The Effects Of Icariin On Proliferation And Apoptosis Of Hepatocellular Carcinoma HepG2 Cells

Objectives: To observe the effects of icariin(ICA)on the proliferation,apoptosis and cell cycle of Hep G2 human hepatoma cells and to explore its mechanism of action.Methods: Different concentrations of icariin(0,12.5,25,50,100,200 ?g/m L)were used to intervene Hep G2 cells.The cell viability was observed under inverted microscope.The cell proliferation was determined by MTT assay at 24,48,and 72 h.Flow cytometry was used to analyze the changes of cell cycle after 48 h,Hoechst 33258 staining was used to observe the apoptosis after 48 h,Annexin V-FITC/PI double staining was used to analyze the change of apoptosis rate after 48 h,and Western blot was used to detect the changes.After treatment with different concentrations of icariin(0,25,50,and 100 ?g/m L)for 48 h,the expressions of apoptotic signalling factors Caspase-3,Caspase-9,Bcl-2,and Bax protein were changed;SPSS21.0 software One-way analysis of variance between groups,LSD analysis between the two groups,P <0.05 was considered statistically significant.Results: 1.Icariin inhibits the proliferation of human hepatocellular Hep G2 cells: MTT results show that icariin can significantly inhibit the proliferation of Hep G2 cells at 12.5,25,50,100,and200 ?g/m L,which is statistically different from that of the control group(P<0.05),and with increasing concentration and time extension,the inhibition was enhanced(P<0.05).2.Epimedium can induce Hep G2 cell cycle arrest: Flow cytometry results after PI staining indicated that 25,50,and 100 ?g/m L of icariin interfered with Hep G2 cells for 48 h,and cell cycle G0/G1 cells increased significantly.(P<0.05),cells treated with 50 and 100 ?g/m L significantly decreased in S phase and G2/M phase cells(P<0.05).3.Epimedium can induce apoptosis in Hep G2 cells: after48 hours of Hep G2 cells treated with icariin at 12.5,25,50,100,and 200 ?g/m L,staining with Hoechst 33258 showed that with increased drug concentration,Apoptosis significantly changed;the total cell apoptosis rates of icariin at 25,50,and 100 ?g/m L for 48 h were 6.33%±0.78%,16.06%±1.22%,and 22.38%±1.83%,respectively.Compared with the control group,there was a statistical difference(P<0.05),and with the increase of dose,the apoptosis increased(P<0.05).4.Western blot: After 25,50,and 100 ?g/m L icariin treatment of Hep G2 cells for 48 h,the results showed that the pro-apoptotic protein Bax was up-regulated,the expression of anti-apoptotic protein Bcl-2 was down-regulated,and the ratio of Bax/Bcl-2 increased.Compared with the control group,the difference was statistically significant(P<0.05);the expression of apoptosis signalling factors Caspase-3 and Caspase-9 was up-regulated,and there was statistical difference between 50 and 100 ?g/m L compared with the control group(P<0.05).Conclusions:1.Icariin can significantly inhibit Hep G2 cell proliferation,induce apoptosis,and arrest cell cycle in G0/G1 phase.2.Icariin can induce apoptosis of Hep G2 cells,promote the expression of Bax/Bcl-2 protein and up-regulate the expression of Caspase-3 and Caspase-9,suggesting that icariin can induce Hep G2 apoptosis and mitochondrial apoptosis.The approach is closely related.