微小RNA-100下调S100A4表达抑制膀胱癌细胞迁移  

Inhibitory effects of microRNA-100 on migration of human bladder urothelial carcinoma cells through down-regulating S100A4 expression

作　　者：刘辉[1] 贾招辉[1] 王晓彬[1] 史海军[1] 魏曙光[1] 窦中岭[1] 

机构地区：[1]河南科技大学第一附属医院泌尿外科,洛阳471003

出　　处：《中华实验外科杂志》2015年第6期1227-1229,共3页Chinese Journal of Experimental Surgery

基　　金：河南省教育厅科学技术研究重点项目（13A3203X3）

摘　　要：目的 观察在膀胱癌细胞中微小RNA（miRNA,miR）-100对靶基因S100A4的调控及细胞迁移的影响.方法 基于Sanger数据库对miR-100进行靶基因预测,利用慢病毒Lenti-miR-100、Lenti-shS100A4感染膀胱癌细胞EJ、T24,应用实时定量反转录聚合酶链反应（RT-qPCR）、Western blot检测S100A4mRNA和蛋白的表达,并通过Transwell迁移实验检测高表达miR-100及敲低S100A4对膀胱癌细胞EJ、T24迁移能力的影响.结果 RT-qPCR和Western blot表明,与对照细胞比较,高表达miR-100膀胱癌细胞EJ、T24中S100A4表达水平明显受到抑制（P＜0.01）,细胞的迁移数为（130.0±3.2）个,明显低于对照组[（450.0±1.5）个,P＜0.01];S100A4基因沉默可明显抑制膀胱癌细胞EJ、T24迁移能力,为（137.0±5.3）个（P＜0.01）.结论 miRNA-100可通过下调S100A4表达抑制膀胱癌细胞迁移.Objective To explore the effects of microRNA-100 （miRNA,miR-100） on S100A4 and migration in human bladder urothelial carcinoma cells.Methods We established human bladder urothelial carcinomo cell transfectants stably expressing miR-100 and SiS100A4 using lentivirus infection.The expression levels of miR-100 and S100A4 were determined by real-time quantitative reverse transcriptase-polymerase chain reaction （RT-qPCR） and Western blotting.The relationship between S100A4 and migration of EJ cells was evaluated using Transwell assays.Results S100A4 mRNA and protein expression levels were significantly decreased in human bladder urothelial carcinoma cells after transfection with lenti-miR-100 （P ＜ 0.01）.Meanwhile,silence of endogenous S100A4 by lenti-SiS100A4 could reduce the migration.The results of transwell assay showed that the number of migrating cells in high miR-100 expression group was 130.0 ± 3.2,significantly lower than that in control group （450.0 ± 1.5,P ＜ 0.01）.After S100A4 down-regulation,the number of migrating cells was strongly reduced （137.0 ± 5.3,P ＜ 0.01）.Conclusion Stable expression of miR-100 reduced migration abilities via down-regulating S100A4 in human bladder urothelial carcinoma cells.

关 键 词：微小RNA-100 膀胱癌 S100A4 迁移 

分 类 号：R737.14[医药卫生—肿瘤]