吲哚类化合物GY3通过AMPK/COX-2信号通路诱导乳腺癌细胞MCF-7凋亡

目的 探究吲哚类化合物GY3对人乳腺癌MCF-7细胞增殖和凋亡的作用,及其对腺苷酸活化蛋白激酶/环氧合酶-2(AMPK/COX-2)信号通路的影响.方法 以AMPK激活剂5-氨基4.甲酰胺咪唑核糖核苷酸(AICAR)和COX-2抑制剂塞来昔布为阳性对照药物,利用MTT方法、流式细胞术检测不同浓度的GY3、AICAR、塞来昔布及AICAR+塞来昔布联用对MCF-7细胞增殖和凋亡的影响;用Western blot方法检测不同浓度GY3和AMPK抑制剂Compound C联用对MCF-7细胞乙酰辅酶A羧化酶、磷酸化乙酰辅酶A羧化酶(P-ACC)、COX-2蛋白表达的影响.结果 GY3可上调P-ACC的表达、下调COX-2的表达(P＜0.01,P＜0.05),同时显著降低MCF-7细胞的活力(P ＜0.01,P＜0.05),抑制MCF-7细胞的增殖,诱导MCF-7细胞发生凋亡(P＜0.05),并且呈现一定的浓度依赖性;Compound C可有效地阻断GY3对AMPK的激活和COX-2的抑制作用,其作用与AIC-AR、塞来昔布及AICAR+塞来昔布联用的比较显示GY3的抑癌活性较高.结论 GY3能有效抑制乳腺癌细胞MCF-7的增殖,诱导其凋亡,其机制与激活AMPK、抑制COX-2的表达有关.

Indole compound GY3 induces cell apoptosis of breast cancer MCF-7 through AMPK/COX-2 pathway

Objective To investigate the effect of compound GY3 (GY3) on proliferation and apoptosis of breast cancer Michigan cancer foundation-7 (MCF-7) cells,and regulating downstream factors of adenosine 5'-monophosphate-activated protein kinase/cyclooxygenase-2 (AMPK/COX-2) signal pathway.Methods MTT assay was used to measure the inhibition rate of different concentration of GY3,5-minoimidazole-4-carboxyamide ribonucleoside (AICAR),celecoxib and AICAR combined with celecoxib on MCF-7 cells.Cell apoptosis rate was detected by FCM assay.The expressions of acetyl-coa carboxylase product of phosphorylation,phosphorylated acetyl-Co A carboxylase(P-ACC) and COX-2 were assayed in MCF-7 cells cultured with different concentration of GY3 and GY3 + Compound C by Western blot.Results GY3 inhibited MCF-7 cell growth in a concentration dependent manner (P ＜0.01,P ＜0.05).GY3 induced apoptosis of MCF-7 cells(P ＜0.05).GY3 could increase P-ACC and down regulate COX-2 expression(P ＜0.01,P ＜0.05).AMPK inhibitor Compound C could restore activation of AMPK and inhibition of COX-2 by GY3.Conclusion GY3 could inhibit COX-2 expression by activating AMPK,and subsequently induce apoptosis of MCF-7 cells.