miR-17靶向调控MMP-2基因对人脑胶质瘤U87细胞侵袭性的影响  

Role of miR-17 in human glioma U87 cells invasion by regulating MMP-2 expression

作　　者：周青青[1] 张振兴[1] 李晨光[1] 李洋[1] 王洪磊[1] 

机构地区：[1]辽宁医学院附属第一医院神经外科,辽宁锦州121001

出　　处：《军事医学》2015年第11期821-824,共4页Military Medical Sciences

基　　金：辽宁省高等学校杰出青年学者成长计划基金资助项目(LJQ2013088)

摘　　要：目的探讨人脑胶质瘤U87细胞中微小RNA-17(microRNA-17,miR-17)和基质金属蛋白酶(matriv metalloproteinase,MMP)-2基因的表达,阐明miR-17对MMP-2基因的靶向调控作用以及对U87细胞侵袭性的影响。方法培养U87细胞并应用免疫荧光化学技术检测MMP-2基因的表达;采用生物信息学方法对miR-17和MMP-2基因的匹配关系进行预测并应用双荧光素酶报告基因系统鉴定;转染miR-17模拟物后,行实时PCR检测miR-17和MMP-2基因的表达,Western印迹检测MMP-2蛋白的表达,Transwell小室检测癌细胞体外的侵袭性。结果倒置相差显微镜下可见U87细胞分布均匀,多样性明显;细胞免疫荧光表明胞质内有MMP-2蛋白的表达,显示绿色荧光。靶基因预测软件miRanda和Target Scan显示miR-17和MMP-2基因匹配良好,双荧光素酶报告基因系统鉴定发现miR-17可结合MMP-2 mRNA 3'UTR并有效抑制其表达。实时PCR和Western印迹检测结果均表明miR-17的过表达可下调MMP-2基因表达。Transwell实验表明miR-17的过表达可抑制U87细胞的侵袭。结论 miR-17通过靶向作用于MMP-2 mRNA 3'UTR负性调控人脑胶质瘤U87细胞中MMP-2的表达并可抑制其侵袭作用。Objective To identify the role of microRNA-17（miR-17）in human glioma U87 cells invasion which may regulate expression of matriv metalloproteinase（MMP）-2.Methods U87 cells were cultured in vitro,while changes in cellular morphology were observed by phase contrast microscope.The miR-17 which might regulate the expression of MMP-2 was predicted by bioinformatics and identified using dual luciferase report system.Expressions of miR-17 and MMP-2 were determined using real-time PCR and Western blot after transfection of miR-17 mimics.The invasion of U87 cells was detected in vitro by Transwell chamber.Results Expression of MMP-2 was positive by immunofluorescence cytochemistry. Using dual luciferase reporter system,miR-17 could inhibit the expression of MMP-2 by binding to its mRNA 3′UTR. Results of real-time PCR and Western blot showed that over-expression of miR-17 down-regulated expression of MMP-2. The invasion of U87 cells was suppressed by over-expression of miR-17.Conclusion MiR-17 may negatively regulate expression of MMP-2 in human glioma U87 cells and inhibit cell invasion.

关 键 词：基质金属蛋白酶-2 人脑胶质瘤 U87 细胞 MMP-2 

分 类 号：R739.41[医药卫生—肿瘤]