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目的探讨二甲双胍对大鼠催乳素瘤MMQ细胞增殖、细胞周期和凋亡的影响及其机制。方法采用(1.25、2.5、5、10、20)mmol/L二甲双胍处理MMQ细胞48 h,CCK-8法检测细胞增殖能力,流式细胞术检测细胞周期及细胞凋亡,Western blot法检测AMP活化的蛋白激酶α1/2(AMPKα1/2)、磷酸化的AMPKα(p-AMPKα)、哺乳动物雷帕霉素靶蛋白(m TOR)、磷酸化的m TOR(p-m TOR)、胰岛素样生长因子1受体(IGF-1R)、胞外信号调节激酶1/2(ERK1/2)、磷酸化的ERK1/2(p-ERK1/2)、蛋白激酶B(PKB/AKT)、磷酸化的AKT(p-AKT)、P21、细胞周期蛋白依赖激酶4(CDK4)、细胞周期蛋白D1(cyclin D1)、胱天蛋白酶3(caspase-3)、裂解的caspase-3(c-caspase-3)、Bcl-2、Bcl-2相关X蛋白(BAX)的变化。结果二甲双胍处理后,抑制MMQ细胞增殖,使细胞周期阻滞在G0/G1期,诱导细胞凋亡;细胞周期相关蛋白P21增加,CDK4、cyclin D1水平降低;c-caspase-3增加,caspase-3、Bcl-2降低;p-AMPKα水平增加、p-m TOR水平降低;IGF-1R、p-AKT及p-ERK水平降低。结论二甲双胍可能通过激活AMPK/m TOR通路、抑制IGF-1R通路从而抑制MMQ细胞增殖,诱导细胞周期阻滞及细胞凋亡。
Objective To investigate the effect and mechanism of metformin on proliferation, cell cycle and apoptosis of rat prolactinoid MMQ cells. Methods MMQ cells were treated with (1.25, 2.5, 5, 10, 20) mmol / L metformin for 48 h. Cell proliferation was measured by CCK-8 assay. Cell cycle and apoptosis were detected by flow cytometry. , Phosphorylated AMPKα (p-AMPKα), mammalian target of rapamycin (m TOR), phosphorylated m TOR (pm TOR), insulin-like growth factor 1 (IGF-1R), extracellular signal-regulated kinase 1/2 (ERK1 / 2), phosphorylated ERK1 / 2 (p-ERK1 / 2), protein kinase B (PKB / AKT), phosphorylated AKT p-AKT), P21, cyclinD1, caspase-3, c-caspase-3, Bcl- -2, Bcl-2-related protein X (BAX) changes. Results Metformin treatment inhibited the proliferation of MMQ cells and blocked the cell cycle in G0 / G1 phase and induced cell apoptosis. The expression of cell cycle related protein P21 increased, while the levels of CDK4 and cyclin D1 decreased. The expression of caspase-3 , Bcl-2 decreased; p-AMPKα level increased, pm TOR level decreased; IGF-1R, p-AKT and p-ERK levels decreased. Conclusion Metformin may inhibit the proliferation of MMQ cells and induce cell cycle arrest and apoptosis by activating AMPK / m TOR pathway and inhibiting IGF-1R pathway.