二甲双胍对AGS胃癌细胞生长侵袭的抑制作用

Metformin inhibits cell proliferation and migration in gastric cancer cell line AGS

目的:研究应用安全性较好的二甲双胍对胃癌细胞的抑制作用,初步探讨可能机制.方法:以二甲双胍联合或不联合5-氟尿嘧啶干预AGS细胞作为实验组,无药物组作对照.干预24、48、72h应用MTT检测细胞增殖,干预48h流式细胞术检测凋亡、线粒体膜电位,72h通过细胞划痕实验反映迁移能力改变,药物作用24h抽提细胞RNA以RT-PCR检测相关基因转录改变.结果:AGS细胞相对活力明显降低(24、48、72h:F=99.32,127.30,235.72,均P<0.01),并具有浓度-时间依赖性,联合应用5-氟尿嘧啶显示协同作用(24h:t=2.97,P<0.05;48、72h:t=4.61,6.02,均P<0.01).线粒体膜电位下降(t=12.43,P<0.01),凋亡增加(t=8.32,P<0.01),平均迁移速率明显减慢(12、24、48h:t=9.13,13.77,14.21,均P<0.01),cyclinD1、Bcl-2、MMP-2、MMP-9mRNA均减少,Bax增加.结论:二甲双胍能够抑制AGS胃癌细胞增殖、迁移,促进凋亡,与5-氟尿嘧啶联合应用具有协同抑制作用.

AIM：To investigate the effects of metformin,a widely used antihyperglycemic agent,on cell proliferation,apoptosis and migration in human gastric cancer cell line AGS,and to examine whether there are synergistic effects between metformin and 5-？ uorouracil （5-FU）.METHODS：AGS cells were treated with metformin alone or in combination with 5-FU.Methyl thiazolyl tetrazolium （MTT） assay was used to detect cell relative viability at 24,48 and 72 h after treatment.Apoptosis and mitochondrial membrane potential were examined by flow cytometry （FCM） at 48 h.Cell migration at 72 h was determined by scratch assay.The expression of cyclin D1,Bcl-2,MMP-2,MMP-9 and Bax mRNAs in AGS cells at 24 h was detected by RTPCR.RESULTS：Metformin significantly inhibited cell proliferation in a dose-and time-dependent manner （24,48 and 72 h：F=99.32,127.30 and 235.72,respectively;all P 〈0.01）.After metformin treatment,mitochondrial membrane potential decreased （t=12.43,P 〈0.01）,apoptosis rate increased （t=8.32,P 〈0.01）,and average migration velocity was reduced （12,24 and 48 h：t=9.13,13.77 and 14.21,respectively;all P 〈0.01） in AGS cells.The expression of cyclin D1,Bcl-2,MMP-2 and MMP-9 mRNAs was down-regulated,while that of Bax mRNA was up-regulated after metformin treatment.Metformin treatment enhanced 5-FU-mediated cell growth inhibition （24 h：t=2.97,P 〈0.05;48 h,t=4.61,P 〈0.01;72 h：t=6.02,P 〈0.01）.CONCLUSION：Metformin can inhibit cell proliferation and migration and promote apoptosis in human gastric cancer cell line AGS.Metformin and 5-FU have a synergistic anti-proliferation effect in AGS cells.