Antiproliferative Effect Of Resveratrol Research Articles

Proliferative mechanism of benign prostatic hyperplasia by NLRP3 inflammasome through the complement pathway.

The expressions of complement component C5a and NLRP3 inflammasome and the antiproliferative effect of resveratrol in benign prostatic hyperplasia (BPH) model rat were analyzed to clarify the BPH proliferative mechanism. This study used the pathological stromal-dominant BPH model rat by urogenital sinus implantation (UGS). Expression of C5a, NLRP3, Caspase-1, IL-1β, and IL-18 using rat BPH tissues at 2, 3, and 8 weeks (n = 6, respectively) after UGS implantation were analyzed by qRT-PCR, western blotting analysis, and immunohistochemical (IHC) analysis. Serum IL-1β levels in BPH model and sham rats were measured by enzyme-linked immunosorbent assay. Furthermore, resveratrol, as the NLRP3 pathway inhibitor, was administered to BPH model rat to assess the antiproliferative effect on the BPH proliferative process. The proliferative effect on prostate was evaluated by Ki-67 protein expression. The expression levels of C5a, NLRP3, Caspase-1, IL-1β, and IL-18 in qRT-PCR, western blotting, and IHC were significantly upregulated in BPH tissues compared to control prostate tissues and showed increases with time (all p < 0.05). Serum IL-1β levels in BPH model rats had significantly increased compared to sham rats. On IHC, deposition of C5a, NLRP3, Caspase-1, IL-1β, and IL-18 was abundant in stromal areas of BPH. The administration of resveratrol significantly decreased prostate weight and expressions of NLRP3, IL-1β, IL-18, and Ki-67 (all p < 0.05). NLRP3 inflammasome activation by complement C5a produces IL-1β and IL-18 through Caspase-1 during the BPH proliferative process. NLRP3 inflammasome have the possibilities to be a therapeutic target for BPH proliferation by inhibiting the NLRP3 inflammasome pathway.

Antiproliferative effects of resveratrol and indole-3-carbinol in HPV-associated diseases prevention

The burden of persistent papillomavirus infection on public healthcare is not limited to cervical cancer (CC). At present, cervical cancer continues to be one of the most common types of cancer worldwide, although HPV also causes anogenital warts. As there is no HPV treatment available, nowadays researchers focus on the search for new therapeutic platforms, new agents for HPV-associated diseases relapse prevention after surgical treatment. Significant anticancer effects of resveratrol have been demonstrated in a variety of cancers, including breast cancer, skin cancer, lung cancer, pancreatic cancer, and stomach cancer. In CC cells, resveratrol stimulates apoptosis of cancer cells, increases expression of HPV E6 and E7, and the expression of p53 tumor suppressor protein. Protective effects on CC cells migration and invasion is achieved by inhibiting NF-κB transcription and AP-1 mediated MMP9 expression. Resveratrol demonstrates a whole range of anti-inflammatory effects, ranging from the suppression of the pro-inflammatory cytokines IL-1α, IL-1β, IL-6, IL-17 expression and inflammatory mediator prostaglandin E2, to the suppression of the formation of ROS and NO. I3K has a proven antitumor efficacy based on the molecular mechanisms of pathological cell proliferation and tumor transformation in hormone- dependent tissues, including cervical epithelial cells. Resveratrol and I3K have a wide range of studied positive therapeutic effects that can significantly reduce the likelihood of development and progression of HPV-associated lesions.

Evidence That β1-Integrin Is Required for the Anti-Viability and Anti-Proliferative Effect of Resveratrol in CRC Cells.

The β1-integrin receptor is broadly expressed on tumor and other cells in the tumor microenvironment (TME), and is an unfavorable prognostic factor for cancers. Nature-derived resveratrol has preventive and apoptotic effects on tumors, but whether resveratrol can exert its suppressive actions on TME-induced tumorigenesis through β1-integrin on the surface of CRC cells is still unknown. HCT116 or SW480 cells were exposed to inhibitory antibodies against β1-integrin, bacitracin (selective β1-integrin inhibitor), integrin-binding RGD (Arg-Gly-Asp) peptide, and/or resveratrol. We evaluated the anti-tumor actions and signaling impacts of resveratrol in colorectal cancer (CRC)-TME. We found that resveratrol completely altered the β1-integrin distribution pattern and expression on the surface of CRC cells in TME. Moreover, resveratrol down-regulated CRC cell proliferation, colony formation, viability, and up-regulated apoptosis in a concentration-dependent way. These actions of resveratrol were antagonized mainly by inhibitory antibodies against β1-integrin but not β5-integrin, and by an integrin-binding RGD peptide but not by RGE peptide, and by bacitracin in TME. Similarly, resveratrol-blocked TME-induced p65-NF-kB and its promoted gene markers linked to proliferation (cyclin D1), invasion (focal adhesion kinase, FAK), or apoptosis (caspase-3), were largely abrogated by anti-β1-integrin or RGD peptide, suggesting that β1-integrin is a potential transmission pathway for resveratrol/integrin down-stream signaling in CRC cells. The current results highlight, for the first time, the important gateway role of β1-integrins as signal carriers for resveratrol on the surfaces of HCT116 and SW480 cells, and their functional cooperation for the modulatory effects of resveratrol on TME-promoted tumorigenesis.

Resveratrol suppresses gastric cancer cell proliferation and survival through inhibition of PIM-1 kinase activity

The proviral integration site for Moloney murine leukemia virus (PIM) family of serine/threonine-specific kinases consist of three isoforms, that regulate proliferation, apoptosis, metabolism, invasion, and metastasis of cancer cells. Among these, abnormally elevated kinase activity of PIM-1 contributes to the progression of gastric cancer and predicts poor prognosis and a low survival rate in gastric cancer patients. In the present study, we found that resveratrol, one of the representative chemopreventive and anticarcinogenic phytochemicals, directly binds to PIM-1 and thereby inhibits its catalytic activity in human gastric cancer SNU-601 cells. This resulted in suppression of phosphorylation of the proapoptotic Bad, a known substrate of PIM-1. Resveratrol, by inactivating PIM-1, also inhibited anchorage-independent growth and proliferation of SNU-601 cells. To understand the molecular interaction between resveratrol and PIM-1, we conducted docking simulation and found that resveratrol directly binds to the PIM-1 at the ATP-binding pocket. In conclusion, the proapototic and anti-proliferative effects of resveratrol in gastric cancer cells are likely to be mediated through suppression of PIM-1 kinase activity, which may represent a novel mechanism underlying its chemopreventive and anticarcinogenic actions.

Resveratrol suppresses the myofibroblastic phenotype and fibrosis formation in kidneys via proliferation-related signalling pathways.

Renal fibrosis acts as the common pathway leading to the development of end-stage renal disease. Previous studies have shown that resveratrol has anti-fibrotic activity, but its potential molecular mechanisms of action are not well understood. The anti-fibrotic effects of resveratrol were assayed in a rat model of unilateral ureteral obstruction (UUO) in vivo and in fibroblasts and tubular epithelial cells (TECs) stimulated by TGF-β1 in vitro. Gene and protein expression levels were analysed by PCR, Western blotting, and immunohistochemical staining. Resveratrol inhibits the myofibroblastic phenotype and fibrosis formation in UUO kidneys by targeting fibroblast-myofibroblast differentiation (FMD) and epithelial-mesenchymal transition (EMT). The anti-fibrotic effects of resveratrol correlated with decreased proliferation of TECs in the interstitium and tubules, resulting in suppressed activity of the proliferation-related signalling pathways, including that of the MAPK, PI3K/Akt, Wnt/β-catenin, and JAK2/STAT3 pathways. Resveratrol treatment suppressed TGF-β1-induced FMD and the expression of the myofibroblastic phenotype in fibroblasts in vitro by antagonizing the activation of proliferation-related signalling. Similarly, TGF-β1-mediated overactivation of the proliferation-related signalling in TECs induced EMT, and the myofibroblastic phenotype was suppressed by resveratrol. The anti-fibrotic and anti-proliferative effects of resveratrol were associated with the inactivation of Smad2/3 signalling and resulted in a partial reversal of FMD and EMT and the inhibition of the myofibroblastic phenotype. Resveratrol suppresses the myofibroblastic phenotype and fibrosis formation in vivo and in vitro via proliferation-related pathways, making it a potential therapeutic agent for preventing renal fibrosis.

Resveratrol suppresses human cervical carcinoma cell proliferation and elevates apoptosis via the mitochondrial and p53 signaling pathways.

Numerous studies have demonstrated the apoptotic and anti-proliferative effects of resveratrol, a natural polyphenolic phytoalexin, on various cancer cell lines. However, the effects of resveratrol on the regulation of human cervical carcinoma, and the mechanisms underlying these effects, remain to be elucidated. In the present study, the potential mechanisms underlying the effects of resveratrol in HeLa cervical carcinoma cells were investigated. The results revealed that resveratrol inhibited proliferation and induced apoptosis in HeLa human cervical cancer cells in a dose-dependent and time-dependent manner. Resveratrol induced cell shrinkage in HeLa cells and apoptosis accompanied by the activation of caspase-3 and −9. Furthermore, resveratrol upregulated the expression of the pro-apoptotic B-cell lymphoma (Bcl)-2-associated X protein and downregulated the expression of the anti-apoptotic proteins Bcl-2 and Bcl-extra large in HeLa cells. In addition, p53, a protein that is essential for cell survival and cell cycle progression, exhibited elevated expression levels in resveratrol-treated HeLa cells. Therefore, resveratrol may be a promising novel inhibitor of human cervical cancer.

Antiproliferative effect of Resveratrol and Oxaliplatin on colon cancer cells

Current cancer treatments include radiation, chemotherapy and surgery. Treatment of primary tumors with radiotherapy and/or chemotherapy has been observed to increase the rate of metastasis. While these treatments contribute to overall well‐being, they are a double‐edged sword and may also cause unintended and painful side effects. Consumption of a diet rich in phytochemicals is efficacious in inhibiting colon cancer. Phytochemicals are plant‐based compounds that are usually nontoxic and have significant effects on cancer cells. They also have been used in a synergistic manner with chemotherapy drugs. This study assessed different plant‐based compounds for effect on colon cancer cells and resveratrol in combination with the known chemotherapy drug, oxaliplatin, displayed interesting results. Resveratrol is found in the skin of red grapes and red wine and has shown promise as an anti‐tumor agent. Our studies focused on assessment of varying concentrations of resveratrol or oxaliplatin used as single compounds and oxaliplatin in combination with resveratrol with resveratrol held constant at the 50uM (IC50) concentration. Results indicated that resveratrol or oxalipatin treatment alone elicited anti‐proliferative activity. Oxaliplatin exhibited greater antiproliferative activity than resveratrol alone. In the double compound treatment analysis, resveratrol (held constant at 50uM) plus oxaliplatin showed significant inhibition at lower concentration (1.56uM) than that of either single compound treatment alone. At higher concentrations, no significant differences were found between oxaliplatin and double‐compound treated cells. Preliminary results suggested that resveratrol plus the chemotherapeutic agent oxaliplatin may inhibit at lower concentrations due to synergistic activities. Studies have shown that both compounds reduce expression of the inhibitor of apoptosis protein known as survivin. Therefore, future experiments will be focused on expression level of micro RNA‐16 and micro RNA‐34a that modulates the survivin gene expression by post transcriptional regulation.

Abstract 1176: The effect of resveratrol combined with dichloroacetate (DCA) on breast cancer cell lines and MCF-10a noncancerous cells

Abstract Resveratrol, (3,5,49-trihydroxy-trans-stilbene), was first reported to act as a chemo-preventative anticancer compound in mice by Jang et al. in 1997 and has been shown to induce apoptosis and reduce both cell viability and mitotic index in many cancer cell lines. Earlier work in our lab found that it affects glucose utilization by decreasing glucose uptake by MCF-7 cells. The present study examined combinations of resveratrol with DCA, a compound that affects mitochondrial metabolism (reversing the Warburg Effect). A goal of this study was to determine if DCA might sensitize cancer cells to other chemotherapeutic compounds that affect glucose metabolism, such as resveratrol. Glucose uptake and lactate production by treated cells were measured, and Western blots were used to estimate levels of several proteins, including cleaved PARP and p21. Both resveratrol and DCA were found to reduce cell viability of treated breast cancer cell lines. Resveratrol, but not DCA, was found to induce caspase-3,7, a measure of apoptosis. This suggests that resveratrol and DCA may affect viability via different mechanisms. More importantly, a synergistic effect of these compounds on cell viability was observed, where DCA was found to enhance the anti-proliferative effect of resveratrol. Citation Format: Sarah Piepenbrink, Dan Sisler, Russ Feirer. The effect of resveratrol combined with dichloroacetate (DCA) on breast cancer cell lines and MCF-10a noncancerous cells. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1176. doi:10.1158/1538-7445.AM2015-1176

Resveratrol‐Induced Inhibition of Endothelial Cell Growth in Vitro Is Dependent on VACM‐1/CUL5 NEDD8ylation Status

Expression of the VACM‐1/CUL5 gene, a subunit of an E3 ubiquitin ligase complex, inhibits cellular proliferation in endothelial cell lines in vitro through a mechanism involving its post‐translational modification by an ubiquitin‐like protein, NEDD8. Factors that regulate VACM‐1/CUL5 NEDD8ylation have not been investigated. We have shown that Resveratrol, a naturally occurring phytoalexin, enhances the antiproliferative effect of VACM‐1/CUL5. In this study, we investigated the effect of Resveratrol on VACM‐1/CUL5 and NEDD8 expression and localization in Human Umbilical Vein Endothelial Cells (HUVEC) in vitro. To determine if the antiproliferative effect of Resveratrol is dependent on VACM‐1/CUL5 NEDD8ylation, we used NEDD8‐aldehyde (N8a), an inhibitor of de‐NEDD8ylation. Our growth assay results showed that Resveratrol decreased cell growth, while N8a increased cell growth at 24 hours. Importantly, Resveratrol attenuated the proliferative stimuli of N8a (n = 4, p &lt; 0.05). Immunocytochemistry results showed that after four hours of treatment with Resveratrol, total VACM‐1/CUL5 was increased while NEDD8 signal was decreased (n = 5, p &lt; 0.05). N8a treatment decreased both NEDD8 and VACM‐1/CUL5 nuclear localization (n = 3, p &lt; 0.05). Finally, in cells treated with both Resveratrol and N8a, VACM‐1/CUL5 expression was increased whereas N8a was decreased (p &lt; 0.05, n = 3). In conclusion, our results suggest that the antiproliferative effect of Resveratrol may be dependent on VACM‐1/CUL5 NEDD8ylation status. This work was supported by the Hope College Departments of Biology and Chemistry Summer Hope Academic Research Program.

SIRT2, a multi-talented deacetylase

Sirtuin 2 (SIRT2) is an NAD(+) (nicotinamide adenine dinucleotide)-dependent deacetylase. Studies of this protein have often been divergent, highlighting the dependence of pleiotropic effects of SIRT2 on cellular context. The natural polyphenol resveratrol is known to exert opposite actions on neural cells according to their normal or cancerous status. We have recently shown the involvement of SIRT2 in the antiproliferative effects of resveratrol on primary cultures of human glioblastoma stem cells. SIRT2 could become a new therapeutic target.

Abstract 3285: Synergistic anticancer effects of resveratrol and clofarabine in malignant mesothelioma cells.

Abstract Malignant mesothelioma is highly resistant to apoptosis. Hence, new strategies to enhance the apoptotic signal and overcome resistance to therapeutic drugs are necessary to improve the therapeutic efficacy of malignant mesothelioma. Here, we investigated the effects of the combined treatment of various phytochemicals as a chemo-potentiator in combination with the chemotherapeutic agent clofarabine in malignant mesothelioma MSTO-211H cells and normal mesothelial Met-5A cells. Treatment with resveratrol in combination with clofarabine at a subtoxic concentration produced a synergistic antiproliferative effect and induced apoptotic cell death, as evidenced by increased subG1 peak on flow cytometric analysis, chromatin condensation and nuclear fragmentation on DAPI staining, and DNA laddering on agarose gel electrophoresis, in MSTO-211H cells, but not in Met-5A cells. In MSTO-211H cells, endogenous expression of the transcription factors Nrf2 and Sp1 proteins, several Nrf2 and Sp1-regulated gene products (HO-1, c-Met, and cyclin D1), and Akt phosphorylation, were effectively suppressed by the combined treatment of clofarabine and resveratrol. Furthermore, the nuclear accumulations of Nrf2 and Sp1 were inhibited by the combination treatment. In combination with clofarabine, the ability of resveratrol to increase the contents of the phosphorylated p53 and the apoptotic molecules, PARP and cleaved caspase 3, was also evident in a time- and dose-dependent experiment. Taken together, the data provide evidence that the synergistic antiproliferative effect of resveratrol and clofarabine is linked to simultaneous suppression of multi-target molecules, such as Nrf2, Sp1, and Akt activities, and suggest that this combination may have therapeutic value in treatment of malignant mesothelioma. Citation Format: David M. Lee, Yoon-Jin Lee, Jae-Hyuk Im, Seong-Youn Won, Hae-Seon Nam, Sang-Han Lee. Synergistic anticancer effects of resveratrol and clofarabine in malignant mesothelioma cells. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3285. doi:10.1158/1538-7445.AM2013-3285

Inhibition of cell cycle progression and apoptotic activity of resveratrol in human intrahepatic cholangiocarcinoma cell lines

Background: Cholangiocarcinoma (CCA) is a cancer of the bile duct epithelium and is characterized by a poor prognosis and unresponsiveness to conventional treatments. Effective therapeutic agents for CCA are urgently needed. Resveratrol, a phytoalexin found in several fruits, has been reported to inhibit growth of various cancer cell lines. Objective: We investigated the effect of resveratrol on the intrahepatic CCA-derived KKU-100 and KKU-M156 cell lines. Materials and methods: The cell lines used were established from Thai CCA patients (KKU-100, a poorlydifferentiated adenocarcinoma and KKU-M156, a moderately-differentiated adenocarcinoma). The mechanisms of an antiproliferative effect of resveratrol on these cell lines were determined using sulforhodamine B assay, flow cytometry, ethidium bromide/acridine orange staining, DNA fragmentation, and Western blotting analysis. Results: Resveratrol significantly inhibited CCA cell growth in a dose- and time-dependent manner. Resveratrol induced cell cycle arrest at the G0/G1 phase in KKU-100 by decreasing cyclin D1, cyclin E, cyclin-dependent kinase (Cdk)2 and Cdk4 levels and increasing p53, Cdk inhibitors (CDKIs) p21 and p27 levels. By comparison, resveratrol induced cell cycle arrest at the S and G2 phases in KKU-M156 cells by increasing cyclin E, Cdk2, p53, p21 and p27 levels and decreasing cyclin B1 and Cdk1 levels. Subsequently, resveratrol induced apoptosis of both cell lines by increasing the Bax/Bcl-2 ratio and apoptosis-inducing factor, and decreasing survivin and subsequent activation of caspase-9 and -3 and DNA fragmentation. Conclusion: Our study highlighted for the first time that resveratrol had different effects on cell cycle progression and apoptosis mechanisms in different CCA cell types. Resveratrol inhibits growth of the KKU-100 and KKUM156 cell lines by arresting different phases of the cell cycle and inducing a mitochondrial-dependent apoptosis pathway, through both caspase-dependent and -independent means. These results suggest that resveratrol could be developed as a chemotherapeutic agent against CCA. Keywords: Apoptosis, cell cycle arrest, human cholangiocarcinoma cell lines, mitochondrial-dependent signaling pathway, resveratrol

Abstract 1152: Micro RNA-21 serves as an essential target for resveratrol's anti-tumor action against metastatic prostate cancer cells

Abstract Background: Resveratrol, a phytoalexin, has been reported to possess benefits against various cancers. However, the exact mechanism(s) underlying this beneficial response is not clearly understood. MicroRNA-21 (miR-21) is a small non-coding RNA which represses the translation of various tumor-suppressor genes. miR-21 levels are often upregulated in prostate cancer where it is positively associated with increased proliferation, invasion and metastasis and reduced apoptosis. Here, we demonstrate that resveratrol negatively regulates proliferation, invasion and apoptosis by modulating the levels of miR-21 in an androgen-independent, metastatic prostate cancer line (PC-3MM). Methods: The effect of resveratrol on PC-3MM cell proliferation at different time points was measured using MTS assay. Apoptosis and cell cycle analysis were determined by flow cytometry. The effects of resveratrol on invasion and migration were assessed by Boyden chamber and wound healing assays, respectively. miR-21 levels were determined by real-time PCR. Western blotting was used to assess modulation of miR-21 downstream proteins. In addition, siRNA transfections were performed to suppress the expression of specific proteins in order to elucidate the molecular mechanisms underlying the actions of resveratrol. Results: We observed that resveratrol significantly reduced proliferation of PC-3MM cells in a dose-dependent manner. This anti-proliferative effect of resveratrol was due to the induction of cell cycle arrest in G2/M and S phase and increased apoptosis at 25 and 50 μM doses of resveratrol. Resveratrol also inhibited cell invasion and migration in a dose-dependent manner in Boyden chamber and wound healing assays, respectively. These changes were associated with reduced miR-21 expression and increased expression of miR-21-tumor suppressor targets, such as maspin and program cell death 4 (PDCD4). Inhibition of miR-21 by mir21 antisense oligonucleotide also had the same effect on maspin and PDCD4 expression. Furthermore, resveratrol induced inhibition of proliferation, invasion and migration of PC-3MM cells were reversed by overexpressing pre-miR-21 or knockdown of PDCD4 by siRNA. Conclusion: This study demonstrates, for the first time, that resveratrol can inhibit miR-21 expression which could account for its anti-tumor effects, such as inhibition of proliferation, invasion and migration of PC-3MM prostate cancer cells. Inhibition of miR-21 occurs at doses of resveratrol (25 and 50 μM) which are readily achieved in vivo following consumption of resveratrol in red wine and grapes. Based up on these interesting results, we propose that resveratrol could serve as a potential dietary supplement for cancer chemoprevention or as an adjunct to existing chemotherapeutic regimens for treating prostate cancer. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1152. doi:10.1158/1538-7445.AM2011-1152

Antiproliferative effect of resveratrol in pancreatic cancer cells

To investigate resveratrol, one of the food derived polyphenols that might be partially responsible for the beneficial effect on cancer, the in vitro antitumor activity of resveratrol against pancreatic cancer cell lines (PANC-1, BxPC-3 and AsPC-1) was examined, together with the mechanisms involved. The effects of resveratrol on the growth inhibition, apoptosis and cell cycle were assayed. The activity of caspases and the expression of Bcl-2, Bcl-xL, XIAP and Bax protein were detected. The results showed that resveratrol inhibited the proliferation of pancreatic cancer cells in a dose- and time-dependent manner. Resveratrol inhibited the cell growth of PANC-1, BxPC-3 and AsPC-1 cells with IC(50) values of 78.3 ± 9.6 μmol/L, 76.1 ± 7.8 μmol/L and 123.1 ± 6.5 μmol/L at 48 h, respectively. Incubation of pancreatic cancer cells with resveratrol resulted in cell apoptosis and cell cycle arrests. Resveratrol induced activation of caspases. Simultaneously, resveratrol regulated the expression of the antiapoptotic proteins Bcl-2, Bcl-xL and XIAP and the proapoptotic protein Bax. PANC-1 and BxPC-3 cells were more chemosensitive to resveratrol than AsPC-1 cells. In conclusion, resveratrol inhibited the proliferation of pancreatic cancer cells by inducing apoptotic cell death. There was different sensitivity to resveratrol in different pancreatic cancer cell lines.

Abstract 601: Mechanistic Dichotomy Of Resveratrol-mediated Inhibition Of Vascular Smooth Muscle Cell Proliferation And Restenosis

Angioplasty is a widely used therapy for coronary artery stenosis, but its application is limited by restenosis occuring in 30 – 40% of patients within 3– 6 months of surgery. Restenosis is due to intimal hyperplasia resulting from migration to and proliferation of vascular smooth muscle cells (VSMC) within the intima. Recent epidemiological evidence suggests red wine exerts greater cardioprotection in women than men. Our hypothesis was that the anti-proliferative effects of resveratrol are mediated by estrogen receptor stimulation. We thus determined effects of resveratrol in VSMC isolated from female rats, compared to female VSMC treated with siRNA for estrogen receptor- (ER-)α. At low doses (0.5–25 μM) resveratrol inhibited VSMC proliferation through an ER-α dependent increase in inducible nitric oxide synthase (iNOS) activity and inhibited the cells in S phase, but at high doses (50–100 μM) the mechanism was NF-κB dependent, with inhibition of cells in G 0 /G 1 phase. At all doses, resveratrol inhibited IKK activity, an upstream kinase for NF-κB activation, but its effect at low doses was partially reversed by ER-α blockade. We also found a mechanistic link between ER-α receptors, cyclooxygenase-2 (COX-2), a known modulator of VSMC proliferation, and resveratrol. At low doses, resveratrol induced COX-2 expression, consistent with ER-α stimulation, while at high doses it inhibited COX-2 protein levels, likely due to an inhibition of NF-kB. To confirm these findings in an in vivo model for restenosis, B6.129 (n=8 per group) mice were fed with a high fat diet +/− 50 mg/kg resveratrol for 2 weeks. A carotid artery endothelial denudation procedure was conducted and after recovery, the mice were again fed a high fat diet +/−resveratrol for 2 weeks. Resveratrol increased arterial iNOS activity (p&lt;0.05) and total NO production (p&lt;0.05) but inhibited NF-κB activation (p&lt;0.01). Studies in progress utilizing ER-α (−/−) mice are determining the role of ER-α in resveratrol- mediated effects and whether resveratrol prevents neointimal hyperplasia in this model. In conclusion, though resveratrol clearly inhibited NF-κB activation, it appears that it’s predominate therapeutic effect at low pharmacologically relevant concentrations is via ERα modulation.

Resveratrol reduces oxidation and proliferation of human retinal pigment epithelial cells via extracellular signal-regulated kinase inhibition

Epidemiological evidence suggests that moderate wine consumption and antioxidant-rich diets may protect against age-related macular degeneration (AMD), the leading cause of vision loss among the elderly. Development of AMD and other retinal diseases, such as proliferative vitreoretinopathy (PVR), is associated with oxidative stress in the retinal pigment epithelium (RPE), a cell layer responsible for maintaining the health of the retina by providing structural and nutritional support. We hypothesize that resveratrol, a red wine polyphenol, may be responsible, in part, for the health benefits of moderate red wine consumption on retinal disease. To test this hypothesis, the antioxidant and antiproliferative effects of resveratrol were examined in a human RPE cell line (designated ARPE-19). Cell proliferation was determined using the bromodeoxyuridine (BrdU) assay, intracellular oxidation was assessed by dichlorofluorescein fluorescence, and activation of the mitogen-activated protein kinase (MAPK) cascade was measured by immunoblotting. Treatment with 50 and 100 μmol/L resveratrol significantly reduced proliferation of RPE cells by 10% and 25%, respectively ( P < 0.05). This reduction in proliferation was not associated with resveratrol-induced cytotoxicity. Resveratrol (100 μmol/L) inhibited basal and H 2O 2-induced intracellular oxidation and protected RPE cells from H 2O 2-induced cell death. The observed reduction in cell proliferation was associated with inhibition of mitogen activated protein kinase/ERK (MEK) and extracellular signal-regulated kinase (ERK 1/2) activities at concentrations of resveratrol as low as 5 μmol/L. These results suggest that resveratrol can reduce oxidative stress and hyperproliferation of the RPE.

Resveratrol에 의한 A549 인체 폐암세포의 증식억제 및 apoptosis 유발에 관한 연구

Resveratrol은 포도와 같은 식물에서 각종 감염균으로부터 자신의 몸을 보호하기 위하여 생성되는 물질인 phytoalexin의 일종으로 강력한 항산화작용, 암예방 효과 및 항암 작용을 포함한 각종 약리작용을 가진 것으로 보고되어져 오고 있다. 본 연구에서는 resveratrol의 항암작용 기전해석을 위하여 A549 인체폐암세포의 종식에 미치는 resverakol의 영향을 조사하였다. A549 세포의 생존율은 resveratrol의 처리시간 증가에 따라 강력하게 억제되었으며, 이는 암세포의 다양한 형태변형을 동반한 세포주기 C2/M arrest 및 염색질 응축 현상을 동반한 apoptosis 유발에 의한 것임을 알 수 있었다. Resveratrol 처리에 의한 apoptosis 유발은 Bcl-2의 발현변화 없이 Bcl-<TEX>$X_L$</TEX>의 발현 감소에 따른 상대적인 Bax의 발현 증가와 Sp-1, PCNA 및 <TEX>$\beta$</TEX>-catenin 등과 같은 단백질의 분해 현상과 연관성이 있었다 또한 resveratrol에 의한 A549세포 의 증식억제는 Cdk inhibitor p21의 발현 증가에 따른 Cdks 의 kinase 활성 저하 및 COX-2의 선택적 저해에 따른 PGE2 생성 저하와 관련이 있었다. Resveratrol, a phytoalexin found at high levels in grapes and in grape products such as red wine, has been reported to possess a wide range of biological and pharmacological activities including antioxident, anti-inflammatory, anti-mutagenic, and anti-carcinogenic effects. According to recent studies, this compound is an effective inhibitor of cell growth in general, triggers partial arrest of the cell cycle and induce apoptosis. In this study, the anti-proliferative effects of resveratrol in A549 human lung carcinoma cells were investigated. It is shown that resveratrol induced the growth inhibition in a time-dependent manner and morphological changes of A549 cells, which were associated with induction of S phase arrest of the cell cycle and apoptotic cell death. The Bcl-<TEX>$X_L$</TEX>levels were markedly down-regulated in resveratrol treated cells, however, Bax and Bcl-2 were remained unchanged. Resveratrol treatment induced the proteolytic degradation of Sp-l and proliferating cell nuclear antigen protein, and inhibited the expression of <TEX>$\beta$</TEX>-catenin protein. Resveratrol treatment also induced a marked up-regulation of cyclin-dependent kinase (Cdk) inhibitor p21 and inhibited the kinase activities of Cdk2 and Cdk4. In addition, resveratrol treatment inhibited the levels of cyclooxygenase (COX)-2 mRNA and protein, and the release of prostagladin E2 without alteration of COX-1 expression. Taken together, these findings suggest that resveratrol may be a potential chemotherapeutic agent for the control of human lung carcinorma cells.

Modulations of critical cell cycle regulatory events during chemoprevention of ultraviolet B-mediated responses by resveratrol in SKH-1 hairless mouse skin.

Multiple exposures to solar ultraviolet (UV) radiation cause critical damages that may lead to the development of several cutaneous disorders including skin cancer, the most frequently diagnosed malignancy in the USA. Therefore, efforts are needed to: (i) study the mechanism(s) of UV-mediated cutaneous damages, and (ii) design novel approaches for the management of skin cancer. 'Chemoprevention' via plant-based agents may be a useful approach for the management of neoplasia. Here, we evaluated the involvement of cell cycle regulatory molecules during resveratrol-mediated protection from multiple exposures of UVB (180 mJ/cm(2); on alternate days x 7 exposures) radiations in the SKH-1 hairless mouse skin. Resveratrol was topically applied on the skin of SKH-1 hairless mice at a dose of 10 micromol/mouse (in 0.2 ml acetone; 30 min prior to each UVB exposure). Studies were performed at 24 h following the last UVB exposure. Topical application of resveratrol resulted in significant decrease in UVB-induced bi-fold skin thickness, hyperplasia, and infiltration of leukocytes. The data from immunoblot and/or immunohistochemical analyses revealed that multiple exposure to UVB radiations causes significant upregulation in: (i) proliferating cell nuclear antigen (PCNA), a marker of cellular proliferation, and (ii) cyclin-dependent kinase (cdk)-2, -4 and -6, cyclin-D1, and cyclin-D2. Resveratrol treatment resulted in significant downregulation in UV-mediated increases in these critical cell cycle regulatory proteins. An interesting observation of this study was that resveratrol treatment resulted in a further stimulation of UVB-mediated increases in cyclin kinase inhibitor WAF1/p21 and tumor suppressor p53. Further, resveratrol was also found to cause significant decreases in UVB-mediated upregulation of: (i) the mitogen-activated protein kinase kinase, and (ii) the 42 kDa isotype of mitogen-activated protein kinase (MAPK). Thus, our data suggested that the antiproliferative effects of resveratrol might be mediated via modulation in the expression and function of cell cycle regulatory proteins cyclin-D1 and -D2, cdk-2, -4 and -6, and WAF1/p21. Our data further suggest that the modulation of cki-cyclin-cdk network by resveratrol may be associated with inhibition of the MAPK pathway. We suggest that resveratrol may be useful for the prevention of UVB-mediated cutaneous damages including skin cancer.

Resveratrol inhibits cell proliferation and induces apoptosis of human breast carcinoma MCF-7 cells

Resveratrol, which is found in grapes and wine, has been reported to have a variety of important pharmacological effects including anti-inflammatory, anti-platelet, and anti-carcinogenetic properties. In this study, using the human breast cancer cell line MCF-7, we have analyzed a possible mechanism by which resveratrol could interfere with cell cycle control and induce cell death. Resveratrol treatment of MCF-7 cells resulted in a dose-dependent inhibition of the cell growth and the cells accumulated at the S phase transition of the cell cycle at low concentrations, but high concentrations do not induce S phase accumulation. The anti-proliferative effects of resveratrol were associated with a marked inhibition of cyclin D and cyclin-dependent kinase (Cdk) 4 proteins, and induction of p53 and Cdk inhibitor p21WAF1/CIP. Growth suppression by resveratrol was also due to apoptosis, as seen by the appearance of a sub-G1 fraction and chromatin condensation. In addition, the apoptotic process involves activation of caspase-9, a decrease of Bcl-2 as well as Bcl-XL levels, and an increase of Bax levels.

Antiproliferative effect of resveratrol in human prostate carcinoma cells.

Resveratrol, a polyphenolic phytoalexin found in grapes, may have potential for the prevention and treatment of human cancer. We report here that resveratrol inhibits the growth of human prostate carcinoma DU145 cells and provide a molecular explanation of the effect. Resveratrol treatment in DU145 cells resulted in a dose-dependent inhibition of cell growth and induced apoptotic cell death. The antiproliferative effect of resveratrol was associated with the inhibition of D-type cyclins and cyclin-dependent kinase (Cdk) 4 expression, and the induction of tumor suppressor p53 and Cdk inhibitor p21. Moreover, the kinase activities of cyclin E and Cdk2 were inhibited by resveratrol without alteration of their protein levels. Resveratrol treatment also up-regulated the Bax protein and mRNA expression in a dose-dependent manner; however, Bcl-2 and Bcl-xL levels were not significantly affected. These effects were found to correlate with an activation of caspase-3 and caspase-9. Taken together, our study suggests that resveratrol has a strong potential for development as an agent for the prevention of human prostate cancer.