二甲双胍对低级别非侵袭性膀胱肿瘤细胞增殖的影响

Effects of Metformin on the Proliferation of Noninvasive Low-grade Bladder Cancer Cells

目的:初步探讨降糖药物二甲双胍对膀胱肿瘤细胞253J的作用及其相关作用机制。方法:采用Cell Counting Kit-8(CCK-8)试剂盒分析二甲双胍对膀胱肿瘤细胞增殖的影响。应用流式细胞仪检测二甲双胍对细胞周期及凋亡的影响。并通过免疫印迹方法检测相关蛋白确定可能参与其中的信号分子。结果:在各时间点(24小时,48小时,72小时)二甲双胍处理组与对照组相比膀胱肿瘤细胞的增殖受到明显抑制(P<0.01或P<0.05);与对照组比较,二甲双胍处理组G0/G1期细胞比例上升,S期细胞比例下降(P<0.01或P<0.05);免疫蛋白印迹发现,二甲双胍处理组中的磷酸化AMP激活的蛋白激酶(AMP-activated protein kinase,AMPK)表达升高,同时细胞周期蛋白D1(cyclin D1)的表达下降(P<0.01或P<0.05)。结论:体外实验中二甲双胍能够明显抑制膀胱肿瘤细胞的增殖,通过下调cyclin D1的表达诱导细胞周期停滞于G0/G1期。这些结果表明二甲双胍可能成为治疗膀胱癌的潜在药物。

Objective： To evaluate the effects of metformin on human bladder cancer cell line 253J and its underlying mechanisms. ＇Method.s： Cell Counting Kit-8 （CCK-8） was used to investigate the effects of metformin on 253J cells growth. Flow cytometry was used to evaluate the cell cycle changes after metformin treatment. The possible signaling molecules involved in this process were determined by immunoblot analysis of various proteins. Results： In 24 h, 48 h, and 72 h, the proliferation of bladder cancer cells was significantly inhibited compared with that in control group （P 〈0.01 or P 〈0.05）; Compared with control group, a significant increase was observed in the percentage of cells in the G0/G1 phases. In parallel, there was a reduction in the percentage of cells in the S and G2/M phases （P 〈0.01 or P 〈0.05）. Western blot analysis found that the expression ofphosphorylated AMP-activated protein kinase （AMP-activated protein kinase, AMPK） increased, while the expression of cyclin D 1 decreased in metforrnin-treated group.（P 〈0.01 or P 〈0.05）. Conclusion： Metformin can induce G0/GI cell cycle arrest and inhibit 253J cells proliferation in vitro, suggesting that metforrnin may be a potential therapeutic drug for the treatment of human bladder cancer.