The Effects Of Resveratrol On Proliferation And Apoptosis Of Hepatocellular Carcinoma 7721 Cells

Objective:To observe the effects of resveratrolon the proliferation and apoptosis of hepatocellular carcinoma 7721 cells and to probe the action mechanism bases on the mitochondrial apoptotic pathway.Method:Hepatocellular carcinoma 7721 cells were cultured and treated with the different concentrations of resveratrol?6.25 12.5 25 50 100?mol/L?,and the morphological changes of cells were observed under inverted microscope.MTT assay was used to observe the changes of proliferation of 7721 cells at different time?24,48,72 h?.Flow cytometry was used to analyze the cell cycle changes after 48h.Hoechst 33258 staining method was used to observe apoptosis after48h.Annexin V-FITC/PI double staining method was used to analyze the apoptosis rate after 48h.Western blot was used to verify the expression of Bcl-2,Bax,Caspase-3 and Caspase-9 proteins of mitochondrial apoptosis pathway after 48h.Result:1.Different concentrations of resveratrol?6.25 12.5 25 50 100?mol/L?acted on7721 cells and observation under inverted microscope showed that:morphology is irregular,the cell wall is poorly adherent,the cell number is decreased,and the cell morphology is even round.floating with the drug concentration increases.2.The above different concentrations of resveratrol acted on 7721 cells after 24,48 and 72 hours and MTT assay results showed that:resveratrol can inhibit the proliferation of hepatocellular carcinoma 7721 cells,and the inhibitory effect was increased with the increase of drug concentration and duration of action.3.The above different concentrations of resveratrol acted on 7721 cells for 48 hours and thecell cycle determination results showed that:after 12.5,25,and 50?mol/L resveratrol treated 7721 cells for 48h,G₁ phase cells significantly increased,S phase cells significantly decreased.The G₂ phase cells significantly decreased in the dose group of 12.5 and 50?mol/L Resveratrol.4.The result of Hoechst 33258indicated that the cell apoptosis increased with the increase of resveratrol concentration.5.Annexin V-FITC/PI double staining method results showed that:The cell apoptosis rate increased with the increase of resveratrol concentration,the total apoptosis rate and early apoptosis were increased in the dose group of 50 and 100?mol/L Resveratrol.6.Western blot results showed that:different concentrations of resveratrol?12.5,25,50?mol/L?acting on 7721 cells for 48h,the expression of Bax,Caspase-3 and Caspase-9 protein was up-regulated,the expression of Bcl-2protein was down-regulated,the ratio of Bcl-2/Bax was declined.Conclusion:1.Resveratrol can inhibit the proliferation of hepatocellular carcinoma 7721cells,inducing apoptosis and arresting the cell cycle in G₁ phase.2.Resveratrol induced hepatocellular carcinoma 7721 cells apoptosis may be related to mitochondrial apoptosis pathway.