Wnt信号通路在姜黄素诱导乳腺癌MCF-7细胞凋亡中作用机制

目的 探讨Wnt通路在姜黄素诱导人乳腺癌MCF-7细胞凋亡中的作用机制.方法 将MCF-7细胞培养液随机分为5组,分别加入15、30、60、120μmol/L的姜黄素,对照组不加,培养12,24,48 h后采用CCK-8法检测细胞增殖能力.分A、B两组(剂量组、时间组)采用Annexin V-FITC/PI双染流式细胞术(FCM)观测细胞凋亡及细胞周期分布;Western Blotting法检测A、B两组的Wnt通路相关蛋白,并进行相应的相关性分析探讨Wnt通路与乳腺癌MCF-7细胞凋亡的关系.结果 CCK-8结果显示,随培养时间延长,姜黄素各浓度组MCF-7细胞增殖抑制率逐渐增高(均P<0.05);在同一时间点,随着姜黄素浓度升高,细胞增殖抑制率逐渐增高(均P<0.05).FCM染色结果显示,随着姜黄素浓度增加、作用时间延长MCF-7细胞凋亡指数逐渐上升(均P<0.05),且G1/S期细胞增加越多(P<0.05).Western Blotting实验结果显示MCF-7细胞中Wnt1、β-catenin的表达下调程度呈现剂量时间依赖性(P<0.05).相关性分析显示细胞抑制率、G0/G1期细胞百分比、细胞凋亡率均与Wnt通路相关蛋白Wnt1、β-catenin的表达呈现较明显的相关性(P<0.05).结论 姜黄素的抗癌活性与抗增殖能力具有明显的剂量时间依赖性,且Wnt通路在姜黄素诱导乳腺癌MCF-7细胞抑制、凋亡过程中发挥了重要作用.

Role of Wnt Signaling Pathway in Curcumin-induced Apoptosis of Breast Cancer Cells MCF-7

Objective To investigate the mechanism of Wnt pathway in curcumin-induced apoptosis of human breast cancer cells MCF-7.Methods The MCF-7 cells were randomly divided into 5 groups,respectively received 15,30,60 and 120μmol/L curcumin,and the control group,CCK-8 cells were cultured for 12,24 and 48 hours to detect proliferative capacity . Apoptosis and cell cycle distribution were observed by Annexin V-FITC/PI double staining flow cytometry (FCM).The Wnt path-way of group A and group B was detected by western blotting method .And to explore the relationship between the Wnt pathway and the apoptosis of breast cancer MCF-7 cells.Results CCK-8 showed that the proliferation inhibition rate of MCF-7 cells in-creased gradually with the increase of curcumin concentration (P<0.05);at the same time point,with the increase of curcumin concentration,the inhibition of cell proliferation (P<0.05).FCM staining showed that the apoptotic index of MCF-7 cells in-creased gradually with the increase of curcumin concentration (P<0.05) and the increase of G1/S phase cells (P<0.05). Western blotting results showed that the expression of Wnt 1 and β-catenin in MCF-7 cells was dose and time dependent ( P<0.05).The correlation analysis showed that the percentage of cells ,G0/G1 phase, apoptosis rate were closely correlated with the expression of Wnt1 andβ-catenin (P<0.05).Conclusion The anticancer activity and antiproliferative ability of curcumin have a dose and time dependent manner ,and Wnt pathway has a significant effect on the expression of Wnt 1 and β-catenin in curcu-min-induced breast cancer MCF-7 cells,and it plays an important role in the process of apoptosis .