大蒜素诱导人子宫内膜癌Ishikawa细胞凋亡的作用机制研究

Study on the Mechanism of Allicin-induced Apoptosis in Ishikawa Cells of Human Endometrial Carcinoma

目的:探讨大蒜素诱导人子宫内膜癌Ishikawa细胞凋亡的作用机制。方法:分别以二甲基亚砜(空白对照组)、大蒜素(12.5、25、50μg/mL)和LY2940025μg/mL干预对数生长期Ishikawa细胞。48 h后,用CCK-8法检测细胞增殖抑制率,流式细胞术分析细胞凋亡状况,Western Blotting法检测p-PI3K、p-AKT、Cleaved Caspase-3、Bcl-2、Bax蛋白表达。结果:与空白对照组比较,经大蒜素12.5、25、50μg/mL或LY2940025μg/mL干预能够提高Ishikawa细胞增殖抑制率和凋亡率;经大蒜素25、50μg/mL或LY2940025μg/mL干预能够下调p-PI3K、p-AKT、Bcl-2表达并上调Cleaved Caspase-3、Bax表达,提高Bax/Bcl-2比值,差异有统计学意义(P<0.05或P<0.01)。与LY294002组比较,经大蒜素50μg/mL干预能够提高Ishikawa细胞增殖抑制率和凋亡率,下调p-PI3K、p-AKT表达并上调Cleaved Caspase-3、Bax表达,提高Bax/Bcl-2比值,差异有统计学意义(P<0.05或P<0.01)。结论:大蒜素能够诱导人子宫内膜癌Ishikawa细胞凋亡,可能与抑制PI3K/AKT信号通路活化而使其下游促凋亡蛋白表达上调有关。

Objective:To investigate the effects and mechanism of Allicin on apoptosis of Ishikawa cells in human endometrial carcinoma.Methods:Ishikawa cells in logarithmic growth phase was treated with DMSO(blank control group),Allicin(12.5,25,50μg/mL)and LY2940025μg/mL.48 h later,the cell proliferation inhibition rate was detected by CCK-8 method,cell apoptosis was analyzed by flow cytometry,and protein expression of p-PI3K,p-AKT,Cleaved Caspase-3,Bcl-2,and Bax were detected by Western blotting.Results:Compared with the blank control group,intervention of Allicin 12.5,25,50μg/mL and LY2940025μg/mL could increase Ishikawa cells proliferation inhibition rate and apoptosis rate,down-regulate the expression of p-PI3K,p-AKT,Bcl-2 and up-regulate the expression of Cleaved Caspase-3,Bax,increase the ratio of Bax/Bcl-2,all of which were statistically significant(P<0.05 or P<0.01).Compared with the LY2940025μg/mL group,intervented by Allicin 50μg/mL could increase Ishikawa cells proliferation inhibition rate and apoptosis rate,down-regulate the expression of p-PI3K,p-AKT and up-regulate the expression of Caspase-3,Bax,increase the ratio of Bax/Bcl-2,all of which were statistically significant(P<0.05 or P<0.01).Conclusion:Allicin can induce apoptosis of human endometrial cancer Ishikawa cells,which may be related to the inhibition of PI3K/AKT signaling pathway activation and up-regulation of downstream pro-apoptotic proteins.