去甲斑蝥素通过Wnt/β-catenin信号通路诱导乳腺癌细胞MCF-7凋亡

Apoptosis in breast cancer cells MCF-7 induced by norcantharidin through the Wnt/β-catenin signaling pathway

目的:考察去甲斑蝥素(NCTD)对乳腺癌细胞MCF-7的凋亡诱导作用,并对其机制进行研究。方法:采用MTT法考察不同浓度NCTD (1 000、500、250、125、62.5、31.3、0μmol/L)对MCF-7细胞成活率的影响并计算IC50值;采用PI染色的流式细胞术考察不同浓度NCTD (0、50、100μmol/L)对MCF-7细胞周期的影响;采用Annexin V-FITC、PI双染法的流式细胞术考察不同浓度NCTD (0、50、100μmol/L)对MCF-7细胞的凋亡诱导作用;采用Western blot法考察不同浓度NCTD (0、25、50、100μmol/L)对MCF-7细胞中Wnt/β-catenin信号通路相关蛋白的影响。结果:NCTD可浓度依赖地降低MCF-7细胞的成活率(P<0.01),IC50值为(100.73±3.17)μmol/L;NCTD可浓度依赖地降低MCF-7细胞处于G0/G1期细胞百分比和升高G2/M期的细胞百分比(P<0.01);100μmol/L NCTD升高MCF-7细胞中凋亡细胞的百分比(P<0.01);NCTD还可显著诱导MCF-7细胞中β-catenin及其下游蛋白MMP-7的表达水平下降。结论:NCTD可通过Wnt/β-catenin通路诱导MCF-7细胞发生凋亡。

Objective: To investigate the apoptotic effect of norcantharidin(NCTD) in breast cancer cells MCF-7 and elucidate its underlying mechanisms. Methods: The MCF-7 cells were treated with different concentrations of NCTD(1 000, 500, 250, 125, 62.5,31.3, 0 μmol/L) for 72 h. Cell viabilities were examined by MTT assay. The cell cycle analysis was examined by using FACS after PI staining. FACS after AnnexinV-PI staining was also used to examine the apoptotic cell rates. The expression level of Wnt/β-catenin pathway related proteins was determined by Western blot. Results: NCTD could concentration-dependently decrease the cell viabilities of MCF-7 cells, IC50 was(100.73±3.17) μmol/L. It can also decrease the percentage of G0/G1 phase cells and increase the percentage of G2/M phase cells. NCTD could induce apoptosis in MCF-7 cells and also significantly decrease the expression level ofβ-catenin and its downstream protein MMP-7. Conclusion: NCTD can induce apoptosis in MCF-7 cells through the Wnt/β-catenin signaling pathway.