薯蓣皂苷经PI3K/AKT通路对肝癌Bel-7402细胞增殖和凋亡的影响

Effect of dioscin on proliferation and apoptosis of hepatocarcinoma cell line Bel-7402 via PI3K/AKT pathway

目的分析薯蓣皂苷对肝癌Bel-7402细胞增殖和凋亡的影响并探讨其机制。方法肝癌Bel-7402细胞分为空白组和薯蓣皂苷低、中、高剂量组(给予1、2、8μmol/L的薯蓣皂苷)及薯蓣皂苷+抑制剂组(给予8μmol/L的薯蓣皂苷+10μmol/L的磷脂酰肌醇3激酶/蛋白激酶B(PI3K/AKT)信号通路抑制剂LY294002),于处理后12、24、36、48及72 h时采用四甲基偶氮唑盐(MTT)比色法测定细胞活力,于24 h时采用流式细胞术检测细胞凋亡情况、采用蛋白免疫印迹法(Western blot)检测p-PI3K和p-AKT的表达。结果与空白组相比较,薯蓣皂苷低、中、高剂量组细胞活力及p-PI3K、p-AKT表达均下降,凋亡率升高(P<0.05),且各剂量组间两两比较,上述指标水平差异均有统计学意义(P<0.05);与薯蓣皂苷高剂量组比较,薯蓣皂苷+抑制剂组细胞活力及p-PI3K、p-AKT表达下降,凋亡率升高(P<0.05)。结论薯蓣皂苷可能通过抑制PI3K/AKT通路抑制肝癌Bel-7402细胞增殖,诱导Bel-7402细胞凋亡。

Objective To analyze the effect of dioscin on the proliferation and apoptosis of hepatocarcinoma cell line Bel-7402 and explore its mode of action.Methods Bel-7402 cells were divided into blank group,low-,medium-,and high-dose dioscin groups(given 1,2,8μmol/L dioscin,respectively)and dioscin+inhibitor group(given 8μmol/L dioscin+10μmol/L LY294002(phosphatidylinositol 3-kinase/protein kinase B(PI3K/AKT)signaling pathway inhibitor)).Tetramethylazolamide(MTT)colorimetry assay was used to detect cell viability at 12,24,36,48,and 72 hours after treatment,and cellular apoptosis was examined at 24 hours using flow cytometry.Western blot was used to detect the expressions of p-PI3K and p-AKT at 24 hours after treatment.Results When compared with blank group,cell viability and expressions of p-PI3K and p-AKT were significantly decreased,while the apoptosis rates were significantly increased in low-,medium-,and high-dose dioscin groups(P<0.05).Pairwise comparison among each dose group showed statistically significant differences in the levels of the above indicators(P<0.05).When compared with the high-dose dioscin group,cell viability and the expressions of p-PI3K and p-AKT in dioscin+inhibitor group were significantly decreased,while the apoptosis rate was significantly increased(P<0.05).Conclusion Dioscin might inhibit proliferation and induce apoptosis of Bel-7402 cells by inhibiting the PI3K/AKT pathway.