Study On The Effects And Molecular Mechanisms Of Matrine On The Proliferation And Migration Of Hepatocellular Carcinoma HepG-2 Cells

Objective:To investigate the effects and molecular mechanisms of matrine(MAT)on the proliferation and migration of hepatocellular carcinoma cell line HepG-2.Methods:HepG-2 cells were treated with different concentrations of MAT,1.The effect of MAT on the proliferation of HepG-2 cells were detected by MTT assay;2.The effect of MAT on the colony forming ability of HepG-2 cells were detected by plate cloning assay;3.The effect of MAT on the migration of HepG-2 cells were detected by wound-healin and Transwell chamber assay;4.Hoechst 33342 staining were used to observed that the effect of MAT on apoptosis of HepG-2 cells;5.The expression lever of Bax,Bcl-2,Ezrin,p-Ezrin and EMT related proteins(E-cadherin,Vimentin and Snail)were detected by Western blot assay.Results:1.HepG-2 cells were treatment with 0.25,0.5,1.0,1.5 and 2.0mg/ml of MAT and the proliferation ability of detected by MTT assay,the results showed that:the proliferation ability of HepG-2 cells were significantly lower than the control group,which showed in a dose-and time-dependent manner(P<0.01);2.The colony-forming ability of HepG-2 cells were reduced after exposure to MAT(0.5,1.0 and 1.5mg/ml)in a dose dependent manner compared with control group(P<0·01);3.The result of Hoechst 33342 staining showed that:the apoptosis of HepG-2 cells were significantly increased,compared with the control group after treatment with 0.5,1.0 and 1.5mg/ml of MAT,and showed in dose-dependent manner(P<0.01);4.The wound-healing and Transwell chamber assay results showed that:the migration ability of HepG-2 cells were significantly inhibited after treated with 0.5,1.0 and 1.5mg/ml of MAT,and showed in dose-dependent manner(P<0.01);5.The Western blot showed that:HepG-2 cells were treated with 0.5,1.0 and 1.5mg/ml of MAT,the expression of epithelial marker E-cadherin protein and the ratio of Bax/Bcl-2 were increased(P<0.01),and the expression of Ezrin,p-Ezrin,Vimentin and snail protein were down regulated(P<0.05 or P<0.01).Conclusion:1.MAT inhibited the proliferation and colony formation of HepG-2 cells,which mechanism may be related to the apoptosis of HepG-2 cells,induced by MAT;2.The migration of HepG-2 cells in both horizontal and vertical direction were inhibited by MAT,and the reversal of EMT may be involved in via regulating the expression of Ezrin and EMT related proteins.