二甲双胍通过诱导G1期阻滞抑制骨肉瘤细胞增殖

The inhibition effect of metformin on proliferation in osteosarcoma MG63 cell line by G1 phase blocking

目的：检测二甲双胍对人骨肉瘤细胞株MG63增殖的影响。方法：不同浓度二甲双胍刺激MG63-MTT法检测细胞体外增殖能力：5mmol／L二甲双胍作用48h，流式细胞术检测MG63细胞周期分布．免疫印迹检测cyclinD1表达以及AMPK、AKT、S6K及s6蛋白磷酸化水平。结果：0～50mmol／L的二甲双胍对MG63细胞增殖的抑制作用，除10mmol／L和20mmol／L两组间外，其他处理组间均表现出显著性差异（P〈0．01）。5mmol／L二甲双胍诱导MG63细胞发生G1期阻滞（P〈0．01），抑制cylinD1、P-S6K1及P-s6表达，促进AMPK和AKT的磷酸化。结论：二甲双胍通过诱导细胞周期G1期阻滞．通过激活AMPK和AKT的磷酸化抑制roTOR通过抑制MG63增殖。

Objective To explore the effect of metformin on proliferation activity in osteosarcoma MG63 cell line. Methods MG63 cells were stimulated by metformin at different concentrations and the proliferation activities were detected by Methyl thiazolyl tetrazolium （MTT） assays. Cell cycle distrtibutions were detected by flow cytometry. After MG63 cells were treated with 5 mM metformin for 48 h, the expression of cyclin D 1 and protein phosphorylation levels of AMPK, AKT, S6K and S6 were detected by western blotting. Results 1 - 50 mmol/L of metformin concentrations in all the groups showed obvious diffenence in proliferation inhibition of MG63 cells, except two groups of 10 mmol/L and 20 mmol/L of concentrations （P 〈 0.01）. Metformin at 5 mmol/L concentration induced the blocking of G1 phase of MG63 cells （P 〈 0.01）, in inhibiting the expression of cylin D1, P-S6K1 and P-S6, promoting the protein pbospborylation of AMPK, AKT. Conclusion Metformiu inhibites proliferation of MG63 cells by inducing the G1 phase blocking , activating AMPK, AKT protein phosphorylation to inhibite mTOR signa way.