二甲双胍对神经胶质瘤生长的抑制作用及其机制研究

Anti- neoplastic activity of metformin in glioma and the mechanisms

目的:探讨二甲双胍对神经胶质瘤细胞生物学活性的影响及其作用机制。方法:以0、5、10、20mmol/L四种浓度的二甲双胍分别对人胶质细胞瘤细胞系U251和T98G进行体外干预,MTT法检测其对细胞增殖的影响;Transwell法检测其对细胞侵袭性的影响;Annexin V/双染法检测其对细胞凋亡的影响;Western blotting检测AMP活化蛋白(AMP-activated protein kinase,AMPK)信号通路相关蛋白的表达水平。结果:二甲双胍可显著抑制神经胶质瘤细胞的增殖,促进细胞的凋亡,而对细胞的侵袭影响不大;Western blot结果表明二甲双胍可增强细胞内AMPK、p38 MAPK的磷酸化蛋白表达水平,同时特异性的p38 MAPK通路阻断剂可抑制二甲双胍的促细胞凋亡作用。结论:二甲双胍可能通过激活神经胶质瘤细胞内的AMPK信号传导通路促进肿瘤细胞的凋亡,以期对临床预防及治疗神经胶质瘤提供理论依据。

Objective：To explore the effects of metformin on biological behavior in human glioma and the mechanisms.Methods：Human glioma U251 and T98G cells were treated with metformin（0,5,10,20mmol/L） in vitro.Cell proliferation was measured by MTT colorimetry,and the invasion ability was determined by Transwell invasion assay.Cell apoptosis was analyzed using flow cytometry by double staining with Annexin Ⅴ and propidium iodide.The expression levels of AMP-activated protein kinase （AMPK）-related proteins were detected by western blotting.Results：Metformin markedly inhibited the proliferation and promoted the apoptosis compared with control group（P ＜0.05）,while,it has no significant effect on cell invasion.The results of Western blot demonstrated that metformin could significantly increase the expression levels of phospho-AMPK and phospho-p38 MAPK proteins in U251 cells,and the specific blocker of p38 MAPK signaling pathway could reverse the pro-apoptosis of metformin in glioma cells.Conclusion：Metformin may promote cell apoptosis through activating AMPK signaling pathway in glioma cells,which provide the theoretical basis for clinical prevention and treatment of glioma.