miR-370-3p对胶质母细胞瘤U87-MG细胞株增殖能力的影响

Effects of Mirco RNA-370-3p on proliferation of glioma cell line U87-MG and its potential mechanism

目的探讨微小RNA-370-3p(miR-370-3p)对人脑胶质瘤细胞系U87-MG增殖能力的影响及其机制。方法将常规培养的U87-MG细胞分为空白组、无义序列转染组和模拟物转染组,后两组分别转染miRNA无义序列及miR-370-3p模拟物,qRT-PCR法检测其转染效率,免疫印迹法检测转染细胞叉头框蛋白M1(FoxM1)的表达;采用Ed U法评估细胞的增殖能力,采用平板克隆形成实验检测细胞的增殖能力。结果与空白组和无义序列转染组比较,模拟物转染组细胞miR-370-3p表达显著增加(P<0.05),而FoxM1的蛋白表达显著减少(P<0.05);而且模拟物转染组U87-MG细胞增殖能力及克隆形成率均明显减少(P<0.05)。结论 miR-370-3p能够抑制U87-MG细胞的增殖能力,可能与减少FoxM1的表达有关。

Objective To explore the effect of Mirco RNA（mi RNA）-370-3p on the proliferation of U87-MG glioblastoma cells and its mechanism.Methods The cultured U87-MG cells were divided into blank group, mimics transfection group（transfected with Has-mi RNA-370-3p mimics） and negative control group（transfected with mi RNA-ctrl）. Transfections of Has-mi RNA-37-3p mimics were mediated by lipofectamine 2000 U87-MG cells. The transfection efficiency were detected by quantative real-time PCR. The expression of Forkhead M1（FoxM1） protein were investigated by Western blotting 48 h after the transfection. The cell proliferation ability was evaluated by 5-ethynyl-2＇-deoxyuridine（EdU） incorporation and colony formation assaies 72 h after the transfection.Results The expression of mi RNA-370-3p increased and FoxM1 protein expression decreased significantly in U87-MG cells in the mimics transfection group compared with those in the blank group and negative control group. EdU assay showed that the short-term ability of the U87-MG cells proliferation was significantly lower in the mimics transfected group than those in the other two groups（P〈0.05）. The colony formation assay showed that the long-term ability of the U87-MG cells proliferation was significantly lower in themimics transfection group than those in the other two groups（P〈0.05）.Conclusion It is suggested that upregulation of mi RNA-370-3p expression can inhibit proliferation of U87-MG cells, probably by downregulating expression of FoxM1.