Pseudolaric Acid B Induced Cell Cycle Delay And Apoptosis In Human Hepatocellular Carcinoma SMMC7721 Cells

Purpose:PAB?Pseudolaric acid B?inhibited tumor cell proliferation,but whether PAB had broad effect on HCC was not clear.Therefore this study detected the role of PAB on HCC SMMC7721,which would be important for development of PAB for anti-HCC.Method:MTT methods were used to determine the growth of PAB-treated cells;Tubulin aggregation induced by PAB was detected by fluorescence staining;Cell immigration was detected by scratching line;Western blot was used to detect protein expression;Flow cytometry detected the cell cycle distribution;Gene transcription of CyclinB1 and CDK1 was detected by Real time PCR after PAB treatment;DNA fragmentation was determined by agarose gel electrophoresis.Results:It was found that PAB inhibited SMMC7721 cell growth in time-and dose-dependent manner through intervening the tubulin of SMMC7721 cells.PAB inhibited cell immigration.Meanwhile PAB promoted tetraploid cells accumulation from 24 h to 48 h,and induced cell apoptosis at 48 h,while together with CDK1 inhibitor,apoptosis occured at24 h.Further study proved that apoptosis-induced by PAB at 48 h was related to the expression of MAPK family member and Caspase-3,Caspase-9 and Caspase-8.And at 24 h,PAB together with CDK1 inhibitor induced apoptosis because they decreased the expression of MAPK proteins and activated Caspase-3.And cell cycle related protein cyclinB1 and CDK1 expression and transcription further confirmed that PAB induced mitotic delay at early time and mitotic slippage into G₁ phase at late time,and other cell cycle related protein expression also support the point.Conclusion:Therefore PAB had anti-SMMC7721 role,and together with CDK1inhibitor it is more efficient.