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Predictive Role of Nuclear Factor-κB Activity in Gastric Cancer: A Promising Adjuvant Approach With Caffeic Acid Phenethyl Ester
Abstract
The biologic significance of nuclear factor-kappaB (NF-kappaB) activation in human gastric cancer is unclear. We clarify the clinical significance of NF-kappaB activation and its relationship to Helicobacter pylori infection, a well-known pathogenesis of gastric cancer. Moreover, we examine the effects and underlying mechanisms induced by caffeic acid phenethyl ester (CAPE), an inhibitor of NF-kappaB, for gastric carcinoma.
NF-kappaB was located immunohistochemically in 90 human gastric cancer specimens and 50 nonmalignant gastric specimens. The correlations between NF-kappaB activation, pathologic staging, and H. pylori infection were analyzed. We also performed electrophoretic mobility gel shift assay, real-time reverse transcription polymerase chain reaction, and enzyme-linked immunosorbent assay to evaluate the responses of AGS (a gastric adenocarcinoma epithelial cell line) human gastric cancer cells subsequent to H. pylori infection or CAPE treatment.
Nuclear expression of NF-kappaB was significantly more frequently observed in gastric cancer tissues than in nonmalignant gastric tissues (31% vs. 4%, P=0.0001). The activity of NF-kappaB and the expressions of MMP-9, IL-1beta, and IL-8 in AGS cells were activated by H. pylori infection. However, the augmented responses could be significantly reversed by CAPE treatment. Moreover, in vitro studies showed that CAPE inhibits tumor growth and capacity for invasion.
NF-kappaB activation is related to carcinogenesis, tumor aggression, and H. pylori infection with the increased expression of MMP-9, IL-1beta, and IL-8. Moreover, NF-kappaB inhibitors or anti-inflammatory agents such as CAPE might be new adjuvant agent against invasive gastric carcinoma.
... NF-κB is a critical mediator of TM expression [21] and transcription of inflammatory cytokines [28], while CAPE significantly attenuated NF-κB activity and suppressed the [29,30]. In the present study, we examined the effects of CAPE on TM expression and tumor aggressiveness of bladder cancer cells. ...
... CAPE, an active anti-inflammatory component of propolis, is a specific inhibitor of NF-κB [42]. CAPE has been reported to significantly inhibit the activation of NF-κB and attenuate proinflammatory cytokine production in cancer cells [29,30,43]. In the present study, we found that attenuated NF-κB activation by CAPE treatment was associated with increased expression of TM in bladder cancer cells. ...
Background
The identification of potential tumor markers will help improve therapeutic planning and patient management. Thrombomodulin (TM) is a sensitive urothelial marker. TM was reported to be one of the endogenous anti-metastatic factors and has diagnostic and prognostic values for the progression of carcinoma. In the present study, we examine the role of TM in bladder cancer.
Methods
We studied the role of TM in tumor behavior and related signaling pathways in vitro using the human bladder cancer cell lines HT1376, HT1197, J82 and T24, and in vivo using animal models. We also selected clinical specimens from 100 patients with bladder cancer for immunohistochemical staining to evaluate the predictive capacity of TM in tumor invasiveness.
Results
The data revealed that positive immunoreactivity for TM was inversely correlated with clinical stage and DNA methyltransferase 1 immunoreactivity. Decreased TM expression could predict the aggressive tumor growth and advanced clinical stage in bladder cancer. When TM was inhibited, tumor growth rate and invasion ability were augmented in vitro and in vivo. The underlying changes included increased cell proliferation, enhanced epithelial-mesenchymal transition (EMT) and angiogenesis. Moreover, inhibition of NF-κB activation significantly increased TM expression and attenuated tumor aggressiveness in bladder cancer.
Conclusions
TM plays an important role in bladder cancer tumor aggressiveness in vitro and in vivo and is a clinically significant predictor that may represent a suitable therapeutic target for bladder cancer.
... CAPE kills various types of cancer cells but is innocuous to normal cells. There are several studies reporting the in vitro and in vivo inhibitory effects of CAPE in multiple cancer models, such as colon cancer [27], lung cancer [28], melanoma [29], glioma [30], pancreatic cancer [31], gastric cancer [32], cholangiocarcinoma [33], hepatocellular carcinoma [34], and our data in breast cancer3536373839. The role of CAPE as an inhibitor of angiogenesis, tumor metastasis and invasion has been demonstrated in several models of angiogenesis, both in vitro [40] and in vivo in colon cancer cells [41]. ...
... MDA-231 cells are known to secrete vascular endothelial growth factor (VEGF) necessary for endothelial cell growth and important in tumor blood vessel formation (MCF-7 cells secrete very little VEGF; not shown). CAPE, from 0 to 40 µM, inhibits VEGF secretion by MDA-231 cells in a dosedependent fashion from 0 to 40 µM CAPE, with >90% suppression at the highest dose (Figure 5b).Figure 6,Table I) CAPE modulates NF-κB, it inhibits NF-κB binding activity and decreases the expression of NF-κB1 (p50) and RelA (p65) [21, 22, 32, 33]. We show here that CAPE inhibits NF-κB activity in MDA-231 cells (Figure 6a). ...
Breast cancer (BC) patients use alternative and natural remedies more than patients with other malignancies. Specifically, 63-83% use at least one type of alternative medicine and 25-63% use herbals and vitamins. Propolis is a naturopathic honeybee product, and CAPE (caffeic acid phenethyl ester), is a major medicinal component of propolis. CAPE, in a concentration dependent fashion, inhibits MCF-7 (hormone receptor positive, HR+) and MDA-231 (a model of triple negative BC (TNBC) tumor growth, both in vitro and in vivo without much effect on normal mammary cells and strongly influences gene and protein expression. It induces cell cycle arrest, apoptosis and reduces expression of growth and transcription factors, including NF-κB. Notably, CAPE down-regulates mdr-1 gene, considered responsible for the resistance of cancer cells to chemotherapeutic agents. Further, CAPE dose-dependently suppresses VEGF formation by MDA-231 cells and formation of capillary-like tubes by endothelial cells, implicating inhibitory effects on angiogenesis. In conclusion, our results strongly suggest that CAPE inhibits MDA-231 and MCF-7 human breast cancer growth via its apoptotic effects, and modulation of NF-κB, the cell cycle, and angiogenesis.
... They concluded that several plants had bactericidal and/or anti-adhesive effects on H. pylori, and therefore, ingestion of these plants could provide a potent alternative therapy for H. pylori infection which overcomes the problem of resistance associated with current antibiotic treatment. Honey is a traditional remedy for dyspepsia and there have been a number of reports suggesting that honey can inhibit the growth of H. pylori.56,57 Abdel-Latif et al.56 and Wu et al.57 reported that CAPE (caffeic acid phenethyl ester), an active component of propolis from honeybee hives, has anti-inflammatory, anti-carcinogenic and antibacterial properties, can effectively reduce the inflammatory triggers after H. pylori infection.In addition to fruit and vegetables, vitamins have been studied, since oxidative stress plays a critical role in the augmented mucosal damage in H. pylori infection. ...
... Honey is a traditional remedy for dyspepsia and there have been a number of reports suggesting that honey can inhibit the growth of H. pylori.56,57 Abdel-Latif et al.56 and Wu et al.57 reported that CAPE (caffeic acid phenethyl ester), an active component of propolis from honeybee hives, has anti-inflammatory, anti-carcinogenic and antibacterial properties, can effectively reduce the inflammatory triggers after H. pylori infection.In addition to fruit and vegetables, vitamins have been studied, since oxidative stress plays a critical role in the augmented mucosal damage in H. pylori infection. An antioxidant, α-tocopherol (vitamin E), could reduce the aggravation of stress-associated gastric mucosal damage.58 ...
Helicobacter pylori (H. pylori) infection causes peptic ulcer disease, mucosa-associated lymphoid tissue (MALT) lymphomas and gastric adenocarcinomas, for which the pathogenesis of chronic gastric inflammation prevails and provides the pathogenic basis. Since the role of H. pylori infection is promoting carcinogenesis rather than acting as a direct carcinogen, as several publications show, eradication alone cannot be the right answer for preventing H. pylori-associated gastric cancer. Therefore, a non-antimicrobial approach has been suggested to attain microbe-associated cancer prevention through controlling H. pylori-related chronic inflammatory processes and mediators responsible for carcinogenesis. Phytoceutical is a term for plant products that are active on biological systems. Phytoceuticals such as Korean red ginseng, green tea, red wine, flavonoids, broccoli sprouts, garlic, probiotics and flavonoids are known to inhibit H. pylori colonization, decrease gastric inflammation by inhibiting cytokine and chemokine release, and repress precancerous changes by inhibiting nuclear factor-kappa B DNA binding, inducing profuse levels of apoptosis and inhibiting mutagenesis. Even though further unsolved issues are awaited before phytoceuticals are accepted as a standard treatment for H. pylori infection, phytoceuticals can be a mighty weapon for either suppressing or modulating the disease-associated footprints of H. pylori infection.
... A large rise in reactions may be reversed by treatment with caffeic acid. Tumor development and spread may be inhibited by caffeic acid as well [39]. In addition, two investigations have indicated that quercetin protects mucosal tissue against ulcer-induced damage. ...
Gastric cancer is one of the most common cancers of the gastrointestinal tract. Although surgery is the primary treatment, serious maladies that dissipate to other parts of the body may require chemotherapy. As there is no effective procedure to treat stomach cancer, natural small molecules are a current focus of research interest for the development of better therapeutics. Chemo-therapy is usually used as a last resort for people with advanced stomach cancer. Anti-colon cancer chemotherapy has become increasingly effective due to drug resistance and sensitivity across a wide spectrum of drugs. Naturally-occurring substances have been widely acknowledged as an important project for discovering innovative medications, and many therapeutic pharmaceuticals are made from natural small molecules. Although the beneficial effects of natural products are as yet unknown, emerging data suggest that several natural small molecules could suppress the progression of stomach cancer. Therefore, the underlying mechanism of natural small molecules for pathways that are directly involved in the pathogenesis of cancerous diseases is reviewed in this article. Chemotherapy and molecularly-targeted drugs can provide hope to colon cancer patients. New discoveries could help in the fight against cancer, and future stomach cancer therapies will probably include molecularly formulated drugs.
... CAPE is the major bioactive molecule of propolis, which is obtained from honeybee products. Multiple studies have reported the in vitro and in vivo anticancer potential of CAPE against various cancers including lung cancer, colon cancer, glioma, pancreatic cancer, breast cancer, hepatocellular carcinoma, gastric cancer, cholangiocarcinoma, and melanoma [107][108][109][172][173][174][175][176]. A study reported the anticancer, anti-inflammatory, and immunomodulatory effects of CAPE in vitro [177]. ...
Human skin is continuously subjected to environmental stresses, as well as extrinsic and intrinsic noxious agents. Although skin adopts various molecular mechanisms to maintain homeostasis, excessive and repeated stresses can overwhelm these systems, leading to serious cutaneous damage, including both melanoma and non-melanoma skin cancers. Phytochemicals present in the diet possess the desirable effects of protecting the skin from damaging free radicals as well as other benefits. Dietary phytochemicals appear to be effective in preventing skin cancer and are inexpensive, widely available, and well tolerated. Multiple in vitro and in vivo studies have demonstrated the significant anti-inflammatory, antioxidant, and anti-angiogenic characteristics of dietary phytochemicals against skin malignancy. Moreover, dietary phytochemicals affect multiple important cellular processes including cell cycle, angiogenesis, and metastasis to control skin cancer progression. Herein, we discuss the advantages of key dietary phytochemicals in whole fruits and vegetables, their bioavailability, and underlying molecular mechanisms for preventing skin cancer. Current challenges and future prospects for research are also reviewed. To date, most of the chemoprevention investigations have been conducted preclinically, and additional clinical trials are required to conform and validate the preclinical results in humans.
... CAPE is the major bioactive molecule of propolis, which is obtained from honeybee products. Multiple studies have reported the in vitro and in vivo anticancer potential of CAPE against various cancers including lung cancer, colon cancer, glioma, pancreatic cancer, breast cancer, hepatocellular carcinoma, gastric cancer, cholangiocarcinoma, and melanoma [107][108][109][172][173][174][175][176]. A study reported the anticancer, anti-inflammatory, and immunomodulatory effects of CAPE in vitro [177]. ...
... Caffeic acid phenethyl ester (CAPE), a catechol destroys different cancer cell types but is harmless to normal cells. Numerous studies have shown that CAPE's inhibitory effects occur both in vitro and in vivo in many cancer models such as colon cancer [205], lung cancer [206], melanoma [207], glioma [208], pancreatic cancer [209], gastric cancer [210], cholangiocarcinoma [211], and hepatocellular carcinoma [212]. ...
The world population is aging, and cancer is always considered to be one of the major causes of death all over the globe. The advent of recent drug-targeted therapies undoubtedly is going to reduce the incidence of cancer over the coming years. However, the frequency of occurrence of such chronic diseases like cancer would continue to increase. Therefore, the search for a safer and cost-effective treatment is urgently needed. Phytochemicals found in plants, foods, vegetables, tea, etc. have emerged as proven therapeutic compounds modulating signaling pathways involved in cancer. We carried out a structured search of bibliographic databases for peer-reviewed research literature using the keywords: cancer chemoprevention, flavonoids, dietary polyphenols, terpenoids, bioactive, microbiota. Quality of the retrieved papers and characteristic outcomes of the articles included in the study was assessed by employing standard tools and deductive qualitative content analysis methodology. The development of personalized supplements comprising particular phytochemicals has been the key, especially dealing with chronic inflammatory disorders like cancer. Better understanding at the molecular level explains the influence of phytochemicals on human health, which has been extensively covered through this review. Moreover, the wide collection of dietary polyphenols that has significant properties in reference to human health has been highlighted. Furthermore, the etiology of end products of such phytochemicals, especially on the modulation of gut microbiota and the host-microbial interactions thereof, need to be properly understood. The present study summarizes the chemoprevention and treatment of cancer using the bioactive components, including flavonoids, dietary polyphenols, and terpenoids. Likewise, the effect of dietary polyphenols on the human gut microbiota has been realized more recently. However, more research is needed in this field, especially focused on the communications, interlinks between the gut microbiota and polyphenols with the precise mechanism of action.
... Tumor aggressiveness in gastric cancer was shown by investigating 90 human cancer tissues versus 50 nonmalignant specimens. A higher NF-jB expression in cancer tissue versus normal mucosa (31% vs. 4%, p < 0.0001) was found along with activation of metalloproteinase 9 (MMP-9), IL-b1, and IL-8 in AGS cells [120]. Another example of NF-jB signaling and chronic inflammation was provided by Kwon et al. [121] who demonstrated that the Vitamin D(3) upregulated protein 1 (VDUP1) with its tumor suppressive effect was shown in VDUP1 knockout (KO) revealing that VDUP1 negatively regulates Helicobacter pylori (H. ...
Transcription factors (TFs) are proteins that control the transcription of genetic information from DNA to mRNA by binding to specific DNA sequences either on their own or with other proteins as a complex. TFs thus support or suppress the recruitment of the corresponding RNA polymerase. In general, TFs are classified by structure or function. The TF, Nuclear factor kappa-light-chain-enhancer of activated B cells (NF-jB), is expressed in all cell types and tissues. NF-jB signaling and crosstalk are involved in several steps of carcinogenesis including in sequences involving pathogenic stimulus, chronic inflammation, fibrosis, establishment of its remodeling to the precancerous niche (PCN) and transition of a normal cell to a cancer cell. Triggered by various inflammatory cytokines, NF-jB is activated along with other TFs with subsequent stimulation of cell proliferation and inhibition of apoptosis. The involvement of NF-jB in carcinogenesis provides an opportunity to develop anti-NF-jB therapies. The complexity of these interactions requires that we elucidate those aspects of NF-jB interactions that play a role in carcinogenesis, the sequence of events leading to cancer.
... Caffeic acid phenethyl ester (CAPE) is a major medicinal component of propolis, which is derived from honeybee products. There are several studies reporting the inhibitory effects of CAPE on many cancer types both in vitro and in vivo, including colon cancer, lung cancer, melanoma, glioma, pancreatic cancer, gastric cancer, cholangiocarcinoma, hepatocellular carcinoma, and breast cancer [18][19][20][21][22][23][24][25][26]. CAPE was shown to exhibit anti-mitogenic, anti-carcinogenic, anti-inflammatory, and immunomodulatory properties in vitro [27]. ...
Skin is the largest human organ, our protection against various environmental assaults and noxious agents. Accumulation of these stress events may lead to the formation of skin cancers, including both melanoma and non-melanoma skin cancers. Although modern targeted therapies have ameliorated the management of cutaneous malignancies, a safer, more affordable, and more effective strategy for chemoprevention and treatment is clearly needed for the improvement of skin cancer care. Phytochemicals are biologically active compounds derived from plants and herbal products. These agents appear to be beneficial in the battle against cancer as they exert anti-carcinogenic effects and are widely available, highly tolerated, and cost-effective. Evidence has indicated that the anti-carcinogenic properties of phytochemicals are due to their anti-oxidative, anti-inflammatory, anti-proliferative, and anti-angiogenic effects. In this review, we discuss the preventive potential, therapeutic effects, bioavailability, and structure–activity relationship of these selected phytochemicals for the management of skin cancers. The knowledge compiled here will provide clues for future investigations on novel oncostatic phytochemicals and additional anti-skin cancer mechanisms.
... In vitro research studies have clearly shown the cytotoxic properties of CAPE against: cells of pulmonary carcinoma, gastric carcinoma, colorectal carcinoma, malignant melanoma, hepatic carcinoma, pancreatic carcinoma, as well as cervical carcinoma [26][27][28][29][30][31][32][33]. ...
One of the deadliest cancers among women is a breast cancer. Research has shown that two natural substances occurring in propolis, caffeic acid (CA) and caffeic acid phenethyl ester (CAPE), have significant anticancer effects. The purpose of our in vitro study was to compare cytotoxic activity and migration rate inhibition using CA and CAPE (doses of 50 and 100 µm) against triple-negative, MDA-MB-231 breast adenocarcinoma line cells, drawn from Caucasian women. Viability was measured by XTT-NR-SRB assay (Tetrazolium hydroxide-Neutral Red-Sulforhodamine B) for 24 h and 48 h periods. Cell migration for wound healing assay was taken for 0 h, 8 h, 16 h, and 24 h periods. CAPE displayed more than two times higher cytotoxicity against MDA-MB-231 cells. IC50 values for the XTT assay were as follows: CA for 24 h and 48 h were 150.94 µM and 108.42 µM, respectively, while CAPE was 68.82 µM for 24 h and 55.79 µM for 48 h. For the NR assay: CA was 135.85 µM at 24 h and 103.23 µM at 48 h, while CAPE was 64.04 µM at 24 h and 53.25 µM at 48 h. For the SRB assay: CA at 24 h was 139.80 µM and at 48 h 103.98 µM, while CAPE was 66.86 µM at 24 h and 47.73 µM at 48 h. Both agents suspended the migration rate; however, CAPE displayed better activity. Notably, for the 100 µM CAPE dose, motility of the tested breast carcinoma cells was halted.
... Numerous research studies have demonstrated that CAPE possesses versatile pharmacological properties such as antibacterial, anti-inflammatory, antiproliferative, antiviral, anticarcinogenic, antidiabetic, antileukemic, chemopreventive and neuroprotective effects (Murtaza et al., 2014;Tolba et al., 2013). Recent preclinical studies have shown that CAPE was efficient in inhibiting different kinds of cancers such as colorectal cancer, lung cancer, breast cancer, pancreatic cancer, gastric cancer, melanoma, glioma, cholangiocarcinoma and hepatocellular carcinoma in micromolar concentrations (Chen et al., 2004(Chen et al., , 2008Kuo et al., 2006;Lee et al., 2008;Onori et al., 2009;Wu et al., 2007Wu et al., , 2011Xiang et al., 2006). CAPE ester analogs including caffeic acid (4-hydroxylphenethyl) ester (CAHPE), caffeic acid (3,4-dihydroxyphenethyl) ester (CADPE) and a series of amide analogs including caffeic acid phenethyl amide (CAPA), caffeoyl tyramine (CATA) and caffeoyl dopamine (CADA) are all potent antioxidants Son and Lewis, 2002). ...
Caffeic acid has been widely recognized as a versatile pharmacophore for synthesis of new chemical entities, among which caffeic acid derived phenethyl esters and amides are the most extensively-investigated bioactive compounds with potential therapeutical applications. However, the natural biosynthetic routes for caffeic acid derived phenethyl esters or amides remain enigmatic, limiting their bio-based production. Herein, product-directed design of biosynthetic schemes allowed the development of thermodynamically favorable pathways for these compounds via acyltransferase (ATF) mediated trans-esterification. Production based screening identified a microbial O-ATF from Saccharomyces cerevisiae and a plant N-ATF from Capsicum annuum capable of forming caffeic acid derived esters and amides, respectively. Subsequent combinatorial incorporation of caffeic acid with various aromatic alcohol or amine biosynthetic pathways permitted the de novo bacterial production of a panel of caffeic acid derived phenethyl esters or amides in Escherichia coli for the first time. Particularly, host strain engineering via systematic knocking out endogenous caffeoyl-CoA degrading thioesterase and pathway optimization via titrating co-substrates enabled production enhancement of five caffeic acid derived phenethyl esters and amides, with titers ranging from 9.2 to 369.1mg/L. This platform expanded the capabilities of bacterial production of high-value natural aromatic esters and amides from renewable carbon source via tailoring non-natural biosynthetic pathways.
... Therefore, the inhibition of these molecules by CAPE could result in suppression of many genes during H. pyloriinduced inflammation. Wu et al [75] demonstrated that the activity of NFκB and the expression of matrix metalloproteinase9, IL1beta, and IL8 in gastric cancer cells by H. pylori significantly reversed by CAPE treatment, which suggested that CAPE could be promising adjuvant agent against gastric cancer. In Monogolian gerbils, CAPE treatment elicited antiinflammatory effects on H. pyloriinduced chronic gastritis. ...
... CAPE-and Artepillin C (ARC)-rich propolis extracts were suggested as natural anti-PAK1 inhibitors and NF1/NF2 remedies [24,25]. CAPE was shown to inhibit TPA (12-O-tetradecanoylphorbol-13-acetate)-induced tumors [26,27], and suggested as an anticancer drug for treatment of colorectal carcinoma [28], hepatoma [29] and gastric carcinoma [30]. CAPE-induced cell cycle arrest and apoptosis are mediated by down-regulation of beta-catenin [31,32] and p38MAPK [33] signaling. ...
Caffeic Acid Phenethyl Ester (CAPE) is a key component in New Zealand propolis, known for a variety of health promoting and therapeutic potentials. We investigated the molecular mechanism of anticancer and anti-metastasis activities of CAPE. cDNA array performed on the control and CAPE-treated breast cancer cells revealed activation of DNA damage signaling involving upregulation of GADD45α and p53 tumor suppressor proteins. Molecular docking analysis revealed that CAPE is capable of disrupting mortalin-p53 complexes. We provide experimental evidence and demonstrate that CAPE induced disruption of mortalin-p53 complexes led to nuclear translocation and activation of p53 resulting in growth arrest in cancer cells. Furthermore, CAPE-treated cells exhibited downregulation of mortalin and several other key regulators of cell migration accountable for its anti-metastasis activity. Of note, we found that whereas CAPE was unstable in the culture medium (as it gets degraded into caffeic acid by secreted esterases), its complex with gamma cyclodextrin (γCD) showed high efficacy in anti-tumor and anti-metastasis assays in vitro and in vivo (when administered through either intraperitoneal or oral route). The data proposes that CAPE-γCD complex is a potent anti-cancer and anti-metastasis reagent.
... Ogura et al. reported that the degree of gastritis induced by a mutant strain of H. pylori lacking capacity for NF-κB activation is lower than that with wild-type infection in gerbils [72]. Therefore, it is considered that inhibition of NF-κB could be a target for the prevention of H. pylori-associated gastric cancer [73]. Caffeic acid phenethyl ester (CAPE), a naturally-derived NF-κB inhibitor, was analyzed on H. pylori-induced chronic gastritis using the gerbil model [74]. ...
Since the discovery of Helicobacter pylori (H. pylori), many efforts have been made to establish animal models for the investigation of the pathological features and molecular mechanisms of gastric carcinogenesis. Among the animal models, Mongolian gerbils and mice are particularly useful for the analysis of H. pylori-associated inflammatory reactions and gastric cancer development. Inhibitors of oxidative stress, cyclooxygenase-2 (COX-2) and nuclear factor-κB, exert preventive effects on chronic gastritis and the development of adenocarcinomas in H. pylori-infected gerbils. Genetically-modified mouse models, including transgenic and knockout mice, have also revealed the importance of p53, COX-2/prostaglandin, Wnt/β-catenin, proinflammatory cytokines, gastrin and type III mucin in the molecular mechanisms of gastric carcinogenesis. Microarray technology is available for comprehensive gene analysis in the gastric mucosa of mouse models, and epigenetics, such as DNA methylation, could be an alternative approach to correlate the observations in animal models with the etiology in humans.
... Gastric cancer is the most common cause of death from cancer and it is resistant to drug therapy. Oncogenes control cell proliferation, invasion and angiogenesis by the effect of matrix molecules and angiogenetic factors (Abdel et al. 2005, Wu et al. 2007, Chan et al. 2013. Therefore, we investigated the effects of CAPE as an anticancer agent and measured its effect on the matrix and angiogenesis. ...
Migration, invasion, metastasis and angiogenesis associated with cancer depend on the surrounding microenvironment. Angiogenesis, the growth of new capillaries, is a regulator of cancer growth and a useful target for cancer therapy. We examined matrix protein interactions in a gastric cancer cell culture that was treated with different doses of caffeic acid (3,4-dihydroxycinnamic acid) phenethyl ester (CAPE). We also investigated the relations among the levels of vascular endothelial growth factor (VEGF), matrix metalloproteinase-9 (MMP-9), endostatin (ES) and trombospondin-1 (TSP-1). Cytotoxity of CAPE was measured using the 3-(4,5-dmethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. We examined the behavior of cells on laminin and collagen I coated surfaces in response to the angiogenic effect of these matrix molecules. We examined the protein alterations of these matrix molecules immunohistochemically and measured the levels of VEGF, MMP-9, ES and TSP-1 using the ELISA test. We showed that application of CAPE to the gastric cancer cell line on tissue culture plastic, laminin and collagen I significantly decreased the VEGF and MMP-9 protein levels. We found that TSP-1 levels were increased significantly in the gastric cancer cells after application of CAPE. The protein levels of gastric cancer cells also were increased significantly when tissue was cultured on laminin and collagen I. Application of CAPE to cells on laminin or collagen I coated surfaces significantly increased all of the proteins except ES. ES levels were increased on the collagen I covered surfaces, but the laminin surface decreased the levels of ES significantly. We demonstrated the beneficial effect of CAPE on a gastric cancer cell line including inhibition of proliferation and induction of some proteins that might be related to decreased angiogenesis.
... Propolis has caffeic acid phenethyl ester (CAPE), and it inhibits MCF-7 (hormone receptor positive, HR+) and MDA-231 (a model of triple-negative breast cancer [TNBC] tumor growth, both in vitro and in vivo without much effect on normal mammary cells and strongly influences gene and protein expression. [56,57] Similarly, CAPE has inhibitory effects on multiple cancer models, such as colon cancer, [58,59] lung cancer, [53,60] melanoma, [61] glioma, [62] pancreatic cancer, [63] gastric cancer, [64] cholangiocarcinoma, [65] hepatocellular carcinoma, [66] prostate cancer, breast cancer, [67] and human renal cell carcinoma. [68] The role of CAPE as an inhibitor of angiogenesis, tumor metastasis, and invasion has been demonstrated in several models of angiogenesis, both in vitro [58,69] and in vivo. ...
The natural products are gaining immense importance in the domain of nutrition to prevent various maladies and improve the quality of life. Among these, natural exudates are of significant worth as these biochemical compounds are released by various living entities having pharmacological properties for utilization in various drug developments. These natural exudates are the promising source for the discovery of new medications. Numerous bioactive moieties collected by honeybees from exudates and buds of particular trees and plants, considered to be utilized as defensive barrier with special reference to propolis. It generally contains numerous biochemical components, i.e., polyphenols, steroids, terpenoids, and amino acids. They also contain isoferulic acid, sinapinic acid, caffeic acid, and chrysin responsible for antibacterial perspectives. With special attention to propolis, it has been utilized in folk medicines due to several of its therapeutic activities, i.e., antioxidant, antidiabetic, anti-inflammatory, antiviral, and anticancer properties. In this context, it is extensively used in foodstuffs and beverages to improve health related disorders like inflammation, diabetes, heart disease, protects injured gums, and cancer insurgence. Moreover, it has been used to curtail stomatology, gastroenterology, skin lesions, and otorhinolaryngologic and respiration diseases.
... In vitro studies reveal the cytotoxic properties of CAPE against the cell lines of colorectal carcinoma [26,27], pulmonary carcinoma [28], malignant melanoma [29], gastric carcinoma [30], pancreatic carcinoma [31], hepatic carcinoma [32], cervical carcinoma [33] cholangiocarcinoma [34], glioma [35] and some other cell lines of breast cancer [36,37]. ...
Chemotherapy of breast cancer could be improved by bioactive natural substances, which may potentially sensitize the carcinoma cells' susceptibility to drugs. Numerous phytochemicals, including propolis, have been reported to interfere with the viability of carcinoma cells. We evaluated the in vitro cytotoxic activity of ethanol extract of propolis (EEP) and its derivative caffeic acid phenethyl ester (CAPE) towards two triple-negative breast cancer (TNBC) cell lines, MDA-MB-231 and Hs578T, by implementation of the MTT and lactate dehydrogenase (LDH) assays. The morphological changes of breast carcinoma cells were observed following exposure to EEP and CAPE. The IC50 of EEP was 48.35 µg∙mL-1 for MDA-MB-23 cells and 33.68 µg∙mL-1 for Hs578T cells, whereas the CAPE IC50 was 14.08 µM and 8.01 µM for the MDA-MB-231 and Hs578T cell line, respectively. Here, we report that propolis and CAPE inhibited the growth of the MDA-MB-231 and Hs578T lines in a dose-dependent and exposure time-dependent manner. EEP showed less cytotoxic activity against both types of TNBC cells. EEP and, particularly, CAPE may markedly affect the viability of breast cancer cells, suggesting the potential role of bioactive compounds in chemoprevention/chemotherapy by potentiating the action of standard anti-cancer drugs.
... CAPE possesses a number of important biological activities, including anti-bacterial, antiviral, anti-fungal, anti-oxidant, anti-inflammatory, and anti-cancer properties [10][11][12][13][14][15][16]. CAPE has been shown to be cytotoxic to many types of cancer cells, including breast cancer cells, while having no such effects against normal cells [17][18][19][20][21][22][23][24][25][26]. We have previously demonstrated that CAPE: (i) inhibits the growth of MDA-MB-231 (MDA-231) triple-negative (ER−, PR−, Her2-) BC cells (TNBC), and MCF-7 (ER+/PR+) BC cells both in vivo and in vitro; (ii) inhibits growth of breast cancer stem cells; (iii) induces cell cycle arrest and apoptosis and (iv) suppresses angiogenesis [25,26]. ...
Alternative remedies for cancer treatment is a multi-billion dollar industry. In particular, breast cancer (BC) patients use alternative and natural remedies more frequently than patients with other malignancies. Propolis is an example of a honeybee-produced naturopathic formulation, contents of which differ by geographic location. It is readily available, affordable, and in use safely since ancient times globally. Caffeic acid phenethyl ester (CAPE) is a major active component in propolis and is thought to be responsible for its varied properties, including antibacterial, antiviral, antifungal, antioxidant, anti-inflammatory and anticancer. CAPE is effective in many models of human cancer, including BC as we have previously shown. CAPE affects genes associated with tumor cell growth and survival, angiogenesis and chemoresistance. We demonstrate that these are related in part to CAPE's role as a histone deacetylase inhibitor, a class of drugs designated as epigenetic agents that modulate the activities of oncogenes and tumor suppressor genes. CAPE and propolis, cause an accumulation of acetylated histone proteins in MCF-7 (ER+) and MDA-MB-231 (ER-/PR-/Her2-) cells with associated decreases in ER and PR in MCF-7 cells, and upregulation of ER and decrease in EGFR in MDA-231 cells. In addition, these products reduced activated phosphorylated Her2 protein in SKBR3 (Her2 +) cells. Interestingly, propolis, when normalized for CAPE content, appears to be more potent than CAPE alone similarly to the greater effects of complete foods than isolated components. These data provide a potential mechanistic basis for one of the oldest naturopathic agents used in medicine and cancer treatment.
... propolis that possesses a plethora of biological activities. Several studies reported its broad-spectrum anticancer activity in vitro and in vivo in multiple cancer models, as colon cancer [11], lung cancer [12], melanoma [13], glioma [14], pancreatic cancer [15], gastric cancer [16], cholangiocarcinoma [17], hepatocellular carcinoma [18], breast cancer [19] and recently, prostate cancer [20]. Several reports indicated that CAPE has preferential cytotoxicity on tumor cells rather than normal cells [21][22]. ...
Docetaxel (DOC) and paclitaxel (PTX) play an important role in the
... Nuclear expression of NF-κB was observed much more in gastric cancer tissue than in nonmalignant gastric tissues. The activity of NF-κB and the expressions of MMP-9, IL-1β, and IL-8 in gastric adenocarcinoma epithelial cell line were activated by H. Pylori infection, and the augmented responses could be significantly reversed by CAPE treatment (27). ...
Cancer prevention and treatment strategies have attracted increasing interest. Caffeic acid phenethyl ester (CAPE), an active component of propolis extract, specifically inhibits NF-κB at μM concentrations and shows ability to stop 5-lipoxygenase-catalyzed oxygenation of linoleic acid and arachidonic acid. Previous studies have demonstrated that CAPE exhibits antioxidant, antiinflammatory, antiproliferative, cytostatic, antiviral, antibacterial, antifungal, and, most improtantly, antineoplastic properties. The primary goal of the present review is to summarize and critically evaluate the current knowledge regarding the anticancer effect of CAPE in different cancer types.
... CAPE has an antioxidant effect (25,26). Moreover, anti-inflammatory, anticarcinogenic, neuroprotective, and antiepileptic effects have also been reported (12,27,28). It was suggested that using CAPE in combination with anticancer drugs was protective against anticancer toxicity such as caused by MTX and doxorubicin (7,25,29). ...
ZET Amaç: Kanser hastalar›nda metotreksata ba¤l› nörotoksisite önemli klinik bir problemdir. Fakat metotreksat›n (MTX) neden oldu¤u nörotoksisitenin mekanizmas› tam olarak bilinmemektedir. Bu çal›flman›n amaçlar›; MTX'e ba¤l› nörotoksisitenin patogenezinde ma-londialdehid (MDA), süperoksid dismutaz (SOD), glutatyon peroksidaz (GSH-Px) ve katalaz (CAT)'›n olas› rolü ile ratlar›n siyatik sinir, beyin sap› ve medulla spinalisinde MTX'e ba¤l› nörotoksisitesinde koruyucu etkisi olup olmad›¤›n› araflt›rmakt›r. Gereç ve Yöntem: Toplam 19 adet Wistar erkek rat üç deney grubuna ayr›ld›. Grup 1: Kontrol grubu, Grup 2: MTX alan grup, Grup 3: MTX ve kafeik asit fenetil ester (CAPE) alan grup. MTX ve MTX + CAPE gruplar›na deneyin ikinci gününde MTX 20 mg/kg tek doz periton içine verildi. MTX CAPE grubuna CAPE 10 µmol/kg/gün intraperitoneal olarak 7 gün verildi. Bulgular: MTX grubunda siyatik sinir ve spinal kord dokusunda kontrol grubu ile karfl›laflt›r›ld›¤›nda CAT ve GSH-Px aktivitelerinde ar-t›fl bulundu. MTX + CAPE ile MTX grubu karfl›laflt›r›ld›¤›nda CAT ve GSH-Px aktivitelerinde azalma saptand›. MTX grupta spinal kord ve beyin sap› dokular›nda SOD aktivitesi kontrolle karfl›laflt›r›ld›¤›nda azalma saptan›rken, siyatik sinirde anlaml› fark bulunmad›. Spi-nal kord ve beyin sap› dokular›nda SOD aktivitesi MTX + CAPE grubunda MTX grupla karfl›laflt›r›ld›¤›nda anlaml› derecede art›fl bulun-du. MDA seviyesi MTX grupta kontrol grubuna göre istatistiksel olarak anlaml› derecede yüksekti. MTX + CAPE grubunda MDA sevi-yeleri MTX grubuna göre istatistiksel olarak anlaml› derecede düflük bulundu. Yorum: Bu sonuçlar MTX'in rat siyatik sinir, medulla spinalis ve beyin sap›nda oksidatif stresi art›rd›¤›n› ve CAPE'nin antioksidan etki-si nedeniyle oksidatif strese karfl› koruyucu etkisini gösterir. Anahtar Kelimeler: Kafeik asit fenetil ester, metotreksat, oksidatif stres, siyatik sinir, medulla spinalis.
... Research indicates that these polyphenols may have antioxidant characteristics with potential health benefits including reducing the risk of cancer (11). We have shown that caffeic acid, a polyphenol extracted from the propolis of honeybee hives, exerts antiproliferative effects on cholangiocarcinoma (8) and other tumortypes (12)(13)(14) in vitro and in vivo by inhibiting the nuclear factor-κ-B pathway (8). Furthermore, high concentrations of resveratrol have been shown to be antiproliferative in a cholangiocarcinoma cell line (5) as well as in other tumor types (15)(16)(17)(18)(19) through a number of different mechanisms including increased Cyclooxygenase 2 expression (15), activation of Forkhead proteins (16), facilitation of death receptor complex formation (17,18) or cell cycle arrest (19). ...
Cholangiocarcinomas are devastating cancers that are resistant to chemotherapies. Resveratrol, a food-derived polyphenol with antitumorigenic properties, can regulate the expression of cytochrome p450 1b1 (Cyp1b1), which may confer chemoresistance in various cancers. Our aims were to assess the effects of resveratrol on the sensitivity of cholangiocarcinoma cells to chemotherapeutic agents and show an association between Cyp1b1 expression and chemosensitivity. Cholangiocarcinoma cell lines were treated with resveratrol before the addition of 5-fluorouracil (5-FU), gemcitabine, or mitomycin C. Cell proliferation and apoptosis were assessed by MTS assays and Annexin staining. Resveratrol effects on cholangiocarcinoma tumor sensitivity to 5-FU was assessed in an in vivo xenograft model using Mz-ChA-1 cells. After resveratrol treatment, Cyp1b1 expression was assessed by real-time PCR and immunoblotting. Stable-transfected cell lines with Cyp1b1 expression knocked down (Mz-Cyp1b1) were used to assess sensitivity to chemotherapeutic agents by MTS assays and Annexin staining and in a xenograft model using Mz-ChA-1 and Mz-Cyp1b1 cells, respectively. For each chemotherapeutic agent, co-treatment with resveratrol in vitro decreased cell proliferation and increased apoptosis to a greater extent than with the chemotherapeutic agent alone. In vivo, 5-FU+resveratrol decreased tumor size and increased TUNEL staining to a greater extent than 5-FU alone. In parallel, resveratrol decreased Cyp1b1 expression in Mz-ChA-1 cells and in cholangiocarcinoma tumors. Mz-Cyp1b1 cells were more sensitive to chemotherapeutic agents in vitro than mock-transfected cells, and Mz-Cyp1b1-induced tumors were more susceptible to 5-FU treatment. We suggest that resveratrol treatment may be a useful adjunct therapy to improve chemosensitivity in cholangiocarcinoma.
... The plant polyphenolic compound CAPE exerts a number of actions which are relevant for potential therapeutic applications in the digestive tract. These include protective effects against experimentally-induced intestinal damage and inflammation (Koltuksuz et al., 1999;Fitzpatrick et al., 2001;Ara et al., 2006;Grémy et al., 2006;Ek et al., 2007) and chemopreventive actions against stomach and colon cancer (Mahmoud et al., 2000;Borrelli et al., 2002;Ribeiro and Safatle-Ribeiro, 2007;Wu et al., 2007). Here, we have shown that CAPE produced a direct inhibitory effect on the motility of the isolated rat ileum. ...
Caffeic acid phenethyl ester (CAPE) exerts pharmacological actions (e.g. anti-inflammatory, chemopreventive) which are relevant for potential clinical application in the digestive tract. However, no study has been published on its possible effects on intestinal motility, to date. In the present study, we investigated the effect of this plant-derived polyphenolic compound on the spontaneous contractions of the rat isolated ileum. CAPE reduced (in a tetrodotoxin-insensitive manner) spontaneous ileal contractions and this effect was reduced by the L-type Ca2+ channel blocker nifedipine and the chelant of calcium ethylenediaminetetraacetic acid. However, the effect of CAPE was not modified by a number of inhibitors/antagonists such as of phentolamine plus propranolol, atropine, tetrodotoxin, cyclopiazonic acid, omega-conotoxin, apamin, NG-nitro-L-arginine methyl ester, 3-isobutyl-1-methylxanthine, 9-(tetrahydro-2-furanyl)-9H-purin-6-amine, 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one or a combination of SR 140333, SR48968 and SR142801. In conclusion our study shows that (i) CAPE relaxed myogenic contractions of rat ileum and that (ii) this effect occurs, at least in part, throughout a mechanism involving L-type Ca2+ channels.
... Research indicates that these polyphenols may have antioxidant characteristics with potential health benefits including reducing the risk of cancer (83). Caffeic acid, a polyphenol extracted from the propolis of honeybee hives, exerts antiproliferative effects on cholangiocarcinoma (71) and other tumor types (22,99) in vitro and in vivo by inhibiting the nuclear factor-B pathway (71). Furthermore, high concentrations of resveratrol have been shown to be antiproliferative in a cholangiocarcinoma cell line (80) as well as in other tumor types (10, 21, 87) through a number of different mechanisms including increased cyclooxygenase 2 expression (87), facilitation of death receptor complex formation (20,21), or cell cycle arrest (10). ...
Cholangiocarcinomas arise after the neoplastic transformation of the cholangiocytes that line the intra- and extrahepatic biliary epithelium. Symptoms usually do not present until late in the course of the disease, at which time they are relatively resistant to chemotherapeutic agents and as such are difficult to treat and display a poor prognosis. Because of the relative rarity of this disease, the overall volume of research into the molecular pathophysiology associated with this disease is small compared with other more prevalent tumors. However, the incidence of this devastating cancer is on the rise and renewed efforts to understand the pathogenesis of cholangiocarcinoma is needed to design novel therapeutic strategies to combat this disease. This review summarizes the recent advances into our knowledge and understanding of cholangiocarcinoma and highlights potential novel therapeutic strategies that may prove useful to treat this deadly disease.
... 22 Inhibition of NF-jB could be a promising target for prevention and adjuvant therapy of H. pylori-associated gastric disorders. 3,23 The Mongolian gerbil (Meriones unguiculatus) provides a useful animal model of H. pylori-induced chronic active gastritis, allowing investigation of morbidity-related epithelial alterations in the gastric mucosa and their development into gastric neoplasia. 24 We have previously demonstrated that some natural products in food such as a fruit-juice concentrate of Japanese apricot and nordihydroguaiaretic acid, an antioxidant to preserve food and oils, and canolol, a potent oxygen radicals scavenger contained in canola oil, have suppressive effects on H. pyloriinduced gastric disorders in Mongolian gerbils. ...
Nuclear factor-kappaB (NF-kappaB) plays a major role in host inflammatory responses and carcinogenesis and as such is an important drug target for adjuvant therapy. In this study, we examined the effect of caffeic acid phenethyl ester (CAPE), an NF-kappaB inhibitor, on Helicobacter pylori (H. pylori)-induced NF-kappaB activation in cell culture and chronic gastritis in Mongolian gerbils. In AGS gastric cancer cells, CAPE significantly inhibited H. pylori-stimulated NF-kappaB activation and mRNA expression of several inflammatory factors in a dose-dependent manner, and prevented degradation of IkappaB-alpha and phosphorylation of p65 subunit. To evaluate the effects of CAPE on H. pylori-induced gastritis, specific pathogen-free male, 6-week-old Mongolian gerbils were intragastrically inoculated with H. pylori, fed diets containing CAPE (0-0.1%) and sacrificed after 12 weeks. Infiltration of neutrophils and mononuclear cells and expression of NF-kappaB p50 subunit and phospho-IkappaB-alpha were significantly suppressed by 0.1% CAPE treatment in the antrum of H. pylori-infected gerbils. Labeling indices for 5'-bromo-2'-deoxyuridine both in the antrum and corpus and lengths of isolated pyloric glands were also markedly reduced at the highest dose, suggesting a preventive effect of CAPE on epithelial proliferation. Furthermore, in the pyloric mucosa, mRNA expression of inflammatory mediators including tumor necrosis factor-alpha, interferon-gamma, interleukin (IL)-2, IL-6, KC (IL-8 homologue), and inducible nitric oxide synthase was significantly reduced. These results suggest that CAPE has inhibitory effects on H. pylori-induced gastritis in Mongolian gerbils through the suppression of NF-kappaB activation, and may thus have potential for prevention and therapy of H. pylori-associated gastric disorders.
... Thus, inhibitor kappa kinase (IKK) beta, a component of the NF-κB pathway [56], physically interacts with and phosphorylatively inactivates and suppresses TSC1, which leads to activation of the mTOR pathway [57]. Finally, a preclinical study showed that the activity of NF-κB and the expression of certain of its downstream effectors was reduced in a human gastric adenocarcinoma cell line, along with tumor growth and capacity for invasion, following exposure to caffeic acid phenethyl ester [58]. This finding has pathogenetic significance regarding the role of the NF-κB pathway in GAC. ...
Preclinical studies using human gastric adenocarcinoma (GAC) cell lines have shown that the mammalian target of rapamycin (mTOR) inhibitor, rapamycin, can inhibit tumor growth and that the extracellular signal-regulated kinase (ERK) of the Ras/Raf kinase/ERK pathway is related to chemoresistance and apoptosis. We examined the state of activation of components of mTOR, Ras/Raf kinase/ERK, and nuclear factor (NF)-kappaB signal transduction pathways, as well as cell cycle protein analyte correlates in GAC cases. Formalin-fixed paraffin-embedded tissue microarray blocks containing samples from 210 cases of GAC were examined. Immunohistochemistry was utilized to detect the following antigens: S100P, upstream stimulator of ERK, and NF-kappaB pathways; phosphorylated (p)-mTOR (Ser 2448), p-ERK-1/2 (Thr 202/Tyr 204), and one of their common down-stream effectors, p-p70S6K(Thr 389); p-NF-kappaBp65(Ser 536); and cell cycle associated proteins, Ki-67, and S phase kinase-associated protein (Skp)2. Immunoreactivity (0 to 4+) of protein expression and compartmentalization were assessed by bright-field microscopy. The majority of cases showed positive (1+ to 4+) cytoplasmic/plasmalemmal p-mTOR (88%), and moderate-strong (2+ to 4+) nuclear p-p70S6K (93%) and nuclear S100P (81%) expression. A subset of cases exhibited moderate-strong nuclear p-ERK-1/2 (15%) and p-NF-kappaBp65 (36%) expression. The majority of cases showed concomitant moderate-strong (2+ to 4+) nuclear Ki-67 (71%) and Skp2 (68%). Nuclear expression levels of p-ERK-1/2 and p-NF-kappaBp65, of p-p70S6K and p-NF-kappaB, and of Ki-67 and Skp2, respectively, showed significant linear correlations in GAC (p <0.001). Additionally, there were statistically significant differences in the mean expression levels of p-ERK-1/2 and p-NF-kappaBp65 in diffuse vs intestinal types of GAC, with higher levels of both in the diffuse type ( p = 0.001 and p <0.0001, respectively). In summary, morphoproteomic analysis reveals constitutive activation of mTOR and to some extent, Ras/Raf kinase and NF-kappaB pathways in GAC, as evidenced by increased cytoplasmic p-mTOR, nuclear translocation of p-p70S6K and p-ERK-1/2 phosphorylated at putative sites of activation (Ser 2448, Thr 389, and Thr 202/Tyr 204, respectively), as well as correlative expression of cell cycle analytes, Ki-67, and Skp2. These results suggest that a prospective study is warranted to evaluate the use of morphoproteomic profiling of individual patients with GAC in order to design combinatorial treatment strategies that target the mTOR, Ras/Raf kinase/ERK, and/or NF-kappaB pathways.
... [30][31][32] Its cancer chemopreventive effects occurs through the inhibition of COX-2 expression and the restoration of disrupted Gap junction intercellular communication in H-rastransformed, which may be associated in part with the blockade of NF-kB activation. 33,34 The current issue of the J Clin Gastroenterol features a paper by Wu et al, 35 from Taiwan, that address this topic on the role of H. pylori infection and the mechanisms that may lead to gastric cancer. The paper aims to clarify some of the molecular mechanisms [NF-kB, matrix metalloproteinase-9 (MMP-9), IL-1, and IL-8] involved in the chronic inflammation and carcinogenesis induced by H. pylori, as well as the effects of activated NF-kB inhibitor, CAPE. ...
Ginger (Zingiber officinale) is a spice and also an herbal medicine used worldwide for managing GI tract disturbances. However, its role in gastric cancer is sparingly known. This study ensures the standardization of gastric cancer by the induction of N-nitroso N-methyl Urea (MNU) and to determine the role of the aqueous extract of ginger (AGE) in MNU-induced gastric cancer in albino Wistar rats. Accordingly, the anticancer potential of AGE and its possible mode of action were assessed on rats exposed to MNU, by various biochemical and molecular assays. As evidenced by the extent of lipid peroxidation, gastrin levels and histopathological sections in MNU-induced cancerous lesions at 8 wk which was stabilized at 16 wk confirming the induction of gastric carcinoma by the chemical carcinogen. Further, results revealed that AGE alleviated the oxidative stress as evidenced by the stomach antioxidant enzymes (SOD, catalase, GPx, and GR), markers of oxidative stress (TRx, GRx) and Gastrin, a specific marker for gastric cancer and a decreased level of pro-inflammatory markers (NF-kB, TNF-α, IL-6, PGE2) which was further confirmed by histopathological analysis. AGE is responsible to mitigate oxidative stress and inflammation related to gastric cancer and could be used as a potential dietary intervention in gastric cancer therapy.
Objective: lymph node metastases occur in 90% of cases in patients with gastric adenocarcinoma, and survival is less than 15% in the late stages of disease. Therefore, the purpose of the study was to screening markers for predicting the likelihood of metastases developing in gastric adenocarcinoma. Materials and methods: for the study 47 patients with diagnosis of gastric adenocarcinoma were used (with the localized tumor form T3-4N0M0 (n = 18) and the locally distributed form T3-4N1-3M0 (n = 29)). The copy number variation of 16 genes (APC, AURKA, CCND1, C-MYC, GKN1, HER2, IRX1, MDM2, MET, NFKB, OCT1, P53, PIK3CA, POU5F1B, S6K2 and HV2) was determined by RT-PCR. Immunohistochemical studies were performed on sections of paraffin blocks using monoclonal antibodies to p53, Bcl-2, Ki-67 and Her2. Results: the loss of duplicity of HV2 locus was most frequent CNV in samples of stomach adenocarcinoma, but CNV significant change in the T3-4N0M0 group relative to T3-4N1-3M0 was not detected. A significant increase of CNV in the T3-4N1-3M0 group relative to T3-4N0M0 was found in 2 of 16 loci - NFKB and HER2 in 1.75 and 1.90 times (p<0.0005), respectively. Th e value of p53 expression in the tumor cells nuclei in the group without metastases was lower by 52% compared to the expression in the metastatic group (p<0.05). Low proliferative activity prevailed in the group without metastases in contrast to the group with metastases, in which Her2 overexpression was also observed in 43% of cases in contrast to 20% of cases in the group without metastases. Conclusions: it was showed that the relative copies number of NFKB1 and HER2 in combination with the immunohistochemical parameters of Ki67 and HER2 in gastric tumors can be a prognostic marker of metastasis, and algorithm for predicting metastases development in gastric adenocarcinoma is formed on basis of this.
Background and aims:
Natural honey has been used as a medicine since ancient times. Honey is widely known for its antibacterial properties against H. pylori; however, the mechanisms of its antibacterial activity are not fully known. The present study was performed to examine the molecular mechanisms by which natural honey can inhibit H. pylori infection in gastric epithelial cells.
Methods:
Electrophoretic mobility shift assay was used to measure NF-κB- and AP-1-DNA binding activity. Western blotting was used to detect IκB-α and COX-2 expression.
Results:
H. pylori induced NF-κB and AP-1 DNA-binding activity in gastric epithelial cells. Manuka honey inhibited H. pylori-induced NF-κB and AP-1 in a time- and dose-dependent manner. Maximum inhibition of H. pylori-induced NF-κB and AP-1 by manuka honey was observed at concentrations of 20% at 1-2 h. Pre-treatment of AGS cells with other commercial natural honeys also inhibited H. pylori-induced NF-κB and AP-1 DNA-binding activity. Honey prevented H. pylori-induced degradation of IκB-α protein and downregulated COX-2 protein levels.
Conclusions:
Our findings suggest that natural honey exerts its inhibitory effects against H. pylori by inhibiting NF-κB and AP-1 activation and downregulation of COX-2 expression. These results provide new mechanistic insights into honey effects in the suppression of H. pylori infection.
OBJECTIVE: Methotrexate (MTX)-associated neurotoxicity is an important clinical problem in cancer patients, but the mechanisms of
MTX-induced neurotoxicity are not yet known exactly. The aims of this study were (1) to investigate the possible role of malondialdehyde
(MDA), superoxide dismutase (SOD) enzyme, glutathione peroxidase (GSH-Px) and catalase (CAT) in the pathogenesis of
MTX-induced neurotoxicity and (2) to determine whether there is a putative protective effect of caffeic acid phenethyl ester (CAPE)
on MTX-induced neurotoxicity in the spinal cord, brainstem and sciatic nerve of rats.
METHODS: A total of 19 adult Wistar male rats were divided into three experimental groups. Group I, control group;
Group II, MTX-treated group; and Group III, MTX + CAPE-treated group. MTX was administered to the MTX and MTX + CAPE groups
intraperitoneally (IP) with a single dose of 20 mg/kg on the second day of the experiment. CAPE was administered to the MTX
+ CAPE group IP with a dose of 10 μmol/kg for 7 days.
RESULTS: In the sciatic nerve and spinal cord tissue, CAT and GSH-Px activities were increased in the MTX group in comparison with
the control group. CAPE treatment with MTX significantly decreased CAT and GSH-Px activities in the neuronal tissues of rats in comparison
with the MTX group. In the spinal cord and brainstem tissues, SOD activity in the MTX group was decreased in comparison
with the control group, but in the sciatic nerve, there was no significant difference. In the spinal cord and brainstem of rats, SOD activity
was increased in the CAPE + MTX group when compared with the MTX group. The level of MDA was higher in the MTX group
than in the control group. CAPE administration with MTX injection caused a significant decrease in MDA level when compared
with the MTX group.
CONCLUSION: These results reveal that MTX increases oxidative stress in the sciatic nerve, spinal cord and brainstem of rats and that
CAPE has a preventive effect on the oxidative stress via its antioxidant capacity
Objective: To investigate the expression of nuclear factor kappa-light-chain-enhancer of activated B cell (NF-κB) and Helicobacter pylori (HP) infection status with the use of gastric cancer samples and statistical analysis of HP infection status and clinical data, as well as to provide clinical diagnosis and determine gastric cancer treatment procedures. Methods: T total of 70 paraffin-embedded samples of gastric cancer and 56 normal tissues from surgically resected tissues were randomly selected. NF-κB protein expression in the samples was detected by immunohistochemical staining. A total of 8 living specimens (gastric carcinoma and corresponding normal tissues, 4 pairs of HP+, and 4 pairs of HP) were also selected. The mRNA and of NF-κB protein expression in the samples was detected by quality polymerase chain reaction and western blot. Results: The positive rates of NF-κB protein expression in HP+ gastric carcinoma tissues and adjacent nonneoplastic tissues were 82.50% and 26.32%, respectively. A significant difference in the expression levels was indicated (χ2=24.87, P<0.05, respectively). The positive rates of NF-κB protein expression in HP+ and HP- gastric carcinoma tissues were 82.50% and 53.33% respectively. These results indicated a significant difference in the expression levels (χ2=6.94, P<0.05, respectively). The NF-κB protein expression in gastric carcinoma tissues was 70.00% (49/70). A significant difference in the expression levels was indicated (P<0.05). It correlated with tumor metastasis and invasion but not with histopathologic grading and tumor size. Compared with the adjacent nonneoplastic tissues, HP+ gastric cancer tissues exhibited higher mRNA and NF-κB protein expression in living specimens. However, no significant difference was indicated between HP+ gastric carcinoma tissues with the adjacent nonneoplastic tissues. Conclusion: NF-κB protein expression was upregulated in the HP+ gastric carcinoma sample but not in the HP gastric cancer, HP+, or HP adjacent nonneoplastic tissues. High NF-κB protein expression correlates with gastric cancer development.
Purpose: Fibromyalgia is a chronic pain disorder and its etiology is not well understood. Methods: The relationships among handedness score, Beck depression points and serum S100B levels in fibromyalgia patients were determined and compared between fibromyalgia patients and healthy controls. Results: The rate of left handedness was 10.81%. There were no statistically significant correlations among handedness score, Beck depression point and serum S100B level. Serum S100B levels and depression points were increased in patients with fibromyalgia compared with controls. The high S100B levels in patients with fibromyalgia may have diagnostic and prognostic value in monitoring of fibromyalgia syndromes. Conclusion: These results show that fibromyalgia syndrome is not a cerebral lateralization abnormality.
Purpose: Higher LF/HF ratio and lower heart rate variability have been reported among shift compared with non-shift workers. We aimed to investigate the possible harmful effects of sleep deprivation on cardiac rhythm, specifically heart rate variability (HRV), in work shift physicians. Patients and Methods: Eighty seven healthy male physicians participated in this study. The present study was done in Ankara, Turkey, from January to September 2014. Work shift (sleep deprivation) group (n=45) remained awake for 26 h. Non-work shift group slept in their homes. ECG (HRV) was applied at 9 am for both shift and non-shift groups. Results: Almost all HRV parameters, except LF/HF, decreased in the work shift group compared with the non-shift group. Conclusions: Sleep deprivation due to work shift may cause sympathovagal imbalances by affecting the biological rhythm.
Purpose: The aim of this study was to determine vitamin D levels in children and adolescents aged between 2 months and 16 years, and to investigate risk factors leading to deficient or insufficient vitamin D levels. Patients and Methods: One thousand and ten patients from the pediatric clinic of Turgut Ozal University Hospital in Turkey between January 2010 and May 2014 were enrolled in the study. Patients were divided into three groups: 25(OH) vitamin D deficiency, insufficiency and sufficiency according to their 25(OH) vitamin D levels (≤15 ng/mL, 15-20 ng/mL and ≥20 ng/mL, respectively). 25(OH) Vitamin D levels were measured by HPLC. Results: Approximately a quarter of the children (24.3%) had 25(OH) vitamin D deficiency and 16.5% had 25(OH) vitamin D insufficiency; leaving only (59.2%) of the subjects with sufficient 25(OH) vitamin D levels. Risk factors for 25(OH) vitamin D deficiency were identified by logistic regression analysis using variables of age, gender and season. Logistic regression analysis revealed that winter, spring and autumn seasons and female gender are independent risk factors for 25(OH) vitamin D deficiency/insufficiency (odds ratios: 5.85, 3.93, 1.62 and 1.44, respectively). 25(OH) vitamin D levels decreased as subjects age giving increased odds ratios for subjects aged 13-48 months old (OR: 3.69), 49-108 months old (OR: 4.48) and 109-192 months old (OR: 6.70) compared with 2-12 months old. Conclusion: 25(OH) vitamin D deficiency is a common health problem in childhood. Implementing of 25(OH) vitamin D supplementation should be considered after infancy.
Purpose: The effectiveness of active dry cupping of the upper shoulder and neck to alleviate pain caused by office workers will be investigated. Methods: This randomized, parallel-group trial compared the effectiveness and safety of dry moving cupping therapy for office workers whose neck pain had persisted at least 5 weeks. Those randomized to cupping received up to 10 dry moving cupping therapy sessions over a 5 week period. The study was completed with 40 healthy women subjects who worked at one univeristy in Turkey, with 20 as control and 20 as the study group. Results: Participants mean score of pain on neck was 5.55 (SD:0.57) for pre-test and 2.7 (SD:0.27) for post-test. The decrease of score of pain between pre- and post-test was statistically significant (t=10.14, p=0.002). In the control group there was no significant change in pain score in the statistical significant (t=0.326, p=0.748). Conclusions: Cupping therapy is a non-invasive and harmless therapeutic application and it can be confidently used to reduce the upper shoulder and neck pain in office. It should be considered for all musculoskeletal pain conditions as a complementto medical treatment.
Purpose: Cancer stem cells are resistant to chemotherapy and radiotherapy and are implicated in tumor relapse and high patient mortality. Substances impairing cancer stem cell activity could be very useful as novel cancer therapeutics. Caffeic acid phenethyl ester (CAPE), a component of propolis, whose antitumor activity has been confirmed in several tumor types in vitro, has recently been shown to inhibit the growth of some cancer stem cell types. Methods: An ALDH1-positive fraction was isolated from UT-SCC-74A cells as cancer stem cells by magnetic-activated cell sorting. The isolated cells were characterized by sphere formation assay and biomarkers of cancer stem cells were analyzed by quantitative RT-PCR. To demonstrate the effect of CAPE on head and neck squamous cancer stem cells, XTT cell viability assays were performed. Results: Increasing concentrations of CAPE decrease the viability of head and neck squamous cancer stem cells. IC50 value was calculated from XTT results as 50.29 μg/ml. Conclusion: CAPE has a growth inhibitory effect on head and neck squamous cancer stem cells in vitro.
Purpose: Reflexology activate all body systems physically, mentally and emotionally by relieving pain and relaxing the body, in a manner similar to acupuncture and cupping therapies. The aim of this study was to examine the effect of reflexological therapy on heart rate variability (HRV) parameters in a healthy population. Methods: Participants were twenty-six healthy subjects (8 women, 18 men, median age=32.77 years, SD=8.04). All reflexology procedures were applied by a reflexology practitioner. The reflexology practitioner used thumbs and fingers to apply appropriate pressure to reflexology points, especially the heart point in both feet. The recording ECG was applied 1 hour before and 1 hour after reflexological therapy. Subjects rested for 10 minutes without recording ECG in order to stabilize autonomic parameters. The digital signals were then transferred to a laptop and analyzed using LabChart® software (MLS310/7 HRV Module). Results: Almost all HRV parameters increased and heart (pulse) rate and LF/HF ratio decreased after reflexological therapy compared with before reflexological therapy in healthy persons. Conclusions: These results indicate for the first time in humans that reflexology might induce a state of balance between sympathetic and parasympathetic systems and might be helpful to prevent possible cardiac arrhythmias. Therefore, reflexology may be accepted as a complementary therapy method for many cardiac problems, especially tachycardia and other cardiac arrhythmias.
Purpose: Wet cupping (hijamah) therapy relieves the pain and relaxes the body. The research team aimed to examine the efficacy of wet cupping therapy on the sleep quality parameters in a healthy population. Methods: Participants were seventy-five healthy subjects (50 women, 25 men, median age = 29.61 years, SD = 8.92). Wet cupping was applied by a cupping practitioner. The assessment of the sleep quality parameters by Pittsburg Sleep Quality Index Inventory was applied one hour before and one month after wet cupping therapy. Results: All sleep quality parameters were positively affected after wet cupping therapy in healthy subjects. Conclusions: Wet cupping therapy should be considered as a complementary therapy method for people with sleep problems.
Purpose: Autism is a neurodevelopmental disorder characterized by deficits in social interaction.. It is referred to as cerebral lateralization abnormality. The purpose of this study was to investigate if the different unilateral or lateralized therapeutic applications associated with left cerebral lateralization decreased autistic symptoms. Methods: Autistic symptoms were assessed in six autistic children following stimulation and/or evoking techniques specifically for the left cerebral hemisphere. Results: The use of left cerebral hemisphere stimulation and/ or evoking techniques decreased the autistic symptoms in four of the children with autism. Conclusions: These approaches show promise as a technique to restore dominance delay in the left hemisphere in children with autism and can be useful as a complementary treatment to the other modern methods.
Purpose: Alzheimer's disease (AD), the most common form of dementia, is characterized by abnormal protein storage in the brain and primarily causes a progressive loss of memory and all other cognitive functions. According to the World Health Organization (WHO) report, AD is steadily increasing among people 65 years of age and over. Genome wide association studies have shown that various gene polymorphisms are highly associated with the occurrence of AD. Among them, clusterin (CLU) gene polymorphism was identified as one of the highest genetic risks in late-onset AD. Our aim was to investigate the relation of CLU rs11136000 and AD and its potential use as a biomarker for AD susceptibility in the Turkish population. Methods: 50 samples obtained from AD patients and 55 samples obtained from a control group were used for the presented study. Genomic DNA was isolated from peripheral blood by SDS/proteinase K treatment followed by phenol-chloroform extraction and ethanol precipitation. Presence of the CLU rs11136000 polymorphism was investigated by PCR-RFLP and selected samples were confirmed by DNA sequencing. Chi-square test was used for statistical analysis. Results: In the Turkish population, the CLU rs11136000 polymorphism did not show a significant association with AD as compared with the control group (p>0.05). Polymorphic CLU-C allele of AD patients showed an increased association with diabetes (p=0.015) as compared with CLU-T allele of AD patients, whereas in the control group the CLU-C allele did not show a significant association with diabetes (p=0.332). Conclusion: Individuals with diabetes and polymorphic CLUC allele may have a higher susceptibility to develop AD later in life.
Turgut Ozal University Scientific Research Committee (TOBAT) was established in Turgut Ozal University Faculty of Medicine in 2009 to encourage young medical students and scientists to carry out novel scientific research in addition to their medical education in order to 1) establish a platform of informing the latest advancements in science, 2) present this work to colleagues and 3) meet and interact with their peers within the international medical and scientific community. Our committee annually organizes Turkey's most highly-qualified medical student congress with the highest number of presenters and attendants, the International Medical Student Congress (IMSC). Over 1,500 medical students and experts attend our congress to present, learn and discuss new research in medicine. Medical students from all over Turkey present the results from their scientific work of the previous year. Because of the international nature of this congress, experts, scientists and attendants from other countries enrich the content and atmosphere of the congress. We also invite successful students who have trained in their own countries and who are interested in science and medicine. Students from abroad present their splendid work and also strengthen the global student network. The congress is covered by the media, both print and television, and has a positive impact on public opinion. The conference organization, design and coordination and the configuration of the scientific program are completed by Turgut Ozal University's medical students with the assistance of their supervisors; which is a success in itself. Research and scientific work performed by students compose the main portion of the congress. The latest congress included 161 oral and 74 posters presentations. The topics covered by these presentations often show the promise of playing an important role in the future of medical research. Over the past years, topics have included the following: "CRISPR/Cas9 system": its utilities and its possible applications, especially for tuberculosis infection; the use of synthetic biology to re-program heart coronary arteries for "the rapid treatment of myocardial infarction"; tissue engineering and its novel approaches; and, a state-of-art method for "the colon cancer therapy by using synthetic gut flora". This year, for the first time, we have included a very important opportunity for the congress attendees: 20 studies of the participating scientific presentations have been selected to be published in this supplementary of the Journal of Clinical and Investigative Medicine, a journal cited in SCI and PubMed database. We believe that this opportunity has encouraged the young scientists to improve their research skills, to carry out better and more novel studies and to collaborate for effectively with each other. The selection was difficult because of the high quality of the scientific research. In this supplementary, you will find several well-documented studies on different topics including the genetic roots of Alzheimer's disease related to the clusterin gene, novel approaches for cancer stem cell, a novel reporter protein to be used in lab as an alternative to GFP, the effects of traditional moving dry cupping therapy on sleep quality and shoulder-neck pain and the inhibition of gram negative E.coli by LALF-secreting engineered gram positive B. subtilis. By seeing the quality and breadth of these topics, the contributions and potential impact of our congress to the scientific community can be better understood. We wish to express our sincere appreciation to the editors of the Journal of Clinical and Investigative Medicine for their editing and their support in publishing this supplementary material. We would like also to take this opportunity to thank the Rector of Turgut Ozal University, the Dean of Faculty of Medicine, our instructors, mentors, seniors, advisors and technical staff for their kind advice and assistance in organizing this huge undertaking. Kind regards, Mustafa Semih Elitok on behalf of Turgut Ozal University Scientific Research Committee (TOBAT).
Purpose: The relationship between the number of cytosine-adenine base pair repetition [(CA)n] in the promoter zone of ADAMTS9 gene, which may affect the collagen structure, and the developmental hip dysplasia was investigated. Methods: Using DNA isolating from 26 patients diagnosed with developmental hip dysplasia (DDH) and 29 patients without DDH, the (CA)n in the promoter zone of ADAMTS9 was calculated. The distributions of groups were measured by Shapiro-Wilk test, and the average results of the groups in the state of their normal distributions were compared using parametric Student t test. Results: While the (CA)n values varied between 15 and 30, the group average was found to be 17.9. While the (CA)n values of the patient group varied between 11 and 20, its average was computed to be 17.3. The averages of groups were not statistically different (p = 0.960). Conclusion: According to the results of our study, ADAMTS9 gene promoter (CA)n in the individuals with developmental hip dysplasia was not found to be different from that of healthy individuals. Nevertheless, when strong impact of ADAMTS9 gene on the ligament tissue is considered, it is necessary to evaluate ADAMTS9 gene role with different aspects.
Objective To investigate the role of tumor necrosis factor-α (TNF-α) in epithelial-mesenchymal transition (EMT) of hepatocellular carcinoma (HCC) cells and the related mechanism. Methods HCC cell lines Hep3B and SMMC-7721 were treated with TNF-α for 24 h; then the cell morphological changes were observed by microscope and the expressions of the epithelial markers (E- cadherin and β-catenin), mesenchymal markers (Vimentin and N-cadherin), and EMT associated transcriptional factor Twist were examined by RT-PCR and Western blotting analysis. The invasion and metastasis ability was evaluated by Transwell and wound healing assay. Luciferase reporter assay and immunofluorescence were used to determine NF-κB transcriptional activity; Western blotting analysis was used to examine the expression levels of IκBα and p-IκBα protien. Cells was also incubated with TNF-α and NF-κB inhibitor (BAY11-7082) together, and then the phenotypeof EMT was detected by microscope, RT-PCR and Western blotting analysis. Results Hep3B and SMMC-7721 cells had EMT phenotype after treated with TNF-α. Wound healing assay showed that the wound healing rate of cells exposed to TNF-α was significantly increased compared with the non-treated group (P<0. 05), and Transwell assay showed that more cells penetrating the membrane after treatment with TNF-α (P < 0. 05). TNF-α effectively promoted IκBα phosphorylation and the subsequent NF-κB nuclear translocation We also found that TNF-α-mediated EMT could be converted by NF-κB inhibitor (BAY11-7082) (P<0. 05). Conclusion TNF-α induces EMT of HCC cells through NF-κB-dependent pathways, and subsequently promotes the invasion and metastasis of HCC.
Interleukin (IL)-1 beta has been reported to be a marker of shorter survival in gastric and colorectal adenocarcinoma. In the present study, we examined the potential role and prognostic value of IL-1 beta in esophageal squamous cell carcinoma (SCC). Human esophageal SCC cell line, CE81T, was selected for cellular and animal experiments, in which biological changes after experimental manipulation of IL-1 beta signaling were explored, including tumor growth, invasion capacity, and the sensitivity to treatment. Moreover, 147 esophageal SCC samples were analyzed using immunohistochemical staining to correlate the expression of IL-1 beta with clinical outcome. Our data revealed that IL-1 beta was significantly overexpressed both at mRNA and protein levels in cancer specimens compared to nonmalignant tissues. When IL-1 beta signaling was blocked, tumor growth, invasion ability, and treatment resistance were attenuated. Activation of NF-kappa B, increase of E2-EPF ubiquitin carrier protein and subsequent epithelial-mesenchymal transition might be the underlying mechanisms of the more aggressive tumor growth in IL-1 beta-positive esophageal cancer. The immunochemistry findings indicate that positivity staining of IL-1 beta correlated significantly with higher clinical stage, lower response rate to concurrent chemoradiotherapy (CCRT), and higher recurrence rate after curative treatment. Moreover, IL-1 beta was a significant predictor of survival in patients undergoing surgical intervention or definite CCRT. In conclusion, IL-1 beta is significantly linked to poor prognosis for patients with esophageal cancer and may be a promising molecular target for therapeutic intervention for esophageal SCC.
The ribosomal protein S27 (metallopanstimulin-1, MPS-1) has been reported to be a multifunctional protein, with increased expression in a number of cancers. We reported previously that MPS-1 was highly expressed in human gastric cancer. Knockdown of MPS-1 led to spontaneous apoptosis and repressed proliferation of human gastric cancer cells in vitro and in vivo. However, how does MPS-1 regulate these processes is unclear. Here we performed microarray and pathway analyses to investigate possible pathways involved in MPS-1 knockdown-induced apoptosis in gastric cancer cells. Our results showed that knockdown of MPS-1 inhibited NF-κB activity by reducing phosphorylation of p65 at Ser536 and IκBα at Ser32, inhibiting NF-κB nuclear translocation, and down-regulating its DNA binding activity. Furthermore, data-mining the Gene-Regulatory-Network revealed that growth arrest DNA damage inducible gene 45β (Gadd45β), a direct NF-κB target gene, played a critical role in MPS-1 knockdown-induced apoptosis. Over-expression of Gadd45β inhibited MPS-1 knockdown-induced apoptosis via inhibition of JNK phosphorylation. Taken together, these data revealed a novel pathway, the MPS-1/NF-κB/Gadd45β signal pathway, played an important role in MPS-1 knockdown-induced apoptosis of gastric cancer cells. This study sheds new light on the role of MPS-1/NF-κB in apoptosis and the possible use of MPS-1 targeting strategy in the treatment of gastric cancer.
Cancer stem cells (CSC) are chemoresistant and implicated in tumor recurrence, metastasis and high patient mortality; thus substances impairing CSC activity, could be invaluable as novel cancer therapeutics. We previously showed that CAPE (caffeic acid phenethyl ester), a component of propolis, a honeybee product, inhibits growth of MDA-MB-231 (MDA-231) cells, mdr gene expression, NF-κB, EGFR, and VEGF. We hypothesized that CAPE also acts by interfering with CSC-mediated effects. We isolated breast CSC (bCSC) from MDA-231 cells, a model of human triple-negative breast cancer, and mouse xenografts. bCSC grow as mammospheres (MMS) and when dissociated into single cells, form MMS again, a sign of self-renewal. bCSC exhibited the characteristic CD44(+)/CD24(-/low) phenotype and generated progenitors in the presence of serum, a CSC trait responsible for regenerating tumor mass. CAPE caused dose-dependent bCSC self-renewal inhibition and progenitor formation. Clonal growth on soft agar was inhibited dose-dependently, but apoptosis was not induced as determined by Annexin-V/PI assay. Instead, bCSC were noted to significantly progress from a quiescent cell cycle state in G0/G1 (82%), S phase (12%) to a cycling state with an increase in S phase (41%) and subsequent decrease in G0/G1 (54%). Treatment of bCSC with CAPE (4.5-days) decreased CD44 levels by 95%, while another cell population containing 10-100-fold lower CD44 content concurrently increased. Results suggest that CAPE causes pronounced changes in bCSC characteristics manifested by inhibition of self renewal, progenitor formation, clonal growth in soft agar, and concurrent significant decrease in CD44 content, all signs of decreased malignancy potential.
Coagulation abnormalities are common findings in acute liver failure (ALF) that underlie its fatal outcome. The present study aimed to investigate the effect of caffeic acid phenethyl ester (CAPE) on the oxidative stress and the disturbed hemostasis in lipopolysaccharide/galactosamine-induced ALF in rats. Fifty male Wistar rats were divided into control, ALF and a CAPE-treated ALF groups. Liver transaminases (serum glutamic oxaloacetic transaminase, serum glutamic pyruvic transaminase), total bilirubin, prothrombin time, activated partial thromboplastin time, thrombin time, fibrinogen level, anti-thrombin III levels, platelet count and antioxidative enzymes; catalase and superoxide dismutase activities in serum and reduced glutathione levels in serum and liver tissue were investigated. The ALF group showed increased serum glutamic oxaloacetic transaminase, serum glutamic pyruvic transaminase and total bilirubin. Coagulation tests showed prolonged prothrombin time, activated partial thromboplastin time, thrombin time, decreased platelet count, fibrinogen and anti-thrombin III levels, and decreased activities of the antioxidative enzymes catalase and superoxide dismutase and decreased tissue and serum glutathione levels. Pretreatment with CAPE for 14 days before and a single dose after the exposure to the lipopolysaccharide/D-galactosamine reversed the abnormal liver functions and blood coagulation tests and increased the antioxidative enzymes activities and glutathione levels. CAPE is a promising compound that can support the liver, counteract oxidative stress and disturbed hemostasis in endotoxic-induced ALF.
Intestinal type gastric cancer is a significant cause of mortality, therefore a better understanding of its molecular basis is required. We assessed if either aneuploidy or activity of the oncogenic transcription factor nuclear factor kappa B (NF-kappaB), increased incrementally during pre-malignant gastric histological progression and also if they correlated with each other in patient samples, as they are both induced by oxygen free radicals. In a prospective study of 54 (aneuploidy) and 59 (NF-kappaB) consecutive patients, aneuploidy was assessed by interphase fluorescent in situ hybridisation (FISH) for chromosome 1. NF-kappaB was assessed by expression of interleukin-8 (IL-8), and in a subset, by immunohistochemistry (IHC) for active p65. Aneuploidy levels increased incrementally across the histological series. 2.76% of cells with normal histology (95% CI, 2.14-3.38%) showed background levels of aneuploidy, this increased to averages of 3.78% (95% CI, 3.21-4.35%), 5.89% (95% CI, 3.72-8.06%) and 7.29% (95% CI, 4.73-9.85%) of cells from patients with gastritis, Helicobacter pylori positive gastritis and atrophy/intestinal metaplasia (IM) respectively. IL-8 expression was only increased in patients with current H. pylori infection. NF-kappaB analysis showed some increased p65 activity in inflamed tissues. IL-8 expression and aneuploidy level were not linked in individual patients. Aneuploidy levels increased incrementally during histological progression; were significantly elevated at very early stages of neoplastic progression and could well be linked to cancer development and used to assess cancer risk. Reactive oxygen species (ROS) induced in early gastric cancer are presumably responsible for the stepwise accumulation of this particular mutation, i.e. aneuploidy. Hence, aneuploidy measured by fluorescent in situ hybridisation (FISH) coupled to brush cytology, would be worthy of consideration as a predictive marker in gastric cancer and could be clinically useful in pre-malignant disease to stratify patients by their cancer risk.
Infection with Helicobacter pylori has been linked with chronic atrophic gastritis, an inflammatory precursor of gastric adenocarcinoma. In a nested case-control study, we explored whether H. pylori infection increases the risk of gastric carcinoma.
From a cohort of 128,992 persons followed since the mid-1960s at a health maintenance organization, 186 patients with gastric carcinoma were selected as case patients and were matched according to age, sex, and race with 186 control subjects without gastric carcinoma. Stored serum samples collected during the 1960s were tested for IgG antibodies to H. pylori by enzyme-linked immunosorbent assay. Data on cigarette use, blood group, ulcer disease, and gastric surgery were obtained from questionnaires administered at enrollment. Tissue sections and pathology reports were reviewed to confirm the histologic results.
The mean time between serum collection and the diagnosis of gastric carcinoma was 14.2 years. Of the 109 patients with confirmed gastric adenocarcinoma (excluding tumors of the gastroesophageal junction), 84 percent had been infected previously with H. pylori, as compared with 61 percent of the matched control subjects (odds ratio, 3.6; 95 percent confidence interval, 1.8 to 7.3). Tumors of the gastroesophageal junction were not linked to H. pylori infection, nor were tumors in the gastric cardia. H. pylori was a particularly strong risk factor for stomach cancer in women (odds ratio, 18) and blacks (odds ratio, 9). A history of gastric surgery was independently associated with the development of cancer (odds ratio, 17; P = 0.03), but a history of peptic ulcer disease was negatively associated with subsequent gastric carcinoma (odds ratio, 0.2; P = 0.02). Neither blood group nor smoking history affected risk.
Infection with H. pylori is associated with an increased risk of gastric adenocarcinoma and may be a cofactor in the pathogenesis of this malignant condition.
Caffeic acid phenethyl ester (CAPE), which is derived from the propolis of bee hives, was shown previously to block tumor promoter- and carcinogen-generated oxidative processes in several assays and to engender differential toxicity to some transformed cells. To study the mechanisms of CAPE-induced differential cytotoxicity, nontumorigenic rat embryo fibroblasts (CREF) and adenovirus (type 5)-transformed CREF cells (Wt3A) were used. As shown by nucleosomal-length DNA degradation, morphological alterations by electron microscopy, in situ labeling of 3'-OH ends, and the appearance of a hypodiploid cell population by bivariant flow cytometry, cell death induced by CAPE in the transformed Wt3A cells was apoptosis. Under the same CAPE treatment conditions, CREF cells transiently growth arrested. Both CREF and Wt3A cells were radioresistant, suggesting deficiencies in the proteins controlling the G1 checkpoint. To explore possible mechanisms of CAPE-induced apoptosis, it was determined whether CAPE-induced toxicity was influenced by the redox state of the cells. Depletion of cellular glutathione (GSH) with buthionine sulfoximine before CAPE treatment caused CREF sensitive to CAPE-induced cell death. GSH levels were also determined in CAPE-treated CREF and Wt3A cells. The GSH level in the CREF cells was unaffected by CAPE, whereas the Wt3A cells showed a significant reduction. When the GSH levels were increased in Wt3A cells by treatment with the reducing agent, N-acetyl-cysteine before CAPE treatment, the Wt3A cells were partially rescued. Furthermore, Bcl2, which protects cells from oxidative stress, had a protective effect against CAPE-induced apoptosis in Wt3A cells. Finally, the sensitivity of Wt3A cells to a known oxidant, hydrogen peroxide (H2O2), was examined. Wt3A cells were killed by H2O2-induced apoptosis, whereas CREF cells remained resistant. When Wt3A cells were treated with catalase, a cellular enzyme that inactivates H2O2, CAPE-induced apoptosis in Wt3A cells was reduced, further proving that Wt3A cells were more sensitive than CREF cells to oxidative stress. These results suggest that CAPE can modulate the redox state of cells. Sensitivity of cells to CAPE-induced cell death may be determined by the loss of normal redox state regulation in transformed cells.
Caffeic acid phenethyl ester (CAPE), an active component of propolis from honeybee hives, is known to have antimitogenic, anticarcinogenic, antiinflammatory, and immunomodulatory properties. The molecular basis for these diverse properties is not known. Since the role of the nuclear factor NF-kappa B in these responses has been documented, we examined the effect of CAPE on this transcription factor. Our results show that the activation of NF-kappa B by tumor necrosis factor (TNF) is completely blocked by CAPE in a dose- and time-dependent manner. Besides TNF, CAPE also inhibited NF-kappa B activation induced by other inflammatory agents including phorbol ester, ceramide, hydrogen peroxide, and okadaic acid. Since the reducing agents reversed the inhibitory effect of CAPE, it suggests the role of critical sulfhydryl groups in NF-kappa B activation. CAPE prevented the translocation of the p65 subunit of NF-kappa B to the nucleus and had no significant effect on TNF-induced I kappa B alpha degradation, but did delay I kappa B alpha resynthesis. The effect of CAPE on inhibition of NF-kappa B binding to the DNA was specific, in as much as binding of other transcription factors including AP-1, Oct-1, and TFIID to their DNA were not affected. When various synthetic structural analogues of CAPE were examined, it was found that a bicyclic, rotationally constrained, 5,6-dihydroxy form was superactive, whereas 6,7-dihydroxy variant was least active. Thus, overall our results demonstrate that CAPE is a potent and a specific inhibitor of NF-kappa B activation and this may provide the molecular basis for its multiple immunomodulatory and antiinflammatory activities.
We investigated the mechanisms by which caffeic acid phenethyl ester (CAPE), a phenolic antioxidant, inhibited the stimulation of prostaglandin (PG) synthesis in cultured human oral epithelial cells and in an animal model of acute inflammation. Treatment of cells with CAPE (2.5 microg/ml) suppressed phorbol ester (12-O-tetradecanoylphorbol-13-acetate; TPA) and calcium ionophore (A23187)-mediated induction of PGE2 synthesis. This relatively low concentration of CAPE did not affect amounts of cyclooxygenase (COX) enzymes. CAPE nonselectively inhibited the activities of baculovirus-expressed hCOX-1 and hCOX-2 enzymes. TPA- and A23187-stimulated release of arachidonic acid from membrane phospholipids was also suppressed by CAPE (4-8 microg/ml). Higher concentrations of CAPE (10-20 microg/ml) suppressed the induction of COX-2 mRNA and protein mediated by TPA. Transient transfections using human COX-2 promoter deletion constructs were performed; the effects of TPA and CAPE were localized to a 124-bp region of the COX-2 promoter. In the rat carrageenan air pouch model of inflammation, CAPE (10-100 mg/kg) caused dose-dependent suppression of PG synthesis. Amounts of COX-2 in the pouch were markedly suppressed by 100 mg/kg CAPE but were unaffected by indomethacin. These data are important for understanding the anticancer and anti-inflammatory properties of CAPE.
The growth and spread of tumour cells depends on adequate vasculature. We have previously reported that the expression of interleukin-8 (IL-8) directly correlates with the vascularity of human gastric carcinomas. To provide evidence for a causal role of IL-8 in angiogenesis and tumorigenicity of human gastric cancer, we used the lipofectin method to stably transfect the human TMK-1 gastric carcinoma cells (low endogenous IL-8) with an IL-8 expression vector or control vector. Transfection with IL-8 did not affect the proliferation of cultured cells, yet the culture supernatants of the transfected (but not control) cells stimulated proliferation of human umbilical vein endothelial cells. The IL-8-transfected and control cells were injected into the gastric wall of nude mice. IL-8-transfected cells produced rapidly growing, highly vascular neoplasms as compared to control cells. These results provide direct evidence for the role of IL-8 in the angiogenesis and tumorigenicity of human gastric carcinomas.
Recent data have expanded the concept that inflammation is a critical component of tumour progression. Many cancers arise from sites of infection, chronic irritation and inflammation. It is now becoming clear that the tumour microenvironment, which is largely orchestrated by inflammatory cells, is an indispensable participant in the neoplastic process, fostering proliferation, survival and migration. In addition, tumour cells have co-opted some of the signalling molecules of the innate immune system, such as selectins, chemokines and their receptors for invasion, migration and metastasis. These insights are fostering new anti-inflammatory therapeutic approaches to cancer development.
Androgen deprivation therapy causes a paradoxical elevation of matrix metalloproteinases (MMPs) including MMP-9 resulting in aggressive tumor phenotype in many patients with prostate cancer. In this study, we have evaluated a novel antisense phosphorodiamidate Morpholino oligomer (PMO) targeted against MMP-9 in models of angiogenesis and in human prostate xenograft in athymic mice. The treatment of androgen-independent DU145 human prostate cells with a 21-mer MMP-9 antisense PMO caused a dose-dependent inhibition of cell proliferation compared to scrambled or MMP-2 antisense PMO at similar concentrations. This was associated with decreases in MMP-9 expression, gelatinolytic activity and increased stability of the insulin-like growth factor-binding protein (IGFBP-3), a proapoptotic factor and MMP-9 substrate. In vitro invasion assays revealed a 40-60% inhibition of DU145 cell invasion in the presence of 25 microM MMP-9 antisense PMO. A significant decrease in endothelial cell migration and vascularization was observed in the Matrigel plug assay in mice when treated intraperitoneally with 300 microg/day MMP-9 antisense for 21 days. In the highly vascular DU145 tumor xenografts, MMP-9 inhibition caused decreased tumor growth with regression in 50% of the animals. Histological analysis revealed increased apoptosis and fibrous tissue deposits in the MMP-9 antisense-treated tumors compared to the scrambled and saline controls. No apparent toxicity or mortality was associated with the MMP-9 PMO treatment. In summary, the MMP-9 antisense PMO inhibited in vitro prostate cancer cell proliferation, invasion and in vivo angiogenesis. These data establish the feasibility of developing a site-directed, nontoxic antisense therapeutic agent for inhibiting local invasion and metastasis.
During their lifetime, cells encounter many life or death situations that challenge their very own existence. Their survival depends on the interplay within a complex yet precisely orchestrated network of proteins. The Rel/NF-kappaB signaling pathway and the transcription factors that it activates have emerged as critical regulators of the apoptotic response. These proteins are best known for the key roles that they play in normal immune and inflammatory responses, but they are also implicated in the control of cell proliferation, differentiation, apoptosis and oncogenesis. In recent years, there has been remarkable progress in understanding the pathways that activate the Rel/NF-kappaB factors and their role in the cell's decision to either fight or surrender to apoptotic challenge. Whereas NF-kappaB is most commonly involved in suppressing apoptosis by transactivating the expression of antiapoptotic genes, it can promote programmed cell death in response to certain death-inducing signals and in certain cell types. This review surveys our current understanding of the role of NF-kappaB in the apoptotic response and focuses on many developments since this topic was last reviewed in Oncogene 4 years ago. These recent findings shed new light on the activity of NF-kappaB as a critical regulator of apoptosis in the immune, hepatic, epidermal and nervous systems, on the mechanisms through which it operates and on its role in tissue development, homoeostasis and cancer.
Caffeic acid phenethyl ester (CAPE) is a biologically active ingredient of honeybee propoplis. The cytotoxicity and radiation sensitization effects of CAPE were evaluated in human lung cancer A549 cells and normal lung fibroblast WI-38 cells. A549 cells treated with 6 microg/ml CAPE showed marked growth inhibition (60%) at 48 hr after treatments. During the same time, the number of viable cells decreased to 46% of the control value. In contrast, WI-38 cells showed 20% growth inhibition with no change in the number of viable cells under the same treatment conditions. At 72 hr after CAPE treatment (6 microg/ml), the percentage of apoptotic cells in A549 cultures increased significantly to 67% and an S/G2 arrest was also detected in the culture. Furthermore, there was a significant decrease in the level of intracellular glutathione and hydrogen peroxide contents within one hr after CAPE treatment, and the expression of cyclin B1 was reduced 6 hr after treatment. The radiation sensitization effect of CAPE on A549 cells was determined from the clonogenic survival curves, and the results showed a small but significant difference in radiation survival between cells treated with or without CAPE. Taken together, our results suggest that the effects of CAPE on differential cytotoxicity, apoptosis, and radiosensitization are associated with glutathione depletion that occurred shortly after treatments.
Our previous studies have clearly shown that the angiogenic enzymes, matrix metalloproteinase (MMP) -2/9, are directly involved in human hepatic tumorigenesis and metastasis and suggest that the MMP-2/9 inhibitors, which have dual inhibitory activities on enzyme activity and transcription, represent the best candidates for achieving tumor regression. Many anti-cancer drugs have strong cellular cytotoxicity and side effects, indicating that strong anti-cancer drugs that have no or minimal cytotoxicity and side effects need to be developed. The specific aim of the present study was to develop powerful anti-cancer drugs with specific tumor regression and anti-metastatic potential having the dual inhibitory activities of specific MMP-2 and -9 enzyme activities and gene transcription at the molecular level. Caffeic acid (CA), a strong and selective MMP-9 activity and transcription inhibitor, was isolated from the plant Euonymus alatus and its derivative, caffeic acid phenethyl ester (CAPE), was synthesized. CA and CAPE selectively inhibited MMP-2 and -9 but not -1, -3, -7, or cathepsin K. Treatment of HepG2 cells with CA (100 microg/mL) and CAPE (5 microg/mL) suppressed phorbol 12-myristate 13-acetate (PMA) -induced MMP-9 expression by inhibiting the function of NF-kappaB, but not AP-1. We confirmed that CA and CAPE suppressed the growth of HepG2 tumor xenografts in nude mice in vivo. The subcutaneous and oral administrations of CA and CAPE significantly reduced the liver metastasis. These results confirm the therapeutic potential of the compounds and suggest that the anti-metastatic and anti-tumor effects of CA and CAPE are mediated through the selective suppression of MMP-9 enzyme activity and transcriptional down-regulation by the dual inhibition of NF-kappaB as well as MMP-9 catalytic activity.
The Helicobacter pylori immunodominant protein, CagA, is associated with severe gastritis and carcinoma. Injection of CagA into gastric epithelial cells by type IV secretion leads to actin-cytoskeletal rearrangements and cell scattering. CagA has been reported to have no role in the induction of transcription factor NF-κB and IL-8, which are crucial determinants for chronic inflammation. Here, we provide several lines of evidence showing that CagA is able to induce IL-8 in a time- and strain-dependent manner. We also show that by exchanging specific cagA genes, high IL-8-inducing H. pylori strains could be converted into low inducing strains and vice versa. Our results suggest that IL-8 release induced by CagA occurs via a Ras→Raf→Mek→Erk→NF-κB signaling pathway in a Shp-2- and c-Met-independent manner. Thus, CagA is a multifunctional protein capable of effecting both actin remodeling and potentiation of chemokine release.
• molecular pathogenesis
• pathogenicity island
• type IV secretion
• virulence
Lung cancer is relatively resistant to radiation treatment and radiation pneumonitis is a major obstacle to increasing the radiation dose. We previously showed that Caffeic acid phenethyl ester (CAPE) induces apoptosis and increases radiosensitivity in lung cancer. To determine whether CAPE, an antioxidant and an inhibitor of NF-kappa B, could be a useful adjuvant agent for lung cancer treatment, we examine the effects of CAPE on irradiated normal lung tissue in this study.
We compared the effects of CAPE on cytotoxicity and intracellular oxidative stress in normal lung fibroblast and a lung cancer cell line. For in vivo analysis, whole thorax radiation (single dose 10 Gy and 20 Gy) was delivered to BALB/c male mice with or without CAPE pretreatment. NF- kappaB activation and the expression levels of acute inflammatory cytokines were evaluated in mice after irradiation.
The in vitro studies showed that CAPE cause no significant cytotoxicity in normal lung as compared to lung cancer cells. This is probably due to the differential effect on the expression of NF-kappa B between normal and malignant lung cells. The results from in vivo study showed that CAPE treatment decreased the expression of inflammatory cytokines including IL-1 alpha and beta, IL-6, TNF-alpha and TGF- beta, after irradiation. Moreover, histological and immunochemical data revealed that CAPE decreased radiation- induced interstitial pneumonitis and TGF-beta expression.
This study suggests that CAPE decreases the cascade of inflammatory responses induced by thoracic irradiation without causing toxicity in normal lung tissue. This provides a rationale for combining CAPE and thoracic radiotherapy for lung cancer treatment in further clinical studies.
The proteasome inhibitor bortezomib (Velcade) was recently approved for the treatment of therapy-refractive multiple myeloma and is under investigation for numerous other types of cancer. A phase I clinical trial in paediatric patients resulted in tolerable toxicity. Since the emergence of chemoresistance represents one of the major drawbacks in cancer therapy, we investigated the influence of bortezomib on multi-drug resistant human neuroblastoma cell lines characterised by P-glycoprotein expression and p53 mutation. Nanomolar concentrations of bortezomib inhibited the cell cycle and induced apoptosis in chemosensitive as well as in chemoresistant cell lines. In vivo growth of chemosensitive and chemoresistant neuroblastoma cell lines was inhibited to a similar extent. In addition, bortezomib inhibited vessel formation in neuroblastoma xenografts. These findings and the favourable toxicity profile of bortezomib in children make it reasonable to further pursue additional development of the drug for the treatment of neuroblastoma and other paediatric solid tumours.
NF-κB is an important transcriptional factor that is involved in multiple cellular responses, such as inflammation and antiapoptosis.
IκB kinase α (IKKα) and IKKβ, which are critical regulators of NF-κB activity, possess various mechanisms for NF-κB activation.
This variability in NF-κB signaling may be associated with distinct inflammatory responses in specific cell types. The gastric
pathogen Helicobacter pylori is known to activate NF-κB. However, the role of IKK in H. pylori infection remains unclear. In this report, we show that H. pylori activates both IKKα and IKKβ in gastric cancer cells and enhances NF-κB signaling in distinct manners. We found that IKKβ
acted as an IκBα kinase during H. pylori infection, whereas IKKα did not. H. pylori induced IKKα nuclear translocation in time-, multiplicity of infection-, and cag pathogenicity island-dependent manners. In contrast, p100 processing, which is a known IKKα activity induced by several cytokines,
was not induced by H. pylori. Both IKKs were responsible for chemokine secretion by infected cells. However, the antiapoptotic effect of H. pylori was merely transduced by IKKβ. Microarray analysis and real-time PCR indicated that both IKKs were involved in the transcriptional
activation of genes associated with inflammation, antiapoptosis, and signal transduction. Our results indicate that H. pylori activates NF-κB via both IKKα and IKKβ using distinct mechanisms. IKKα nuclear translocation induced by H. pylori is indispensable for appropriate inflammatory responses but not for antiapoptosis, which suggests a critical role for IKKα
in gastritis development.
The transcription factor NFkappaB plays a role in cell survival. Apoptosis, programmed cell death, via numerous triggers including death receptor ligand binding is antagonized by NFkappaB activation and potentiated by its inhibition. In the present study, we found that caffeic acid phenethyl ester (CAPE), known to inhibit NFkappaB, induced apoptosis via Fas signal activation in human breast cancer MCF-7 cells. CAPE activated Fas by a Fas ligand (Fas-L)-independent mechanism, induced p53-regulated Bax protein, and activated caspases. CAPE also activated MAPK family proteins p38 and JNK. SB203580, a specific inhibitor of p38 MAPK, partially suppressed CAPE-induced p53 activation, Bax expression, and apoptosis, consistent with a mechanism by which CAPE leads to Bax activation, known to be regulated by p38 and p53. The expression of dominant negative c-Jun, which inhibits the JNK signal, also suppresses CAPE-induced apoptosis, suggesting MAPKs are involved in CAPE-induced apoptosis. The expression of Fas antisense oligomers significantly suppressed the CAPE-induced activations of JNK and p38 and apoptosis as compared with Fas sense oligomers. To ascertain whether these phenomena are attributable to the inhibition of NFkappaB by CAPE, we examined the effect of a truncated form of IkappaBalpha (IkappaBDeltaN) lacking the phosphorylation sites essential for NFkappaB activation. IkappaBDeltaN expression not only inhibited NFkappaB activity but also induced Fas activation, Bax expression, and apoptosis. Our findings demonstrate that NFkappaB inhibition is sufficient to induce apoptosis and that Fas activation plays a role in NFkappaB inhibition-induced apoptosis in MCF-7 cells.
Purpose: We examined the role of interleukin (IL)-1β in activation of nuclear factor κB (NF-κB) and the biological function of activated NF-κB in gastric carcinoma cells.
Experimental Design: Human gastric carcinoma cell line GCTM-1 was used to examine NF-κB activation by immunostaining and electrophoretic mobility shift assay. Matrix metalloproteinase (MMP)-9 expression, which plays an important role in tumor invasion, was assessed by semiquantitative reverse transcription-PCR, Western blotting, and immunostaining. The invasive ability of GCTM-1 cells was measured by Matrigel invasion assay. In vivo expression of IL-1β and MMP-9 and activation of NF-κB in 10 surgically resected gastric carcinoma specimens were examined immunohistochemically.
Results: IL-1β enhanced NF-κB activation, MMP-9 expression, and the invasive ability of GCTM-1. A NF-κB inhibitor, pyrrolidine dithiocarbamate, suppressed both MMP-9 expression and invasiveness of IL-1β-treated GCTM-1 cells. IL-1β did not increase the invasive ability of GCTM-1 cells transfected with MMP-9 antisense oligonucleotide. Concomitant expression of IL-1β and nuclear NF-κB was observed in 3 of 10 gastric carcinoma specimens. Cells producing IL-1β were tumor-infiltrating macrophages in two specimens and gastric carcinoma cells in one specimen.
Conclusions: One of the molecules that may play a role in NF-κB activation in some gastric carcinomas is IL-1β. The present results suggest that IL-1β increases the invasive ability of carcinoma cells through activation of NF-κB and the resulting MMP-9 expression.
NF-kappa B is a ubiquitous transcription factor. Nevertheless, its properties seem to be most extensively exploited in cells of the immune system. Among these properties are NF-kappa B's rapid posttranslational activation in response to many pathogenic signals, its direct participation in cytoplasmic/nuclear signaling, and its potency to activate transcription of a great variety of genes encoding immunologically relevant proteins. In vertebrates, five distinct DNA binding subunits are currently known which might extensively heterodimerize, thereby forming complexes with distinct transcriptional activity, DNA sequence specificity, and cell type- and cell stage-specific distribution. The activity of DNA binding NF-kappa B dimers is tightly controlled by accessory proteins called I kappa B subunits of which there are also five different species currently known in vertebrates. I kappa B proteins inhibit DNA binding and prevent nuclear uptake of NF-kappa B complexes. An exception is the Bcl-3 protein which in addition can function as a transcription activating subunit in th nucleus. Other I kappa B proteins are rather involved in terminating NF-kappa B's activity in the nucleus. The intracellular events that lead to the inactivation of I kappa B, i.e. the activation of NF-kappa B, are complex. They involve phosphorylation and proteolytic reactions and seem to be controlled by the cells' redox status. Interference with the activation or activity of NF-kappa B may be beneficial in suppressing toxic/septic shock, graft-vs-host reactions, acute inflammatory reactions, acute phase response, and radiation damage. The inhibition of NF-kappa B activation by antioxidants and specific protease inhibitors may provide a pharmacological basis for interfering with these acute processes.
The transcription factor NF-kappa B, more than a decade after its discovery, remains an exciting and active area of study. The involvement of NF-kappa B in the expression of numerous cytokines and adhesion molecules has supported its role as an evolutionarily conserved coordinating element in the organism's response to situations of infection, stress, and injury. Recently, significant advances have been made in elucidating the details of the pathways through which signals are transmitted to the NF-kappa B:I kappa B complex in the cytosol. The field now awaits the discovery and characterization of the kinase responsible for the inducible phosphorylation of I kappa B proteins. Another exciting development has been the demonstration that in certain situations NF-kappa B acts as an anti-apoptotic protein; therefore, elucidation of the mechanism by which NF-kappa B protects against cell death is an important goal. Finally, the generation of knockouts of members of the NF-kappa B/I kappa B family has allowed the study of the roles of these proteins in normal development and physiology. In this review, we discuss some of these recent findings and their implications for the study of NF-kappa B.
The role of nuclear factor (NF)-kappa B in the regulation of apoptosis in normal and cancer cells has been extensively studied in recent years. Constitutive NF-kappa B activity in B lymphocytes as well as in Hodgkin's disease and breast cancer cells protects these cells against apoptosis. It has also been reported that NF-kappa B activation by tumor necrosis factor (TNF)-alpha, chemotherapeutic drugs, or ionizing radiations can protect several cell types against apoptosis, suggesting that NF-kappa B could participate in resistance to cancer treatment. These observations were explained by the regulation of antiapoptotic gene expression by NF-kappa B. However, in our experience, inhibition of NF-kappa B activity in several cancer cell lines has a very variable effect on cell mortality, depending on the cell type, the stimulus, and the level of NF-kappa B inhibition. Moreover, in some experimental systems, NF-kappa B activation is required for the onset of apoptosis. Therefore, it is likely that the NF-kappa B antiapoptotic role in response to chemotherapy is cell type- and signal-dependent and that the level of NF-kappa B inhibition is important. These issues will have to be carefully investigated before considering NF-kappa B as a target for genetic or pharmacological anticancer therapies.
Gastric carcinoma cells express potent angiogenic factors including vascular endothelial growth factor (VEGF). We previously reported that interleukin-8 (IL-8) acts as an angiogenic factor for human gastric carcinomas. More recently, we found that IL-8 upregulates matrix metalloproteinase-9 (MMP-9) expression and increases invasive activity of gastric carcinoma cells. The purpose of this study was to determine whether the expression of IL-8 and VEGF correlates with clinicopathological parameters in human gastric carcinomas. IL-8 and VEGF expression levels were measured by an enzyme-linked immunosorbent assay (ELISA) in 56 gastric carcinomas and the surrounding normal mucosa. Macroscopic and histopathological tumour findings, presence of metastasis and prognosis were obtained from the patient records and endoscopic, surgical and pathological reports. IL-8 protein levels were higher in most neoplasms than in the corresponding normal mucosal tissue. In contrast, VEGF expression in the tumours was similar to that in normal mucosa. The IL-8 level in the neoplasms correlated significantly with the depth of invasion, venous invasion and lymphatic invasion. VEGF expression in the tumours correlated well with the depth of invasion and lymph node metastasis. No correlation between IL-8 and VEGF expression in the tumours was observed. The survival rates of patients with tumours displaying high IL-8 and VEGF expression levels were significantly lower (P<0.05) than those of patients with tumours displaying low IL-8 and VEGF expression. The results suggest that IL-8 and VEGF may be independent and important prognostic factors in human gastric carcinomas.
Activation of transcription factor nuclear factor-kappaB (NF-kappaB) has been shown to play a role in cell proliferation, apoptosis, cytokine production, and oncogenesis. The purpose of this study was to determine whether NF-kappaB is constitutively activated in human gastric carcinoma tissues and, if so, to determine any correlation between NF-kappaB activity and clinicopathological features of gastric carcinoma.
NF-kappaB activation was determined by immunohistochemical analysis of formalin-fixed, paraffin-embedded specimens from 64 gastric carcinoma patients. We quantified nuclear staining of RelA as a marker of NF-kappaB activation.
Nuclear translocation of RelA was significantly high in tumor cells in comparison to that in adjacent normal epithelial cells (22.5 +/- 2.4% versus 8.6 +/- 1.5%, P < 0.0001). There was a significant correlation between NF-kappaB activation (nuclear translocation of RelA) and expression of urokinase-type plasminogen activator, an invasion-related factor and target of NF-kappaB in tumor cells (rho = 0.393; P = 0.0013). NF-kappaB activation was correlated with tumor invasion-related clinicopathological features such as lymphatic invasion of tumor cells (P = 0.0126), depth of invasion (P = 0.0539), peritoneal metastases (P = 0.0538), and tumor size (P = 0.0164).
Collectively, the data show that NF-kappaB is constitutively activated in human gastric carcinoma tissues and suggest that NF-kappaB activity is related to tumor progression due to its transcriptional regulation of invasion-related factors such as urokinase-type plasminogen activator.
Matrix metalloproteinases (MMPs) play an important role in inflammation, tumor cell invasion, and metastasis. We found that phorbol-12-myristate-13-acetate (PMA)-stimulated invasion of the hepatocellular carcinoma (HCC) SNU-387 and SNU-398 cells and that PMA induced the secretion of MMP-9 in the cells, but did not induce the secretion of MMP-2. The PMA-induced MMP-9 secretion was abolished by treatment of a pan-protein kinase C (PKC) inhibitor, GF109203X, and an inhibitor of NF-kappaB activation, sulfasalazine, and partly inhibited by treatment of inhibitors of ERK pathway, PD98059 and U0126. In addition, the PMA-stimulated activation of the MMP-9 promoter was completely inhibited by a mutation of the NF-kappaB site within the MMP-9 promoter, but not completely by mutations of two AP-1 sites. Moreover, the MMP-9 induction by HGF and TNF-alpha was also completely inhibited by GF109203X and sulfasalazine, but not by PD98059 and U0126. These data demonstrate that the PKC-dependent NF-kappaB activation is absolute for MMP-9 induction and that the PKC-dependent ERK activation devotes to increase the expression level of MMP-9, in HCC cells.
Infection with Helicobacter pylori (H. pylori) is considered a risk factor for gastric carcinoma. The purpose of this study was to clarify whether H. pylori infection plays a role in progression of gastric carcinoma. We examined the expression of genes encoding angiogenic factors and proteases by human gastric carcinoma cell lines (MKN-1 and TMK-1) co-cultured with or without H. pylori by cDNA microarray analysis. Co-culture with H. pylori increased expression of mRNAs encoding interleukin (IL)-8, vascular endothelial growth factor (VEGF), angiogenin, urokinase-type plasminogen activator (uPA), and metalloproteinase (MMP)-9 by gastric carcinoma cells. Up-regulation of these genes at the mRNA and protein levels was confirmed by Northern blot analysis, semi-quantitative RT-PCR analysis, and ELISA. In vitro angiogenic and collagenase activities of conditioned medium from the gastric carcinoma cells were also stimulated by co-culture with H. pylori. These results indicate that H. pylori infection may regulate angiogenesis and invasion of human gastric carcinoma.
The constitutive activation of nuclear factor kappaB (NFkappaB) helps a variety of tumors to resist apoptosis and desensitizes them to chemotherapy, but the causes are still largely unknown. We have analysed this phenomenon in eight mutant cell lines derived from human 293 cells, selected for NFkappaB-dependent expression of a marker gene, and also in seven tumor-derived cell lines. Conditioned media from all of these cells stimulated the activation of NFkappaB (up to 30-fold) in indicator cells carrying an NFkappaB-responsive reporter. Therefore, secretion of extracellular factors as the cause of constitutive activation seems to be general. The mRNAs encoding several different cytokines and growth factors were greatly overexpressed in the tumor and mutant cells. The pattern of overexpression was distinct in each cell line, indicating that the phenomenon is complex. Two secreted factors whose roles in the constitutive activation of NFkappaB are not well defined were investigated further as pure proteins: transforming growth factor beta2 (TGFbeta2) and fibroblast growth factor 5 (FGF5) were both highly expressed in some mutant clones and tumor cell lines, each activated NFkappaB alone, and the combination was synergistic. Our data indicate that a group of different factors, expressed at abnormally high levels, can contribute singly and synergistically to the constitutive activation of NFkappaB in all of the mutant and tumor cell lines we studied. Since several NFkappaB target genes encode secreted proteins that induce NFkappaB, autocrine loops are likely to be ubiquitously important in the constitutive activation of NFkappaB in cancer. We provide the first evidence of the general, complex, and synergistic activation of NFkappaB in tumor and mutant cell lines through the action of secreted factors and suggest that the same explanation is likely for the constitutive activation of NFkappaB in cancers.
To examine the expression of the transcription factor nuclear factor kappa B (NF-kappaB) in Barrett's epithelium and adenocarcinoma and the impact of NF-kappaB expression on tumor stage and response to neoadjuvant chemotherapy and radiation therapy.
Progression of Barrett's esophagus to adenocarcinoma is associated with a wide range of cellular and molecular abnormalities. Nuclear factor-kappa B (NF-kappaB) regulates several genes involved in inflammatory, immune and apoptotic responses, but its role in esophageal inflammation and tumorigenesis has not been reported.
Mobility shift assay was used to measure NF-kappaB activity in nuclear extracts of fresh-frozen biopsies from tumor and uninvolved tissues (n = 30) and esophageal cell lines OE33, SKGT-4, and OE21. RelA expression was assessed by immunohistochemical staining (n = 97). The NF-kappaB/RelA and IkappaB protein expressions were also examined by Western blotting.
NF-kappaB was not expressed in normal esophageal squamous epithelium, in contrast to increased expression in 40% of patients with Barrett's epithelium. Sixty-one percent of resected tumors (n = 97) displayed NF-kappaB immunoreactivity, and 87.5% of the NF-kappaB-positive tumors were Stage IIb and III compared with only 12.5% of patients with Stage I and IIa disease (P < 0.05). The expression of NF-kappaB inversely correlated with major or complete pathologic responses to neoadjuvant chemotherapy and radiation therapy, with 15/20 (75%) responders in the NF-kappaB-negative group compared with 7/38 (18%) in the NF-kappaB-positive group (P < 0.00001). Moreover, incubation of esophageal cell lines OE33, SKGT-4, and OE21 with deoxycholic acid or low pH induced NF-kappaB expression.
Bile acids and low pH induce NF-kappaB expression in esophageal cell lines. NF-kappaB activation is common in esophageal adenocarcinoma. In patients with Barrett's epithelium and an associated esophageal adenocarcinoma, there is a progressive expression of NF-kappaB through Barrett's tumorigenesis. The absence of NF-kappaB expression in esophageal adenocarcinoma correlates with response to neoadjuvant chemoradiotherapy and may be of value in predicting response to neoadjuvant therapy.
Helicobacter pylori infection leads to significant inflammations in the gastric mucosa, which is closely associated with development of gastric cancer. Heat shock protein 90 (HSP 90) has been revealed to be critical for intracellular signaling that participates in inflammatory response as well as carcinogenesis. In this study, we investigated a regulatory role of HSP 90 in H. pylori-induced IL-8 production. Our results showed that H. pylori stimulated significant phosphorylation of HSP 90 and the phosphorylation was diminished by administration of HSP 90 inhibitor, geldanamycin (GA). Treatment of GA completely inhibited H. pylori-induced IL-8 production due to deactivation of ERK1/2 and NF-kappaB. These results subsequently lead to inactivation of AP-1 and NF-kappaB, which are known to be major transcriptional factors of IL-8. Our data provide important insights that HSP 90 is involved as a crucial regulator in H. pylori-induced IL-8 production and its inhibitor could be potentially used for the inhibition of H. pylori-provoked inflammation.
Because the biological significance of constitutive nuclear factor-kappaB (NF-kappaB) activation in human gastric cancer is unclear, we undertook this study to clarify the regulatory mechanism of NF-kappaB activation and its clinical significance.
Immunohistochemistry for NF-kappaB/RelA was done on 290 human gastric carcinoma specimens placed on tissue array slides. The correlations between NF-kappaB activation and clinicopathologic features, prognosis, Akt activation, tumor suppressor gene expression, or Bcl-2 expression were analyzed. We also did luciferase reporter assay, Western blot analysis, and reverse transcription-PCR using the SNU-216 human gastric cancer cell line transduced with retroviral vectors containing constitutively active Akt or the NF-kappaB repressor mutant of IkappaBalpha.
Nuclear expression of RelA was found in 18% of the gastric carcinomas and was higher in early-stage pathologic tumor-node-metastasis (P = 0.019). A negative correlation was observed between NF-kappaB activation and lymphatic invasion (P = 0.034) and a positive correlation between NF-kappaB activation and overall survival rate of gastric cancer patients (P = 0.0228). In addition, NF-kappaB activation was positively correlated with pAkt (P = 0.047), p16 (P = 0.004), adenomatous polyposis coli (P < 0.001), Smad4 (P = 0.002), and kangai 1 (P < 0.001) expression. An in vitro study showed that NF-kappaB activity in gastric cancer cells is controlled by and controls Akt.
NF-kappaB activation was frequently observed in early-stage gastric carcinoma and was significantly correlated with better prognosis and Akt activation. These findings suggest that NF-kappaB activation is a valuable prognostic variable in gastric carcinoma.
Propolis has been used as a folk medicine and has several proven biological activities. Herbal remedies recommended for cancer therapies in Korea.
Matrix metalloproteinase (MMP)-9-inhibitory activity of propolis has been assessed. CAPE as an acting compound was isolated and molecular structure was determined. Anti-invasion activity of CAPE was assayed using hepatocarcinoma cells.
Propolis ethanol extracts showed a strong inhibitory effect of MMP-9 activity, which is known to be involved in tumor cell invasion and metastasis in a concentration-dependent manner on zymography. Assay guided fractionation led to the isolation of a caffeic acid phenyl ester (CAPE) as the compound responsible for the anti-MMP-9 activity. CAPE was obtained by reversed-phase HPLC, and its structure was elucidated by fast atom bombardment mass spectrometry and tandem mass spectrometry. The purified CAPE inhibited MMP-9 activity with the IC(50) of 1.0-2.0 nmol/l.
CAPE possesses selective antiproliferative activity toward hepatocaricoma cell line Hep3B, but not primary cultured mouse hepatocytes.
The pathogenesis of gastric cancer (GC) includes a sequence of events that begins with Helicobacter pylori-induced chronic superficial gastritis, progressing towards atrophic gastritis, intestinal metaplasia, dysplasia and eventually GC. The association between H. pylori and GC is supported by experimental data showing a capacity of H. pylori to induce GC in animals, and the results of interventional studies showing that H. pylori eradication can lower the risk of GC and prevent development of pre-cancerous lesions in humans and in experimental animals. The "driving force" of gastric carcinogenesis is a chronic gastric inflammation, whose intensity and localisation depending on bacterial, host and environmental factors, determines the risk of GC. The mechanisms by which chronic inflammation lead to epithelial and pre-cancerous lesions include induction of oxidative stress, perturbation of the epithelial cells proliferation/apoptosis ratio, and cytokine secretion. Several molecular alterations associated with gastric carcinogenesis have also been described.
We evaluated the activity and toxic effects of bortezomib in patients with mantle cell lymphoma.
Thirty patients, including 29 eligible patients, were enrolled; 13 had received no prior chemotherapy. The dose of bortezomib was 1.3 mg/m2 given on days 1, 4, 8 and 11 every 21 days. Response was assessed according to the International Workshop Criteria for non-Hodgkin's lymphoma and toxicity graded using the National Cancer Institute Common Toxicity Criteria version 2.0.
There were 13 responding patients (46.4%; 95% confidence interval=27.5% to 66.1%), including one unconfirmed complete remission. The median response duration was 10 months. Response rates were similar in previously untreated (46.2%) and treated (46.7%) patients. Neurological toxicity and myalgia led to treatment discontinuation in 10 patients after two to seven treatment cycles. Five serious adverse events (including two deaths) associated with fluid retention were observed in the first 12 patients. We subsequently excluded patients with baseline effusions, dyspnea or edema; no further events were seen.
Bortezomib is active in treating patients with mantle cell lymphoma. While cumulative neuromuscular toxic effects limited therapy duration and specific issues related to fluid retention require further evaluation, continued study of this drug in combination regimens is warranted.
We examined the role of interleukin (IL)-1beta in activation of nuclear factor kappaB (NF-kappaB) and the biological function of activated NF-kappaB in gastric carcinoma cells.
Human gastric carcinoma cell line GCTM-1 was used to examine NF-kappaB activation by immunostaining and electrophoretic mobility shift assay. Matrix metalloproteinase (MMP)-9 expression, which plays an important role in tumor invasion, was assessed by semiquantitative reverse transcription-PCR, Western blotting, and immunostaining. The invasive ability of GCTM-1 cells was measured by Matrigel invasion assay. In vivo expression of IL-1beta and MMP-9 and activation of NF-kappaB in 10 surgically resected gastric carcinoma specimens were examined immunohistochemically.
IL-1beta enhanced NF-kappaB activation, MMP-9 expression, and the invasive ability of GCTM-1. A NF-kappaB inhibitor, pyrrolidine dithiocarbamate, suppressed both MMP-9 expression and invasiveness of IL-1beta-treated GCTM-1 cells. IL-1beta did not increase the invasive ability of GCTM-1 cells transfected with MMP-9 antisense oligonucleotide. Concomitant expression of IL-1beta and nuclear NF-kappaB was observed in 3 of 10 gastric carcinoma specimens. Cells producing IL-1beta were tumor-infiltrating macrophages in two specimens and gastric carcinoma cells in one specimen.
One of the molecules that may play a role in NF-kappaB activation in some gastric carcinomas is IL-1beta. The present results suggest that IL-1beta increases the invasive ability of carcinoma cells through activation of