Study On Compound Kushen Injection On Promoting Apoptosis Induced By Autophagy In Bladder Cancer Cells

Objective:The influence of Compound Kushen Injection on bladder cancer cells was explored through the cell culture experiment in vitro,and the possible molecular mechanism was preliminarily explored,providing theoretical basis for the clinical treatment of bladder cancer by Compound Kushen Injection Method:CCK-8 method was used to detect the effect of Compound Kushen Injection on the proliferation of bladder cancer cells and the synergistic effect with epirubicin;cell scratch test was used to observe the effect of Compound Kushen Injection on the migration of bladder cancer cells;CCK-8 method was used to detect the effect of apoptosis inhibitor,necrosis inhibitor and autophagy inhibitor on the lethal effect of Compound Kushen Injection on bladder cancer cells;flow cytometry was used to detect the effects of Compound Kushen Injection on apoptosis and cycle of bladder cancer cells;MDC staining on autophagy of bladder cancer cells;RT-qPCR?WB and immunofluorescence were used to detect the effects of Compound Kushen Injection on apoptosis key genes,apoptosis key proteins,autophagy key genes and autophagy key proteins of bladder cancer cells.Results:Compound Kushen Injection can effectively inhibit the proliferation of bladder cancer T24 cells in a dose-dependent manner,and the inhibition rate of cell growth can increase with the increase of drug concentration.The combination of Compound Kushen Injection and epirubicin at a certain time and concentration has a synergistic effect.The changes of cell morphology and the increase of dead cells were observed 24 and 48 hours after the intervention of Compound Kushen Injection on bladder cancer cells.The results of CCK-8showed that apoptosis inhibitor and autophagy inhibitor could reduce the lethal effect of Compound Kushen Injection on T24 cells,while programmed necrosis inhibitor had no significant effect.The results of flow cytometry showed that compared with the control group,the Compound Kushen Injection could significantly induce apoptosis of bladder cancer cells,and had a dose-dependent effect.The combination of the Compound Kushen Injection and epirubicin had a better effect on apoptosis of bladder cancer T24 cells.With the increase of the concentration of Compound Kushen Injection,G2 phase gradually increased and cell cycle was blocked.MDC staining showed that Compound Kushen Injection could induce autophagy of bladder cancer cells.The results of RT-qPCR ? Western blotting and immunofluorescence showed that with the increase of the concentration of Compound Kushen Injection,the expression of Bcl-2 gene and protein decreased,the expression of Bax,Caspase-3 gene and protein increased,the ratio of Bcl-2/Bax decreased,the expression of Beclin 1,LC3?,gene and protein increased,P62 gene and protein decreased.Conclusion:Compound Kushen Injection can inhibit the proliferation and migration of bladder cancer T24 cells.Compound Kushen Injection can cause G2 arrest of bladder cancer cells,induce autophagy of bladder cancer cells and promote apoptosis of bladder cancer cells.