The Effect And Mechanism Of Resveratrol On HepG2Cells

Hepatocellular carcinoma is one of the most common malignancy in the world and its mortality is the second place in China. The treatment of HCC has made great progress in recent years in China,and the rate of early diagnosis,surgical resection,postoperative survival rate and quality of life have been improved greatly,but the current incidence and mortality are still high.The way of treatment of HCC depends on tumor stage and liver function. When the tumor is unresectable and can not tolerate the surgery,the patient can choose a combination of local and systemic treatment of tumor. Non-surgical treatment has limited efficacy. So,to find effective drugs to improve survival, reduce the relapse rate and an important means of improving quality of life is of great significance.Resveratrol is a non-flavonoid ployphenol which has anti-inflammatory,antioxiodation and cardiovascular protecting.Various studies have shown that resveratrol had a certain degree of inhibition and cytotoxicity to a variety of human tumor cells in vitro,such as liver,stomach,breast,colon cancer,et al. Its medicinal properties have become a hot topic,but the mechanism still needs further study. Charpter1The growth inhibition of Resveratrol on hepG2cells.Objiective:To observe the growth inhibiton and cytotoxicity of Resveratrol on hepG2cells.Methods:MTT assay,direct observation of cell morphology and flow cytometry was performed to observe the growth inhibiton and apoptosis of HepG2cells.Results:(1)MTT:The inhibition rates of each positive control groups were greater than0.3with drug susceptibility positive after24h to48h, and the inhibition rates were statistically significant compared with the negative control group.There was inhibition of resveratrol on HepG2cells, and it increased significantly with the increase of drug concentration(p<0.01).Compared with the inhibition rate of HepG2cells after24h, the inhibition rates with the same concentration of resveratrol of HepG2cells after48h and72h increased(p<0.05),the inhibiton rate of resveratrol on HepG2cells after72h increased compared48h, but there was no statistically significant. The IC50of resveratrol were77.02±5.13μg/ml,22.76±1.78μg/ml and18.22±1.34μg/ml measured after24h to48h. The inhibiton of resveratrol on LO2cells was not obvious, and the inhibitory rate between HepG2and LO2cells treated with resveratrol had a significant difference(p<0.01).(2)Direct observation of cell morohology:Deformation,folds,floating,unclear border were found in HepG2cells when the Res concentration increased,and the majority of cells in5FU group became smaller and more unclear.The change of LO2cells was not obvious(3)Flow cytometry:The apotosis rate of HepG2increased significantly with the Res concentration increasing, and compared to the control group,the drug groups was statistically significant(p<0.05).Conclusion:(1) Res concentration-dependently and time-dependently inhibited growth and promoted apotosis of HepG2cells;(2)The effects of resveratrol on proliferation of LO2cells was not obvious. Charpter2:The mechanism of resveratrol on HepG2cellsObjiective:To detect the expression of MnSOD,SIRT3,P53,Bax and Fas in HepG2cells under Res with different concen-tration.Methods:MnSOD enzyme activity of HepG2cells was detected by WST assay.PT-PCR and Western-blot were performed to detect expression of SIRT3,P53,Bax and Fas in HepG2cells under Res with different concentration.Results:WST:The MnSOD enzyme activity was significantly increased treat with Res,and compared to the control group,the drug groups was statistically significant(p<0.01). RT-PCT:The expression of SIRT3,Fas,Bax and p53in HepG2cells increased(p<0.01) after treated by Res after48h. Western-blot:The expression of SIRT3,Fas,Bax and p53in HepG2cells increased(p<0.01) after treated by Res after48h.Conclusion:Res promote the expression of SIRT3in HepG2cells,and thus increase the activity of MnSOD enzyme and the expression of p53which activate Bax and Fas to induce apoptosis. Total conclusion:(1) Res concentration-dependently and time-dependently inhibited growth and promoted apotosis of HepG2cells;(2) Res promote the expression of SIRT3in HepG2cells,and thus increase the activity of MnSOD enzyme and the expression of p53which activate Bax and Fas to induce apoptosis.