蛇葡萄素对人宫颈癌SiHa细胞增殖、周期及凋亡的影响

目的探究蛇葡萄素(ampelopsin,AMP)对人宫颈癌SiHa细胞增殖、周期、凋亡的影响及其可能的作用机制。方法MTT法检测不同浓度(10、20、40、80、160、320μmol·L^-1)和不同干预时间(24、48、72 h)的AMP对SiHa细胞增殖抑制作用;Hoechst 33258法、Annexin-FITC/PI法检测AMP对SiHa细胞凋亡的影响;流式细胞术检测细胞周期变化;Rh-123法检测线粒体膜电位的变化;Western blot检测AMP对SiHa细胞中Bcl-2、Bax、Cleaved-caspase-3蛋白表达的影响。结果AMP对SiHa细胞抑制率呈浓度和时间依赖性(P<0.05),最低IC 50值为40.33μmol·L^-1;随着AMP浓度的增加或作用时间的延长,细胞凋亡率逐渐增加(P<0.05);当AMP浓度为80μmol·L^-1时,细胞周期阻滞在G 2/M期,呈一定的时间依赖性;线粒体膜电位随AMP浓度上升而下降;Bax/Bcl-2和Cleaved-caspase-3表达水平均上调,呈浓度和时间依赖性(P<0.05)。结论AMP明显抑制SiHa细胞的增殖,阻滞细胞周期于G 2/M期,可能通过线粒体途径诱导SiHa细胞凋亡。

Effects of ampelopsin on proliferation,cell cycle and apoptosis of human cervical cancer SiHa cells

Aim To investigate the effects of ampelopsin(AMP)on proliferation,cell cycle and apoptosis of human cervical cancer SiHa cells,and its possible mechanism of action.Methods MTT assay was used to detect the inhibitory effect of AMP with different concentrations(10,20,40,80,160,320μmol·L^-1)and different intervention time(24,48,72 h)on proliferation of SiHa cells.Hoechst 33258 staining and Annexin-FITC/PI kit were used to detect the effect of AMP on apoptosis of SiHa cells.Flow cytometry was used to detect cell cycle.Rh-123 staining was employed to detect the changes of mitochondrial membrane potential.Western blot was used to detect the effect of AMP on the expression of Bcl-2,Bax,cleaved-caspase-3 proteins in SiHa cells.Results The inhibitory rate of AMP on SiHa cells was concentration-and time-dependent(P<0.05),and the smallest IC 50 value was 40.33μmol·L^-1.The apoptotic rate increased with the rise of AMP concentration or the prolongation of action time(P<0.05).When AMP concentration was 80μmol·L^-1,cell cycle was blocked in G 2/M phase with time-dependence.Mitochondrial transmembrane potential decreased with the rise of AMP concentration.The expression levels of Bax/Bcl-2 and cleaved-caspase-3 were both up-regulated in a concentration-and time-dependent manner(P<0.05).Conclusion AMP can significantly inhibit the proliferation of SiHa cells and block cells at G 2/M phases,and its mechanism may be via inducing apoptosis of SiHa cells through mitochondrial pathway.