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[摘要]
目的 研究黄芪有效成分芒柄花素与黄芪甲苷IV对人食道癌HCE-4细胞的凋亡作用及其分子机制。方法 MTT法检测黄芪提取物(阳性对照药)、芒柄花素与黄芪甲苷IV对人食道癌HCE-4细胞的增殖抑制作用,Annexin V-FITC/PI双染法检测3种药物诱导HCE-4细胞凋亡的能力,Western blotting法检测凋亡相关蛋白的表达变化。结果 芒柄花素与黄芪甲苷IV对HCE-4细胞具有良好的生长抑制作用,其效果呈浓度依赖性;芒柄花素与黄芪甲苷IV能够诱导HCE-4细胞的凋亡且呈时间依赖性;芒柄花素与黄芪甲苷IV处理HCE-4细胞后,抗凋亡蛋白p-AKT的表达量明显减少,促凋亡蛋白cleaved-caspase-3表达量增加。结论 黄芪有效成分芒柄花素与黄芪甲苷IV通过调控AKT信号转导途径诱导人食道癌HCE-4细胞凋亡。
[Key word]
[Abstract]
Objective To investigate the pharmacologic effects and apoptotic mechanism of active components from Astragalus membranaceus on human esophageal cancer HCE-4 cells. Methods The viabilities of HCE-4 cells were measured by MTT assay. The inhibition of active components from A. membranaceus on apoptosis of HCE-4 cells was detected by Annexin V-FITC/PI double staining. The apoptotic-related protein expression levels were determined by Western blotting. Results Formononetin and astragaloside IV suppressed the proliferation of HCE-4 cells in a dose-dependent manner. The Annexin V-FITC/PI double staining results showed that formononetin and astragaloside IV could induce HCE-4 cells apoptosis in a time-dependent manner. The Western blotting results showed that formononetin and astragaloside IV could significantly down-regulate p-AKT, Pro-caspase-9, and Pro-caspase-3 protein expression in HCE-4 cells. Conclusion Active components from A. membranaceus such as formononetin and astragaloside IV significantly inhibits the proliferation of human esophageal cancer HCE-4 cells by inducing mitochondrial dependent apoptosis via AKT signaling pathway.
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[基金项目]
黑龙江省大学生创新创业训练计划项目(201510223003);黑龙江省自然基金资助项目(LC2015036);黑龙江省博士后科研启动项目(LBH-Q13132);学成、引进人才科研启动计划项目(XYB2013-24)