Effects on apoptosis and cell cycle of RA in HT29 cells / 实用医学杂志

ABSTRACT

Objective To investigate the effects of Raddeanin A(RA)on apoptosis and cell cycle of hu-man colon cancer HT29 cells,and discuss the possible mechanism.Methods Different concentration of RA 1,2, 4,8,16μmol/L disposed HT29 cells 12,24,48 hour,3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bro-mide(MTT)assay was used for the cell proliferation;Hoechst33258 staining was performed to observe the effects of RA on apoptosis morphology;Flow cytometry(FCM)was devoted to calculate apoptosis rate and detect cell cycle;The expression of Cleaved-caspase-3,Cleaved-PARP,CyclinB,CDK1 proteins and other proteins were tested by Western blot(WB). Results RA could inhibit the proliferation of HT29 cells,and associated with concentration and time(P<0.05);The IC50of 12 h,24 h,48 h were(9.31 ± 0.63)μmol/L,(4.88 ± 1.02)mol/L,(3.60 ± 0.89) μmol/L.The nucleus pycnosis,fracture were detected by Hochest33258 staining,RA induced apoptosis in HT29 cells;Western blot(WB)assay showed that the expression of Cleaved-caspase-3,Cleaved-PARP rised.HT29 cells were arrested at the period of G2/M,and the CyclinB,CDK1 proteins down-regulated.WB result showed that PI3K, AKT proteins up-regulated,while p-PI3K,p-AKT protein down-regulated.Conclusion RA can inhibit the prolifer-ation of colon cancer HT29 cells and its mechanism is probably by regulating the PI3K/AKT signaling pathways which through inducing apoptosis and blocking the cell cycle.