20(S)-ginsenoside Rg3 Anti-hepatocellular Carcinoma Via Promoting The Expression Of P16 By Demethylation

Objective:1,To observe the effect of different concentrations of 20?S?-ginsenoside Rg3 on the inhibition rate of human hepatoma SMMC-7721 cells at different time points.2,To observe the effect of 20?S?-ginsenoside Rg3 intervention on the methylation level of p16 promoter and the expression of p16^INK4a protein in human hepatoma SMMC-7721 cells.Methods:Human hepatoma SMMC-7721 cells were cultured in vitro and divided into blank control group and 20?S?-ginsenoside Rg3 intervention groups.20?S?-ginsenoside Rg3 intervention groups were treated with different concentrations of 20?S?-ginsenoside Rg3 at 40,80,160,320 and 640?M.The cell inhibition rate was detected by MTT assay at 24h,48h and 72h respectively.Statistical analysis was performed to obtain the optimal intervention drug concentration and intervention time.The effect of 20?S?-Rg3 on cell migration was observed by cell scratch test.Methylation-specific PCR was used to detect the effect of 20?S?-ginsenoside Rg3 on the methylation level of p16 promoter in human hepatoma SMMC-7721 cells.The expression of p16^INK4a protein in human hepatoma SMMC-7721 cells was detected by Western blot.Results:1,At the same time point,different concentrations of 20?S?-ginsenoside Rg3intervention could inhibit the survival of SMMC-7721 cells in a dose-dependent manner.2,At the same concentration,20?S?-ginsenoside Rg3 intervention for 24h,48h and 72h could inhibit the survival of SMMC-7721 cells in a time-dependent manner.3,The IC50 of different concentrations of 20?S?-ginsenoside Rg3 for 24h,48h and 72h were 236.4?M,161.2?M and 132.7?M,respectively.4,The optimal intervention concentration of 20?S?-ginsenoside Rg3 was 160?M,and the optimal intervention time was 48h.5,Compared with the blank control group,20?S?-Rg3 could significantly inhibit the migration and proliferation of SMMC-7721 cells?p<0.05?.6,20?S?-ginsenoside Rg3 intervention could reduce p16 promoter methylation level in SMMC-7721 cells.7,20?S?-ginsenoside Rg3 intervention significantly up-regulated the expression of p16^INK4a protein in SMMC-7721 cells?p<0.05 vs.blank control group?.Conclusion:1,20?S?-ginsenoside Rg3 could significantly inhibit the abilityof migration and proliferation of SMMC-7721 cells and play a role in anti-hepatocarcinoma.2,The mechanism of 20?S?-ginsenoside Rg3 may be through decreasing the methylation level of p16 promoter and thereby upregulating the expression of p16^INK4aprotein.