【摘要】 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。更多还原

【Abstract】 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 and120μmol / 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 pathway 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 increased 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 increased 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 Wnt1 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 and β-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 Wnt1 and β-catenin in curcumin-induced breast cancer MCF-7 cells,and it plays an important role in the process of apoptosis.更多还原