miR-9靶向调控跨膜蛋白16A基因对胶质瘤T98G细胞增殖和侵袭的影响  

Effects of miR-9 targeting to TMEM16A on proliferation and invasion of glioma T98G cells

作　　者：李鑫[1] 秦至臻[1] 牛建星[1] 王建祯[1] 

机构地区：[1]北京武警总医院神经肿瘤外科,北京100039

出　　处：《解放军医学院学报》2016年第8期884-887,915,共5页Academic Journal of Chinese PLA Medical School

摘　　要：目的探讨mi R-9和跨膜蛋白16A(transmembrane protein 16,TMEM16A)基因在胶质瘤T98G细胞中的表达,阐明mi R-9对TMEM16A基因的靶向作用及其对T98G细胞增殖和侵袭的影响。方法运用生物信息学方法对mi R-9和TMEM16A基因的靶向配对关系进行预测,采用荧光素酶报告系统鉴定;脂质体2000转染mi R-9模拟物及干扰RNA后,Real-time PCR检测mi R-9与TMEM16A m RNA在癌细胞中的表达,Western blot检测TMEM16A蛋白在癌细胞中的表达,平板克隆形成实验检测癌细胞的增殖情况以及Transwell小室检测癌细胞体外的侵袭性。结果生物信息学软件Target Scan和mi Randa显示mi R-9与TMEM16A基因靶向配对良好,荧光素酶报告系统鉴定发现mi R-9能够抑制TMEM16A m RNA表达。Real-time PCR和Western blot检测结果均表明过表达mi R-9能够降低TMEM16A m RNA和蛋白的表达。平板克隆形成实验和Transwell实验发现过表达mi R-9能够分别抑制T98G细胞的增殖和侵袭。结论 mi R-9通过负性调控胶质瘤T98G细胞中TMEM16A基因的表达抑制癌细胞的增殖和侵袭。Objective To identify expressions of miR-9 and TMEM16 A in glioma T98 G cells, explore the role of miR-9 on cell proliferation and invasion of glioma T98 G cells and TMEM16 A expression. Methods miR-9 which might regulate TMEM16 A expression was predicted by bioinformatics and identified with luciferase assay. After transfection of miR-9 mimics into cells, the expressions of miR-9 and TMEM16 A were determined by real-time PCR and western blot. The proliferation and invasion of T98 G cells were respectively detected in vitro using the colonly formation assay experiment and transwell chamber.Results mi Randa and Target Scan showed that miR-9 was well complementary with TMEM16 A gene. miR-9 could inhibit TMEM16 A m RNA expression shown by luciferase assay. Results of real-time PCR and western blot showed that overexpression of miR-9 could down-regulate the expressions of TMEM16 A m RNA and protein. The proliferation and invasion of T98 G cells were suppressed after transfection of miR-9 mimics. Conclusion miR-9 may negatively regulate TMEM16 A expression in glioma T98 G cells and inhibit cell proliferation and invasion.

关 键 词：微小RNA 跨膜蛋白16A 胶质瘤T98G细胞 

分 类 号：R739.41[医药卫生—肿瘤]