miR-106a抑制胶质瘤细胞增殖并诱导凋亡

Overexpression of miR-106a inhibits proliferation and induces apoptosis of glioma cells

目的胶质瘤是最常见的原发性中枢神经系统肿瘤,具有较高的病死率。现有的研究表明miR-106a在多种肿瘤中表达上调,并发挥着致癌性作用,但miR-106a在胶质瘤中的表达和作用却并不清楚。方法不同级别胶质瘤组织和胶质瘤细胞系(T98G,SHG44,U87,U251,U373)内的miR-106a水平通过实时定量聚合酶链式反应(qRT-PCR)测定。转染miRNA寡聚核苷酸后的U87和U251细胞的增殖能力和细胞周期分别采用四甲基偶氮唑盐(MTT)比色法和流式细胞仪测定,并且通过膜联蛋白/碘化丙啶(annexin V/PI)双染法测定了miR-106a对胶质瘤细胞的凋亡影响。结果miR-106a在胶质瘤组织和胶质瘤细胞系内表达水平相对正常脑组织显著下降,并且miR-106a表达水平与胶质瘤病理级别负相关。胶质瘤细胞转染miR-106a后,可检测到过表达的miR-106a可显著抑制胶质瘤细胞的增殖能力,阻滞细胞周期于G0/G1期,同时诱导胶质瘤细胞凋亡增加。结论 miR-106a可阻滞胶质瘤细胞细胞周期、抑制增殖和诱导凋亡。

Objective To investigate the effect of miR-106a on proliferation and apoptosis of glioma cells.Methods MiR-106a levels in glioma tissues of different grades and glioma cell lines（T98G,SHG44,U87,U251,U373） were measured with quantitative real-time polymerase chain reaction（qRT-PCR）.miRNA oligonucleotides were transfected into U87 and U251 using lipofectamine 2000 reagen at the time of 70%~90% confluence.The in vitro growth ability of transfected cells was determined by dimethyl thiazolyl diphenyl tetrazolium（MTT） assay,and the cell-cycle distribution was analyzed using flow cytometry（FCM）.After MTT and cell-cycle distribution analysis,the effect of miR-106a on apoptosis was tested by annexin V and propidium iodide（PI） double staining.Results The expression of miR-106a in both glioma specimens and cell lines was significantly lower than that in NATs.Moreover,miR-106a expression was lower in grade III and grade IV glioma compared to grade I and grade II tumors.After miR-106a mimic,inhibitor or scrambled control was transfected in human glioma cells.The results showed that overexpression of miR-106a could suppress the glioma cells proliferation and increase the apoptosis.Conclusion miR-106a induces apoptosis and inhibits proliferation of glioma cells and it may play a potential role in glioma therapy.