The Effects Of Lycorine On The Clone Formation,Apoptosis And Cell Cycle Of Hepatocellular Carcinoma Cells In Vitro

Objective:1.To investigate the effect of Lycorine on the cell viability of Hepatocellular Carcinoma Bel-7402 and SNU-423 cells.2.To investigate the anti-cancer effects of Lycorine on the clone formation,apoptosis and cell cycle of Hepatocellular Carcinoma Bel-7402 and SNU-423 cells.3.To explore the possible molecular mechanism of Lycorine affecting clone formation,apoptosis and cell cycle of the Hepatocellular Carcinoma Bel-7402 and SNU-423 cells.Methods:1.MTT assay was used to examine the effect of Lycorine on the cell viability of the Hepatocellular Carcinoma Bel-7402 and SNU-423 cells after treatment at 24 h,48h and 72 h.2.For the Hepatocellular Carcinoma Bel-7402 and SNU-423 cells,the effects of Lycorine on clone formation,and apoptosis were assessed with clone formation assay and flow cytometry,respectively.Flow cytometry was used to examine the effect of Lycorine on the cell cycle of Hepatocellular Carcinoma Bel-7402 and SNU-423 cells.3.Western blot was performed to explore the possible molecular mechanism of Lycorine affecting clone formation,apoptosis and cell cycle of the Hepatocellular Carcinoma Bel-7402 and SNU-423 cells.Results:1.The MTT assay result showed that the relative viability of the Hepatocellular Carcinoma Bel-7402 and SNU-423 cells was decreased after the treatment of Lycorine compared with the control group(P<0.05).2.Compared with the control group,Lycorine suppressed the clonogenic ability of the Hepatocellular Carcinoma Bel-7402 and SNU-423 cells in a dose-dependent manner(P<0.05),according to the results of clone formation assay.Compared with the control group,Lycorine also induced apoptosis of the Hepatocellular Carcinoma Bel-7402 and SNU-423 cells in a concentration-dependent manner(P<0.05),according to the results of flow cytometry.According to the results of flow cytometry,Lycorine increased the proportion of G2/M phase cells of the Hepatocellular Carcinoma Bel-7402 and SNU-423 cells(P<0.05),compared with the control group.3.Western blot results showed that Lycorine could down-regulate the expression of CyclinA,cdc2 and p-Akt proteins(P<0.05),while up-regulate the level of Cleaved Caspase-3 protein(P<0.05).Conclusion:1.Lycorine can effectively reduce the relative viability of the Hepatocellular Carcinoma Bel-7402 and SNU-423 cells in a time-dependent manner.2.Lycorine inhibits the clonogenic ability of the Hepatocellular Carcinoma Bel-7402 and SNU-423 cells in a dose-dependent manner,apoptosis of Hepatocellular Carcinoma Bel-7402 and SNU-423 cells was induced,and cell cycle of the Hepatocellullar Carcinoma Bel-7402 and SNU-423 cells was arrested at G2/M phase.3.Lycorine may inhibit the proliferation of the Hepatocellular carcinoma Bel-7402 and SNU-423 cells by down-regulating the expression of Cyclin A and cdc2,up-regulating the expression of Cleaved Caspase-3,meanwhile inhibit the phosphorylation of Akt,eventually inhibit the proliferation of the Hepatocellular Carcinoma Bel-7402 and SNU-423 cells.