YAP蛋白磷酸化参与二甲双胍抑制乳腺癌MCF-7细胞生长

YAP Protein Phosphorylation in Metformin Inhibits Growth of MCF-7 Cells in Breast Cancer

目的验证二甲双胍在人乳腺癌MCF-7细胞中的细胞增殖的影响,并探讨二甲双胍在乳腺癌中介导的抗肿瘤活性的潜在分子靶标。方法使用含或不含二甲双胍的培养基MCF-7细胞,使用血细胞计数器进行计数。通过CCK-8方法检测不同浓度的二甲双胍对细胞增殖的影响,并获取半数抑制浓度(IC50),并以此浓度进行后续实验研究。使用实时荧光定量Real-time PCR检测目的基因的表达。使用蛋白印迹法检测目的蛋白LATS1、STK4、YAPl、磷酸化YAPl(p-YAPl)的表达情况。结果二甲双胍呈剂量依赖性抑制MCF-7细胞的增殖,组间比较差异有统计学意义(P<0.05)。二甲双胍抑制MCF-7细胞增殖IC50浓度为40 mmol/L。二甲双胍处理组细胞YAP、LATS1、SKT4的mRNA表达上调,YAPl蛋白表达量下调,p-YAPl蛋白表达量升高,提示YAP被磷酸化。结论二甲双胍可抑制人乳腺癌MCF-7细胞的增殖,且抑制率随浓度及时间增加。其机制可能与YAP蛋白磷酸化有关。

Objective To verify the effect of metformin on cell proliferation in MCF-7 cells of human breast cancer,and to explore the potential molecular target of metformin in breast cancer mediated anti-tumor activity.Methods A medium MCF-7 cells containing or not containing metformin were used to count the blood cell counters.The effects of different concentrations of metformin on cell proliferation were detected by CCK-8 method,and half of the inhibition concentration(IC50)was obtained,and the subsequent experimental study was carried out with this concentration.Real-time fluorescence quantitative Real-time PCR was used to detect the expression of target genes.The expression of target protein LATS1,STK4,YAPl and phosphorylation YAPl(p-YAPl)was detected by protein imprinting.Results Metformin was dose-dependent to inhibit the proliferation of MCF-7 cells,and the differences between the groups were statistically significant(P<0.05).Metformin inhibited the proliferation of MCF-7 cells and measured IC50 concentrations of 40 mmol/L,metformin treatment group cells YAP,LATS1.The expression of mRNA in SKT4 was increased,and the expression of YAPl protein in metformin treatment group decreased p-YAPl protein expression,suggesting that YAP was phosphorylated.Conclusion Metformin can inhibit the proliferation of MCF-7 cells in human breast cancer,and the inhibition rate increases with concentration and time.The mechanism may be related to the phosphorylation of YAP protein.