ABSTRACT
Objective For Genistein has been reported to inhibit many tumors ,we investigate the role of autophagy in the proliferation inhibition to Hela cells by Genistein and the machanism of autophagy plays in this process.Methods Human cervical cancer Hela cells were divided into control group,Genistein group and 3-MA+Genistein group,the control group were cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum(FBS),Genistein group were cultured in various concentrations Genistein(25,50,100μmol/L),3-MA+Genistein group were treated with 5mmol/L 3-MA for 1h before cultured in 100μmol/L Genistein.The proliferation inhibitory rate of Hela cells was detected by MTT method.The ultrastructure changes of Hela cells was observed under transmission electronic microscope(TEM).The levels of autophagy-associated protein P62 and Beclin-1 were detected by Western blotting analysis.The expressions of autophagy-associated proteins LC3A/B in Hela cells were determined by fluorescent staining to analyse the autophagy induced by Genistein in Hela cells.Results Compared with control group ,the proliferation inhibitory rate of Hela cells was 20.9%±1.3%,33.5%±1.6% and 46.5%±3.2% when cultured in 25,50,100μmol/L Genistein(P<0.01).After treated with various concentrations Genistein for 48h, we observed a dose-dependent increase in the expression of Beclin-1 and decrease of P62.Confocal laser scanning microscopy confirmed the fluorescent density of LC3A/B expression in Hela cells treated with 100μmol/L Genistein increased significantly as compared with control group.TEM showed there are many vacuoles and double-membrane autophagosomes which involved cytoplasmic components in Hela cells treated with 100μmol/L Genistein.The proliferation inhibitory rate of Hela cells of Genistein group is decreased as compared with those in 3-MA+Genistein group[(46.5±3.2)% vs (58.2±2.2)%,P<0.01].Conclusion Genistein could inhibit Hela cells proliferation and induce autophagy.