目的 研究黃芪有效成分芒柄花素與黃芪甲苷IV對人食道癌HCE-4細胞的凋亡作用及其分子機制。方法 MTT法檢測黃芪提取物(陽性對照藥)、芒柄花素與黃芪甲苷IV對人食道癌HCE-4細胞的增殖抑制作用,Annexin V-FITC/PI雙染法檢測3種藥物誘導(dǎo)HCE-4細胞凋亡的能力,Western blotting法檢測凋亡相關(guān)蛋白的表達變化。結(jié)果 芒柄花素與黃芪甲苷IV對HCE-4細胞具有良好的生長抑制作用,其效果呈濃度依賴性;芒柄花素與黃芪甲苷IV能夠誘導(dǎo)HCE-4細胞的凋亡且呈時間依賴性;芒柄花素與黃芪甲苷IV處理HCE-4細胞后,抗凋亡蛋白p-AKT的表達量明顯減少,促凋亡蛋白cleaved-caspase-3表達量增加。結(jié)論 黃芪有效成分芒柄花素與黃芪甲苷IV通過調(diào)控AKT信號轉(zhuǎn)導(dǎo)途徑誘導(dǎo)人食道癌HCE-4細胞凋亡。

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.

黑龍江省大學(xué)生創(chuàng)新創(chuàng)業(yè)訓(xùn)練計劃項目(201510223003);黑龍江省自然基金資助項目(LC2015036);黑龍江省博士后科研啟動項目(LBH-Q13132);學(xué)成、引進人才科研啟動計劃項目(XYB2013-24)