity using the disc-diffusion method and determining minimum inhibitory concentrations (MIC). All volatile isolates expressed a wide range of growth inhibition activity against both Gram-positive and Gram-negative bacteria and fungi. The MIC values varied between 4 and 128 μg/ml.
Hepatocellular carcinoma (HCC) poses a major challenge because of the extreme variability of the clinical outcome, which makes it difficult to properly stage the disease and thereby estimate the prognosis. There is growing evidence that this heterogeneous clinical behavior is attributable to several different biological pathways. A novel approach to mapping these differences is by investigating the epigenetics associated with certain clinical aspects.
Herein, the relevance of these molecular differences in combination with the biological and molecular pathways regulating the clinical outcome will be discussed. Use of a mechanistic and pathogenic approach to clarify the natural history of HCC is not just an academic speculation but should help to develop new therapies and to tailor these therapies to each individual patient.
New biological therapies targeting components of the tumoral or peritumoral microenvironment are crucial to the fight against HCC. However, biological redundancies and the presence of several growth factors, hormones, cytokines, etc., potentially involved in HCC tumor progression make it difficult to assess the best target. Sorafenib, a multi-tyrosine kinase inhibitor, blocks the functions of different growth factors present in the tissue microenvironment. The use of Sorafenib in patients with HCC offers a new approach to the therapy of this disease, stimulating research focusing on the development of drugs based on new molecular and pathogenic insights.
The c-myc gene was discovered as the cellular homolog of the retroviral v-myc oncogene 20 years ago (23, 25, 167). The c-myc proto-oncogene was subsequently found to be activated in various animal and human tumors (37, 39, 42). It belongs to the family of myc genes that includes B-myc, L-myc, N-myc, and s-myc; however, only c-myc, L-myc, and N-myc have neoplastic potential (54, 82, 102, 118, 178). Targeted homozygous deletion of the murine c-myc gene results in embryonic lethality, suggesting that it is critical for development (43). Homozygous inactivation of c-myc in rat fibroblasts caused a marked prolongation of cell doubling time, further suggesting a central role for c-myc in regulating cell proliferation (121).
The frequency of genetic alterations of c-myc in human cancers (42) has allowed an estimation that approximately 70,000 U.S. cancer deaths per year are associated with changes in the c-myc gene or its expression. Given that c-myc may contribute to one-seventh of U.S. cancer deaths, recent efforts have been directed toward understanding the function of the c-Myc protein in cancer biology with the hope that therapeutic insights will emerge. Past efforts, which have contributed significantly to our current understanding of c-myc, are discussed in a number of excellent reviews (23, 29, 37, 40, 44, 52, 66, 82, 94, 102, 118, 125, 132, 145, 178, 182, 186).
Cell death and the subsequent post-mortem changes, called necrosis, are integral parts of normal development and maturation cycle. Despite the importance of this process, the mechanisms underlying cell death are still poorly understood. In the recent literature, cell death is said to occur by two alternative, opposite modes: apoptosis, a programmed, managed form of cell death, and necrosis, an unordered and accidental form of cellular dying. The incorrect consequence is the overlapping of: a) the process whereby cells die, cell death; and b) the changes that the cells and tissues undergo after the cells die. Only the latter process can be referred to as necrosis and represents a process in cell life. In this review, we discuss the excellent basic research developed in this field during last decades and problems that remain to be resolved in defining both experimentally and mechanicistically the events that lead to and characterize cell death.
Vitamin K2 (VK2) has a growth inhibitory effect on various types of cancer cells in vitro, and its efficacy has been demonstrated in clinical applications in a number of patients with leukemia and hepatocellular carcinoma. In this study, the effect of cell growth inhibition and apoptosis induction and the concomitant use of an anticancer agent by VK2 (menaquinone: MK4), on gastric cancer cell lines were examined. When 4 kinds of gastric cancer cells (KATO III, MKN7, MKN74 and FU97) were exposed to MK4, the cell growth was inhibited in an MK4 dose-dependent manner. Morphologically, apoptosis induced by MK4 was recognized in FU97, but only a slight number of apoptotic images was recognized in other cell lines. On the contrary, in all the cell lines, the percentage of APO2.7 positive cells increased significantly in the MK4-treated group as compared to the controls. Caspase-3 activity increased significantly in KATO III and FU97 as compared to the controls, while no significant differences were noted in MKN7 or MKN74. Moreover, in all the cell lines, the percentage of G0/G1-phase cells ( approximately 70% in KATO III and FU97, and > or =80% in MKN7 and MKN74) increased in comparison to the controls, suggesting that cell-cycle arrest had occurred. All of the gastric cancer cell lines were given MK4 in different concentrations and two kinds of anticancer agent, with the result that cell growth was inhibited by the anticancer agent in a dose-dependent manner when it was given with MK4 in concentrations of up to 10 microM. In conclusion, our results demonstrate that the effect of MK4 on apoptosis and cell-cycle arrest differs in differentiated (MKN7, MKN74) and undifferentiated (KATO III, FU97) gastric cancer cell lines, and that MK4 alone or with anticancer agents has an antitumor effect on gastric cancer cell lines.
The process of programmed cell death, or apoptosis, is generally characterized by distinct morphological characteristics and energy-dependent biochemical mechanisms. Apoptosis is considered a vital component of various processes including normal cell turnover, proper development and functioning of the immune system, hormone-dependent atrophy, embryonic development and chemical-induced cell death. Inappropriate apoptosis (either too little or too much) is a factor in many human conditions including neurodegenerative diseases, ischemic damage, autoimmune disorders and many types of cancer. The ability to modulate the life or death of a cell is recognized for its immense therapeutic potential. Therefore, research continues to focus on the elucidation and analysis of the cell cycle machinery and signaling pathways that control cell cycle arrest and apoptosis. To that end, the field of apoptosis research has been moving forward at an alarmingly rapid rate. Although many of the key apoptotic proteins have been identified, the molecular mechanisms of action or inaction of these proteins remain to be elucidated. The goal of this review is to provide a general overview of current knowledge on the process of apoptosis including morphology, biochemistry, the role of apoptosis in health and disease, detection methods, as well as a discussion of potential alternative forms of apoptosis.
Vitamin K2 (MK4) has antitumor effects on various types of cancer cell lines in vitro, and its efficacy has also been reported in clinical applications for patients with leukemia, myelodysplastic syndrome, and hepatocellular carcinoma (HCC). However, details of the mechanism of the antitumor effects of MK4 remain unclear. In the present study, we examined the antitumor effects of MK4 on cholangiocellular carcinoma (CCC) cell lines and its mechanism of action using the HL-60 leukemia cell line that exerts MK4-induced cell growth inhibition via apoptosis induction and cell cycle arrest as a control. MK4 exerted dose-dependent antitumor effects on all three types of CCC cell lines. However, apoptosis occurred in a smaller percentage of cells and there was less cell cycle arrest compared with other cancer cell lines studied previously, which suggested slight MK4-induced cell growth inhibition via apoptosis induction and cell cycle arrest. On the contrary, histopathological fidings showed a large number of cells containing vacuoles in their cytoplasm, and electron microscopic findings showed a large number of cytoplasmic autophagosomes and autolysosomes. These findings suggested evidence of autophagy-related cell death. Fluorescence microscopy following acridine orange staining revealed an increase in the number of cytoplasmic acidic vesicular organelles characteristic of autophagy. Moreover, there were few cells forming autophagic vesicles in the control group, while the percentage of cells containing vacuoles in the MK4-treated group increased with the duration of culture. These results suggested that, unlike in leukemia, gastric cancer, HCC, and other cancer cells, the antitumor effects of MK4 on CCC cells are induced via autophagy formation.
Many features of aging result from the incapacity of cells to adapt to stress conditions. When damage accumulates irreversibly, mitotic cells from renewable tissues rely on either of two mechanisms to avoid replication. They can permanently arrest the cell cycle (cellular senescence) or trigger cell death programs. Apoptosis (self-killing) is the best-described form of programmed cell death, but autophagy (self-eating), which is a lysosomal degradation pathway essential for homeostasis, reportedly contributes to cell death as well. Unlike mitotic cells, postmitotic cells like neurons or cardiomyocytes cannot become senescent since they are already terminally differentiated. The fate of these cells entirely depends on their ability to cope with stress. Autophagy then operates as a major homeostatic mechanism to eliminate damaged organelles, long-lived or aberrant proteins and superfluous portions of the cytoplasm. In this mini-review, we briefly summarize the molecular networks that allow damaged cells either to adapt to stress or to engage in programmed-cell-death pathways.
Studies have shown that diterpenes have anti-inflammatory and redox-protective pharmacological activities. The present study aimed to investigate the anti-inflammatory properties of phytol, a diterpene alcohol, in a mouse model of acute inflammation, and phytol effect on leukocyte recruitment, cytokines levels, and oxidative stress. The anti-inflammatory activities of phytol were assessed by measuring paw edema induced by different inflammatory agents (e.g., λ-carrageenan, compound 48/80, histamine, serotonin, bradykinin, and prostaglandin E2 [PGE2 ]), myeloperoxidase (MPO) activity, peritonitis model and cytokine levels. Further, oxidative stress was evaluated by determining glutathione (GSH) levels and malondialdehyde (MDA) concentration. The results showed that phytol (7.5, 25, 50, and 75 mg/kg) significantly reduced carrageenan-induced paw edema, in a dose-dependent manner. In addition, phytol (75 mg/kg) inhibited compound 48/80-, histamine-, serotonin-, bradykinin- and PGE2 -induced paw edema. It also inhibited the recruitment of total leukocytes and neutrophils; decreased MPO activity, tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) levels, and MDA concentration; and increased GSH levels during carrageenan-induced acute inflammation. These results suggest that phytol attenuates the inflammatory response by inhibiting neutrophil migration that is partly caused by reduction in IL-1β and TNF-α levels and oxidative stress.
Purpose:
To retrospectively compare radiofrequency ablation (RFA) combined with the multikinase inhibitor sorafenib (hereafter, sorafenib-RFA) and RFA alone in the treatment of hepatocellular carcinoma (HCC).
Materials and methods:
Institutional review board approval and informed consent were obtained. Between January 2007 and December 2011, 16 patients (mean age, 72.8 years; age range 52-84 years; 10 men, six women) with HCC tumors less than 3 cm in diameter were included in the sorafenib-RFA group, and 136 patients (mean age, 72.1 years; age range, 51-86 years; 92 men, 44 women) with HCC tumors less than 3 cm in diameter were included in the RFA alone (control) group. Mean diameters of the greatest long-axis dimensions of HCC were 22.8 mm ± 4.6 (standard deviation) in the sorafenib-RFA group and 18.1 mm ± 4.4 in the control group. RFA was performed immediately after the 7-day administration of sorafenib. Propensity score matching analysis was used to adjust for potential biases.
Results:
Fifteen of the 16 patients in the sorafenib-RFA group and 30 of the 136 patients in the control group were selected during propensity score matching. No significant differences between the sorafenib-RFA group (n = 15) and the control group (n = 30) were observed with regard to age, sex, etiology, Child-Pugh class, tumor size, puncture number, needle size, location at the liver margin, or location adjacent to a main vessel. The respective mean diameters of the greatest long- and short-axis dimensions of the RFA-induced ablated area were 46.3 mm ± 10.3 and 33.0 mm ± 6.9 in the sorafenib-RFA group and 32.9 mm ± 7.6 and 25.6 mm ± 5.7 in the control group; both of these dimensions were significantly larger in the sorafenib-RFA group (both P < .001).
Conclusion:
Sorafenib-RFA may be superior to standard RFA alone in the treatment of HCC tumors smaller than 3 cm in diameter.
Hepatocellular carcinoma (HCC) represents the most common liver cancer with an increasing incidence and it accounts for the third most common cause of cancer-related death worldwide. Even though the clinical diagnosis and management of HCC improved significantly in the last decades, this malignant disease is still associated with a poor prognosis. It has to be distinguished between patients with HCCs, which developed from liver cirrhosis, and patients without underlying liver cirrhosis as classification systems, prognosis estimation and therapy recommendations differ in-between. In case of HCC in patients with liver cirrhosis in Europe, treatment allocation and prognosis estimation are mainly based on the Barcelona-Clinic Liver Cancer (BCLC) staging system. Based on this staging system different surgical, interventional radiological/sonographical and non-interventional procedures have been established for the multimodal treatment of HCC. The BCLC classification system represents a decision guidance; however because of its limitations in selected patients treatment allocation should be determined on an individualized rather than a guideline-based medicine by a multidisciplinary board in order to offer the best treatment option for each patient. This review summarizes the current management of HCC and illustrates controversial areas of therapeutic strategies.
Hepatocellular carcinoma arises in patients as a consequence of long-standing preexisting liver illnesses, including viral hepatitis, alcohol abuse, or metabolic disease. In such preexisting liver diseases, TGF-β plays an important role in orchestrating a favorable microenvironment for tumor cell growth and promoting epithelial-mesenchymal transition (EMT). TGF-β signaling promotes hepatocellular carcinoma progression by two mechanisms: first, via an intrinsic activity as an autocrine or paracrine growth factor and, second, via an extrinsic activity by inducing microenvironment changes, including cancer-associated fibroblasts, T regulatory cells, and inflammatory mediators. Although there is an increasing understanding on how TGF-β signaling is associated with tumor progression in hepatocellular carcinoma, it is not clear whether TGF-β signaling is limited to a certain subgroup of patients with hepatocellular carcinoma or is a key driver of hepatocellular carcinoma during the entire tumorigenesis of hepatocellular carcinoma. Inhibitors of the TGF-β signaling have been shown to block hepatocellular carcinoma growth and progression by modulating EMT in different experimental models, leading to the clinical investigation of the TGF-β inhibitor LY2157299 monohydrate in hepatocellular carcinoma. Preliminary results from a phase II clinical trial have shown improved clinical outcome and also changes consistent with a reduction of EMT. Cancer Res; 74(7); 1-5. ©2014 AACR.
The MYC oncoprotein is an essential transcription factor that regulates the expression of many genes involved in cell growth, proliferation, and metabolic pathways. Thus, it is important to keep MYC activity in check in normal cells in order to avoid unwanted oncogenic changes. Normal cells have adapted several ways to control MYC levels, and these mechanisms can be disrupted in cancer cells. One of the major ways in which MYC levels are controlled in cells is through targeted degradation by the ubiquitin-proteasome system (UPS). Here, we discuss the role of the UPS in the regulation of MYC protein levels and review some of the many proteins that have been shown to regulate MYC protein stability. In addition, we discuss how this relates to MYC transcriptional activity, human cancers, and therapeutic targeting.
Sorafenib is a potent inhibitor that targets several kinases associated with tumorigenesis and cell survival. Although sorafenib has been approved for the clinical treatment as single agent, combining sorafenib with other agents has been shown to improve its antitumor efficacy in various preclinical tumor models. ABT-263 is second-generation BH3 mimic that binds to the anti-apoptotic family members Bcl-2, Bcl-xL and Bcl-w and subsequently promotes intrinsic apoptosis in cancer cells.
We have previously demonstrated that ABT-263 sensitizes TRAIL-induced apoptosis in human hepatocarcinoma cells. In the present study, we investigated the effects of ABT-263 treatment combined with sorafenib and found that the two agents displayed significant synergistic anti-tumor effects in various human cancer cells.
ABT-263 enhanced sorafenib-induced apoptosis while sparing non-tumorigenic cells. Despite the observation that ABT-263 treatment combination with sorafenib significantly stimulates intracellular ROS production and subsequent mitochondrial depolarization, it is not sufficient to trigger cell apoptosis. ABT-263 combined with sorafenib treatment displayed a significant decrease in Akt activity, which is at least partly involved in combination treatment-induced apoptosis. Interestingly, we found that Bax and p21 (CIP1/WAF1) play a critical role in regulating the synergistic anti-tumor efficacy of ABT-263 and sorafenib. Additionally, combining ABT-263 with sorafenib demonstrated dramatic efficacy in in vivo tumor xenograft models.
These results suggest that the anti-tumor activity of ABT-263 combined with sorafenib may involve the induction of intrinsic cell apoptosis via inhibition of Akt as well as reduced Bax and p21 expression. Our findings offer a novel effective therapeutic strategy for tumor treatment.
The aberrant activation of Wnt/β-catenin signaling plays an important role in the carcinogenesis and progression of hepatocellular carcinoma (HCC). Therefore, the Wnt/β-catenin signaling molecules are attractive candidates for the development of targeted therapies for this disease. The present study showed that destruxin B (DB) inhibits the proliferation and induces the apoptosis of HCC cells by decreasing the protein expression of anti-apoptotic Bcl-2 and Bcl-xL and increasing the expression of the proapoptotic protein Bax. More importantly, DB also attenuates Wnt-signaling in HCC cells by downregulating β-catenin, Tcf4, and β-catenin/Tcf4 transcriptional activity, which results in the decreased expression of β-catenin target genes, such as cyclin D1, c-myc, and survivin. Furthermore, DB affects the migratory and invasive abilities of Sk-Hep1 cells through the suppression of markers of the epithelial-mesenchymal transition (EMT). A synergistic anti-proliferative and migratory effect was achieved using the combination of DB and sorafenib in Sk-Hep1 cells. In conclusion, DB acts as a novel Wnt/β-catenin inhibitor and reduces the aggressiveness and invasive potential of HCC by altering the cells' EMT status and mobility. DB in combination with sorafenib may be considered for future clinical use for the management of metastatic HCC.
Epithelial-mesenchymal transition (EMT) is an important factor in cancer invasiveness and metastatic progression. During EMT, cancer cells acquire stem cell properties. The role of EMT and stemness in colon cancer has not been fully understood. We aimed to demonstrate the clinical significance of EMT and the stem cell phenotype in colorectal cancer. Two hundred and thirty-one surgically resected colon cancer cases were included in the present study. mRNAs of E-cadherin, TWIST1 and SNAI1 were analyzed by quantitative real-time polymerase chain reaction (qRT-PCR) (n=109). Immunohistochemical staining was performed for six markers (ALDH1, TGF-β1, E-cadherin, β-catenin, TWSIT1 and SNAI1) (n=231). We assessed clinicopathological characteristics according to the expression of the stem cell phenotype and EMT markers. Based on the results of qRT-PCR, TWIST1 and SNAI1 significantly influenced node metastasis (P=0.04 and P=0.02, respectively). High TWIST1 and SNAI1 mRNA expression was associated with poor overall survival according to the univariate analysis (P<0.01 and P=0.01, respectively) and the multivariate analysis (P=0.04 and P=0.04, respectively). ALDH1 expression as detected by immunohistochemical staining was associated with high nodal stage, advanced clinical stage, lymphatic invasion and poor survival (P=0.01, P=0.04, P<0.05 and P<0.01, respectively) and with the expression of TGF-β1 and β-catenin. In conclusion, in human colorectal cancer, the EMT markers TWIST1 and SNAI1 are suggested as important markers of poor prognosis. Their expression is associated with the expression of putative stem cell marker ALDH1, and ALDH1 is associated with the expression of TGF-β1.
Activation of fibroblasts and their differentiation into myofibroblasts, excessive collagen production and fibrosis occurs in a number of bladder diseases. Similarly, conversion of epithelial cells into mesenchymal cells (EMT) has been shown to increase fibroblasts like cells. TGF-β1 can induce the EMT and the role of TGF-β1-induced EMT during bladder injury leading to fibrosis and possible organ failure is gaining increasing interest. Here we show that EMT and fibrosis in porcine bladder urothelial (UC) cells are Smad dependent. Fresh normal porcine bladder urothelial cells were grown in culture with or without TGF-β1 and EMT markers were assessed. TGF-β1 treatment induced changes in cellular morphology as depicted by a significant decrease in the expression of E-cadherin and corresponding increase in N-cadherin and α-SMA. We knocked down Smad2 and Smad3 by Smad specific siRNA. Downregulation of E-cadherin expression by TGF-β1 was Smad3-dependent, whereas N-cadherin and α-SMA were dependent on both Smad2 and Smad3. Connective tissue growth factor (CTGF/CCN2), matrix metalloproteinase-2 and -9 (MMP-2, MMP-9) has been shown to play important roles in the pathogenesis of fibrosis. Induction of these genes by TGF-β1 was found to be time dependent. Upregulation of CTGF/CCN2 by TGF-β1 was Smad3 dependent; whereas MMP-2 was Smad2 dependent. Smad2 and Smad3 both participated in MMP-9 expression. TGF-β1 reprogrammed mesenchymal fibroblast like cells robustly expressed collagen I and III and these was inhibited by SB-431542, a TGF-β receptor inhibitor. Our results indicate that EMT of porcine bladder UC cells is TGF-β1 dependent and is mediated through Smad2 and Smad3. TGF-β1 may be an important factor in the development of bladder fibrosis via an EMT mechanism. This identifies a potential amenable therapeutic target.
Hepatocellular carcinoma (HCC) as the major histological subtype of primary liver cancer remains one of the most common malignancies worldwide. Due to quite complicated the molecular pathogenesis of HCC, the option for effective systemic treatment is quite limited. There exists a critical need to explore and evaluate possible alternative strategies for effective control of HCC. With a long history of clinical use, Chinese herbal medicine (CHM) is emerging as a noticeable choice for its multi-level, multi-target and coordinated intervention effects against HCC. With the aids of phytochemistry and molecular biological approaches, in the past decades many CHM-derived compounds have been carefully studied through both preclinical and clinical researches and have shown great potential in novel anti-HCC natural product development. The present review aimed at providing the most recent developments on CHM-derived anti-HCC compounds, especially their underlying pharmacological mechanisms.
A systematic search of CHM-derived anti-HCC compounds was carried out focusing on literatures published both in English (PubMed, Scopus, Web of Science and Medline) and in Chinese academic database (Wanfang and CNKI database).
In this review, we tried to give a timely and comprehensive update about the anti-HCC effects and targets of several representative CHM-derived compounds, namely curcumin, resveratrol, silibinin, berberine, quercetin, tanshinone II-A and celastrol. Their mechanisms of anti-HCC behaviors, potential side effects or toxicity and future research directions were discussed.
Herbal compounds derived from CHM are of much significance in devising new drugs and providing unique ideas for the war against HCC. We propose that these breakthrough findings may have important implications for targeted-HCC therapy and modernization of CHM.
Thrombosis, both venous and arterial, is a major cause of morbidity and mortality worldwide. Consequently, there is an ongoing search for new antithrombotic drugs, particularly novel antiplatelet agents and anticoagulants. A better understanding of the biochemical pathways involved in platelet activation and coagulation and of the links between these systems and the impact of thrombosis on inflammation has led to the identification of new targets for antithrombotic drugs. This paper focuses on these new targets and new antiplatelet drugs and anticoagulants and describes the major advances in the continuing search for more potent antithrombotic drugs that have limited effects on hemostasis.
To study the inhibitory effect of Fuzheng Yiliu Granule (FYG) on hepatocellular cancer (HCC) and investigate the mechanism mediating its bioactivity.
H22 tumor-bearing ICR mice were treated with FYG [3.6 g/(kg·d)] for 5 days. Tumor volume and tumor weight, percentages of CD3(+), CD4(+), CD8(+), and natural killer (NK) cells in peripheral blood, tumor apoptosis and serum levels of interleukin-2 (IL-2), and tumor necrosis factor-α (TNF-α) were evaluated. FYG-containing serum was prepared from SD rats treated for 7 days [high dose 3.6 g/(kg·d); middle dose 1.8 g/(kg·d); low dose 0.9 g/(kg·d)]. Cell cycle, cell viability, and apoptosis were evaluated after HepG2 cell line was cultured in FYG-containing serum for 48 h. The levels of IL-2 and TNF-α in FYG-containing serum were also determined.
FYG produced a potent antitumor effect (P<0.01) and induced marked apoptosis of the tumor tissue (P<0.05). Mice treated with FYG had higher percentages of CD3(+) and CD4(+) (P<0.05), and more NK cells (P<0.01) in the peripheral blood than those in the animals treated with normal saline. Mice receiving FYG had the highest serum levels of IL-2 and TNF-α (P<0.01). High-dose FYG-containing serum significantly decreased HepG2 cell viability, inhibited cell proliferation (P<0.05), and induced apoptosis (P<0.01). In addition, the levels of IL-2 and TNF-α of high-dose-containing serum were higher than the blank serum (P<0.01).
FYG could inhibit HCC growth by regulating immune function and inducing apoptosis of tumor cells in vivo and in vitro.
Prostatic adenocarcinoma is emerging as a major cause of morbidity and mortality in the male population in the western world. Programmed cell death (apoptosis) in the prostate is activated by hormone ablation and is under the control of several regulating genes including the tumour suppressor gene p53 and the proto-oncogene bcl-2. Bcl-2 belongs to a rapidly expanding family of genes which form two functionally antagonistic groups controlling cell death and survival. Apoptosis regulating genes appear to play an important role in the development and progression of prostatic adenocarcinoma and offer a potential target for future therapeutic strategies.
Vasculogenic mimicry (VM) refers to the unique ability of highly aggressive human tumor cells to form matrix-rich networks de novo when cultured on a three-dimensional matrix, thus mimicking embryonic vasculogenesis. Some studies have shown that tumor hypoxia can promote tumor cells to form vessel-like tubes in vitro and express genes associated with VM. Although, the mechanisms involved in hypoxia-induced VM remain elusive, we hypothesized that the epithelial-mesenchymal transition (EMT) regulator Twist may play a major role in hypoxia-induced VM. We investigated this hypothesis in vitro by pretreating hepatocellular carcinoma cells under hypoxic conditions. Following the hypoxia treatment, the cells formed typical pipe-like VM networks. Moreover, the expression of VM markers was increased. Hypoxia-induced VM was accompanied by the increased expression of Twist. Twist siRNA reversed the effects of hypoxia on VM. These results suggest that the overexpression of Twist correlates to hypoxia-induced VM in hepatocellular carcinoma cells.
Epithelial-mesenchymal transition (EMT) is a physiological process that has been recognized to occur during the progression of an increasingly large number of human diseases, including liver fibrosis, cirrhosis, and hepatocellular carcinoma. The activation of transforming growth factor β (TGF-β) signaling is considered a critical event during EMT, and efforts have been made to screen small molecules that interfere with the TGF-β signaling pathway during EMT. Here we report the identification of sorafenib, a clinical agent that inhibits TGF-β signaling. When applied to AML12 cells and primary hepatocytes, sorafenib strikingly suppressed TGF-β1-induced EMT and apoptosis. Additionally, sorafenib inhibited TGF-β1-induced signal transducer and activator of transcription 3 phosphorylation. We further present in vitro evidence that sorafenib ameliorates the proapoptotic and profibrotic effects of TGF-β1 in mouse primary hepatocytes, suggesting that this drug exerts a protective effect on hepatocytes and has therapeutic potential for the treatment of liver fibrosis.
Epithelial-mesenchymal transition (EMT) is a physiological process occurring in the embryo. In adult organism, EMT could be involved in disease development. In the liver, the possibility that EMT of liver epithelial cells participate to liver fibrosis is increasingly discussed. Furthermore, the involvement of hepatocyte EMT to liver cancer biology has also been documented over the past few years. In this review, we will first describe how EMT participates to embryological development. We will then discuss the involvement of hepatocytes and biliary epithelial cells in liver fibrosis. Finally, we will describe how EMT may impact the metastatic process and resistance to therapy in hepatocellular carcinoma.
Unlabelled:
Epithelial-to-mesenchymal transition (EMT) is predicted to play a critical role in metastatic disease in hepatocellular carcinoma. In this study, we used a novel murine model of EMT to elucidate a mechanism of tumor progression and metastasis. A total of 2 x 10(6) liver cells isolated from Pten(loxp/loxp)/Alb-Cre(+) mice, expanded from a single CD133(+)CD45(-) cell clone, passage 0 (P0), were sequentially transplanted to obtain two passages of tumor cells, P1 and P2. Cells were analyzed for gene expression using microarray and real-time polymerase chain reaction. Functional analysis included cell proliferation, migration, and invasion in vitro and orthotopic tumor metastasis assays in vivo. Although P0, P1, and P2 each formed tumors consistent with mixed liver epithelium, within the P2 cells, two distinct cell types were clearly visible: cells with epithelial morphology similar to P0 cells and cells with fibroblastoid morphology. These P2 mesenchymal cells demonstrated increased locomotion on wound healing; increased cell invasion on Matrigel basement membrane; increased EMT-associated gene expression of Snail1, Zeb1, and Zeb2; and down-regulated E-cadherin. P2 mesenchymal cells demonstrated significantly faster tumor growth in vivo compared with P2 epithelial counterparts, with invasion of intestine, pancreas, spleen, and lymph nodes. Furthermore, P2 mesenchymal cells secreted high levels of hepatocyte growth factor (HGF), which we propose acts in a paracrine fashion to drive epithelial cells to undergo EMT. In addition, a second murine liver cancer stem cell line with methionine adenosyltransferase 1a deficiency acquired EMT after sequential transplantations, indicating that EMT was not restricted to Pten-deleted tumors.
Conclusion:
EMT is associated with a high rate of liver tumor proliferation, invasion, and metastasis in vivo, which is driven by HGF secreted from mesenchymal tumor cells in a feed-forward mechanism.
Apoptosis, an essential and basic biological phenomenon, is regulated in a complex manner by a multitude of factors. Myeloid cell leukemia 1 (Mcl-1), an anti-apoptotic member of the B-cell lymphoma 2 (Bcl-2) family of apoptosis-regulating proteins, exemplifies a number of the mechanisms by which a protein's contribution to cell fate may be modified. The N-terminus of Mcl-1 is unique amongst the Bcl-2 family, in that it is rich in experimentally confirmed and putative regulatory residues and motifs. These include sites for ubiquitination, cleavage and phosphorylation, which influence the protein's stability, localisation, dimerization and function. Here we review what is known about the regulation of Mcl-1 expression and function, with particular focus on post-translational modifications and how phosphorylation interconnects the complex molecular control of Mcl-1 with cellular state.
It has become accepted that cell death is an essential mechanism for the maintenance of an animal's health. But how do cells know when they should die, and how do they do it? A recent conference titled "Programmed Cell Death" was held by the American Association For Cancer Research in Bolton Landing, New York state, USA, on October 19-23, 1996, which focused on the signals that push a cell towards its own self-destruction, and also on the biochemical tools used by these cells in making this ultimate sacrifice. The molecular vocabulary of programmed cell death is only recently being deciphered. At the heart of programmed cell death exists at least one intrinsic program for committing cell suicide, although it is still unclear if there is only one [3]. If only one program exists, it probably contains multiple branches, overlapping pathways, and redundant molecular interactions. One item is clear, however: there exist many mechanisms for inducing cells to suicide.
The exposure of human lymphoid leukemia Molt 4B cells to phytol which was isolated from Lolium multiflorum Lam and identified by MS, and 1H- and 13C-NMR, led to both growth inhibition and the induction of programmed cell death (apoptosis). Morphological change showing apoptotic bodies was observed in the cells treated with phytol. The fragmentation by phytol of DNA to oligonucleosomal-sized fragments that are characteristics of apoptosis was observed to be concentration- and time-dependent. These findings suggest that growth inhibition by phytol of Molt 4B cells results from the induction of apoptosis in the cells.
Hand-foot skin reaction is a distinctive cutaneous side-effect of antineoplastic kinase inhibitor-targeted therapy. Severe hand-foot skin reaction requires postponement of treatment or dose reduction. Histopathological studies of skin toxicity associated with kinase inhibitors are currently unavailable.
To report the clinical and histopathological findings of hand-foot skin reaction produced by the multikinase inhibitor sorafenib.
Nine patients with metastatic carcinoma-seven with renal cell carcinoma (RCC), one with melanoma and one with hepatocellular carcinoma (HCC)-received continuous, oral sorafenib 400 mg twice daily. Hand-foot skin reaction was defined and graded according to National Cancer Institute Common Toxicity Criteria 3.0. Biopsies from lesions of erythematous scaly or blistering skin were obtained from five cases (four RCC and one HCC).
Seven of the nine (78%) patients developed hand-foot skin reaction characterized by well-demarcated, tender, erythematous papules and plaques with greyish blisters or hyperkeratotic, callus-like formations on palmoplantar surfaces and distal phalanges. Skin biopsy of hand-foot skin reaction lesions revealed epidermal acanthosis, papillomatosis, parakeratosis, dispersed dyskeratotic cells and keratinocyte vacuolar degeneration. Other skin toxicities included angular cheilitis, seborrhoeic dermatitis and perianal dermatitis.
The clinical manifestations and histopathological features of sorafenib-induced skin reactions are unique. The most relevant histopathological findings of hand-foot skin reaction include keratinocyte vacuolar degeneration, the presence of intracytoplasmic eosinophilic bodies, and intraepidermal blisters in the stratum malpighii. Further studies are warranted to elucidate the mechanisms of this novel multitargeted kinase inhibitor-associated skin reaction.
We investigated the effect of oleanolic acid, a plant-derived triterpenoid, on insulin secretion and content in pancreatic beta-cells and rat islets. Oleanolic acid significantly enhanced insulin secretion at basal and stimulatory glucose concentrations in INS-1 832/13 cells and enhanced acute glucose-stimulated insulin secretion in isolated rat islets. In the cell line the effects of oleanolic acid on insulin secretion were comparable to that of the sulfonylurea tolbutamide at basal glucose levels and with the incretin mimetic Exendin-4 under glucose-stimulated conditions, yet neither Ca(2+) nor cAMP rose in response to oleanolic acid. Chronic treatment with oleanolic acid increased total cellular insulin protein and mRNA levels. These effects may contribute to the anti-diabetic properties of this natural product.
Apoptosis update: to be, or not to be, and how to arrange the latter
- Sheard MA
Senescence, apoptosis or autophagy? When a damaged cell must decide its path-a mini-review
- Vicencio