An Investigation Into The Molecular Mechanism Of Resveratrol Inhibition Of Renal Cancer

Objective The goal of this study was to investigate the effect of resveratrol on the proliferation, cell cycle, apoptosis and invasion of Renca cells.Methods Renca cells were treated with different concentrations (0,10,20,50, 100 ?M) of resveratrol at three time points (12,24 and 48 h) and cell viability was assessed using an MTT assay. Renca cells were treated with various concentrations of resveratrol for 12h, then Renca cells were stained with propidium iodide and the cell cycle evaluated by flow cytometry. Renca cells were incubated with varying concentrations of resveratrol for 24h, and stained with Annexin V/PI or JC-1 Dye. The result was evaluated by flow cytometry. Additionally, the expression of the apoptotic related protein were evaluated by western blotting and protein from Renca cells treated with resveratrol for 24 h. The effect of resveratrol on the invasion of Renca cells was determined with transwell assay.Results Resveratrol inhibited cell proliferation in a concentration-and time-dependent manner. Resveratrol inhibits cell progression by arresting the cell cycle at the G2/M phase in a concentration dependent manner. Resveratrol induced Renca cell apoposis and decreased the mitochondrial membrane potential of Renca cells in a concentration dependent fashion. Resveratrol reduced the level of anti-apoptosis protein Bcl-2, Survivin and Mcl-1, while promoting the expression level of pro-apoptosis protein Bax. Resveratrol inhibited Renca cell invasion in a concentration dependent fashionConclusion Resveratrol can inhibit proliferation, arrest the cell cycle, pro-apoptosis, decrease mitochondrial membrane potential, and lower the invasion of Renca cellsObjective The goal of this study was to investigate the molecular mechanism of resveratrol on Renca cells.Methods Renca cells were treated with various concentrations (0,10,20,50, 100 ?M) of resveratrol for 24 h. Western blot were used to evaluate the amount and the phosphorylation level of STAT3, JAK2, AKT and ERKl/2 in Renca cell lysates. Renca cells were transfected with mouse STAT3 siRNA and cell lysates were prepared after transfection. The expression level of apoptosis-and cyclin-related proteins in cell lysates were evaluated by western blotting. Renca cells were pre-incubated with IL-6 (10ng/mL) for 2 h, then various concentrations (0,10,20,50,100 ?M) of resveratrol were add to the medium. After 24 h, the amount and the phosphorylation status of STAT3 and JAK2 proteins in cell lysates were evaluated by western blotting. Renca cell were pre-incubated with IL-6 (10ng/mL) for 2h, then various concentration (0,20,100?M) of resveratrolwere add to the medium. After 24h, Renca cell viability and apoptosis were assessed by MTT assay and Annexin V/PI.Results Resveratrol had no effects on the amountor the phosphorylation status of AKT and ERK1/2. Resveratrol had no effect on total STAT3 and JAK2 protein levels in Renca cells, however it dramatically inhibited the phosphorylation level of STAT3 and JAK2 in a concentration dependent manner. The result from western blotting showed that Bcl-2, Survivin and Cyclin D1 were inhibited in the STAT3 knockout Renca cell compared with control group. IL-6 had no effect on the total protein level of JAK2 and STAT3, while inducing JAK2 and STAT3 activationin Renca cells, thus alleviating the inhibitory effect of resveratrol on the phosphorylation level of JAK2 and STAT3. IL-6 promoted proliferation and anti-apoptosis effect of Renca cell, decreased the inhibition effect of low concentration (20 ?M) of resveratrol on Renca cell, but high concentration (100 ?M)of resveratrol negated the effects of IL-6 on Renca cell with regard to proliferation and induction of Renca cell apoptosis.Conclusion Resveratrol inhibits Renca cells through the JAK2/STAT3 pathway, and STAT3 gene plays a key role in proliferation and apoptosis. Resveratrol can negate the effect of EL-6, with regard to proliferation and induction of Renca cell apoptosis.Objective The goal of this study was to investigate the effect and the molecular mechanism of resveratrol on Renca tumor growth and Renca metastasis in vivoMethods Renca cells were subcutaneously injected in BALB/c mice, and after 6 days, the mice were randomly divided into three groups of 7 animals, (labed as A, B, C). Resveratrol was administered orally every day at 0,20 and 100 mg/kg body weight to group A, B, C respectively. Tumor tissues were measured every other day and the tumor growth curve was drawn. The tumors was surgically resected 20 days after Renca cell injection. Western blot were used to evaluate the expression level of STAT3, p-STAT3, Bcl-2, Survivin, Cyclin D1 and VEGF protein in tumor tissues and examine the CD31 level by immunohistochemical analysis.BALB/c mice were injected with Renca cells though the tail vein to establish a metastasis model of renca cancer. After 6 days, the mice were randomly divided into three group of 10 animals, (labed as D, E, F). Resveratrol were administered orally every day at 0,20 and 100 mg/kg body weight to group D, E, F respectively. The Kaplan-Meier survival curves were draw according to the survival status of the D, E and F groups and evaluated for the effect of resveratrol on survival time. Another group of 21 mice were divided into three group of 7 animals, (labed as G, H, I). Resveratrol were administered orally every day at 0,20 and 100 mg/kg body weight to group G, H, I respectively. The Lung tissues were surgically resected at 20 days after Renca cell injection and stained with hematoxylin eosin to be evaluated for tumor metastasis in lung.Results Resveratrol inhibits Renca tumor growth and metastasis. Compared with controls and 20mg/kg resveratrol group, the 100mg/kg group exhibited a significant inhibitory effect on tumor growth. The antitumor activity of resveratrol correlated with a reduction in STAT3 activity and a reduction in Cyclin D1, Bcl-2 and Survivin protein expression level in tumors. Moreover, resveratrol decreased the expression level of CD31 and VEGF in tumor tissue. Resveratrol can prolonged the median survival time of mice and inhibits the Renca cell metastasis area of lung.Conclusion Resveratrol inhibits STAT3 activity and new vessels in Renca tumors, Renca growth, and the invasionof Renca cells in vivo, prolong the survival time of metastasis model.