麦冬皂苷B通过抑制PI3K/Akt/mTOR通路诱导人黑色素瘤A375细胞凋亡

目的探讨麦冬皂苷B对人黑色素瘤(Melanoma)A375细胞增殖,凋亡和细胞周期的影响以及相关机制。方法麦冬皂苷B处理A375细胞后,采用CCK-8法检测细胞活力;流式细胞仪检测A375细胞周期变化及细胞凋亡水平;Hoechst33258荧光染色法观察细胞凋亡形态;免疫印迹法检测麦冬皂苷B对A375细胞PI3K,p-PI3K,AKT,p-AKT蛋白以及通路下游凋亡相关蛋白Bcl-2,Bax,Cleaved-caspase3表达的影响。结果麦冬皂苷B能抑制A375细胞增殖活力及PI3K/Akt/mTOR通路的活化;麦冬皂苷B促进了A375细胞内Bax和Cleaved-caspase 3的表达,抑制Bcl-2表达,诱导人黑色素瘤细胞发生凋亡;麦冬皂苷B阻滞了A375细胞细胞周期,G0/G1期细胞比例明显升高;PI3K/Akt/mTOR通路活化剂IGF-1处理A375细胞后抑制了麦冬皂苷B的上述作用。结论麦冬皂苷B能抑制A375细胞的增殖,诱导细胞发生凋亡,阻滞细胞周期于G0/G1期。

Ophiopogonin B Induces the Apoptosis of Melanoma A375 Cells by Suppressing PI3K/Akt/mTOR Signaling Pathway

Objective In order to investigate the effects of Ophiopogonin B on the proliferation,apoptosis and cell cycle in human melanoma A375 cells line via PI3 K/Akt/mTOR signal pathway.Methods The cell growth was determined by CCK-8 assay.The effects of Ophiopogonin B on the cell cycle and apoptosis were detected by flow cytometry with annexin V-FITC/PI.Apoptotic cell morphology was visualized by Hoechst 33258 staining.The protein expressions were examined by western blotting analysis.Results Ophiopogonin B inhibited the melanoma A375 cell viability in a dose-dependent manner.Ophiopogonin B suppressed the activation of PI3 K/Akt/mTOR signal pathway.Ophiopogonin B induced apoptosis of A375 cells through reducing the expression of Bcl-2 and enhancing the expression of Bax and Cleaved-caspase-3.Morphological changes were seen in apoptotic A375 cells by Hoechst 33258 staining.Ophiopogonin B treatment resulted in an arrest of cell cycle at G0/G1 phase.However,treatment with insulin-like growth factor 1(IGF-1)reversed these Ophiopogonin B-mediated effects on the activation of PI3 K/Akt/mTOR signal pathway,cell apoptosis and cell cycle.Conclusion Ophiopogonin B inhibits cell proliferation,triggers apoptosis and blocks cell cycle in human melanoma A375 cells via inhibiting PI3 K/Akt/mTOR signal pathway.