miR-200a通过靶向NCAM1基因影响胶质瘤细胞的迁移及侵袭  

The miR-200a in uences the migration and invasion abilities of glioma cells by targeting NCAM1 gene

作　　者：刘锋[1,2] 王占祥[2] 黄才权[1,2] 沈上杭[2] 徐昊[2] 张绍林[2] 

机构地区：[1]福建医科大学第一临床医学院,福建福州350005 [2]厦门大学附属第一医院神经外科,福建厦门361003

出　　处：《肿瘤》2013年第5期398-403,共6页Tumor

摘　　要：目的:研究微小RNA(microRNA,miRNA)-200a在人神经胶质瘤U87细胞迁移和侵袭中的作用及其可能的机制。方法:利用LipofectAMINETM 2000瞬时转染has-miR-200a模拟物(mimic)、has-miR-200a抑制物(inhibitor)及阴性对照(hsa-miR-negative control),分别上调和下调U87细胞中miR-200a的表达;采用划痕实验和Transwell小室法检测miR-200a对U87细胞迁移及侵袭能力的影响。利用在线miRNA靶基因预测软件预测miR-200a的靶基因,并采用荧光素酶报告系统及蛋白质印迹法对预测的潜在靶基因进行验证。结果:外源性过表达miR-200a可促进U87细胞迁移和侵袭能力,而下调miR-200a的表达则可抑制U87细胞的迁移和侵袭,与对照组比较差异均有统计学意义(P<0.05)。荧光素酶报告系统和蛋白质印迹法检测结果均提示,神经细胞黏附分子(neuralcelladhesionmolecule,NCAM1)基因的蛋白表达受miR-200a的负调控,是其调控的靶基因。结论:miR-200a可以通过靶向调控NCAM1的表达进而促进U87细胞的迁移和侵袭。Objective： To investigate the effects of miR（microRNA）-2OOa on migration and invasion abilities of glioma cells, and to explore its possible mechanism. Methods： Differential expression levels of miR-200a in glioma U87 cells were achieved by transfecting with hsa-miR-2OOa mimic, hsa-miR- 200a inhibitor or hsa-miR-negative control by LipofectAMINETM 2000. The migration and invasion abilities of U87 cells were detected by wound-healing assay and Transwell invasion assay, respectively. Bioinformatics software was used to predict downstream target genes of miR-2OOa and their binding sites. The potential target genes were verified by Luciferase Reporter Assay and Western blotting. Results： Exogenous overexpression of miR-200a could promote migration and invasion abilities of U87 cells （P 〈 0.05）, while miR-200a inhibitors could generate the opposite results （P 〈 0.05）. Luciferase Reporter Assay and Western blotting revealed that hsa-miR-200a negatively regulated the protein expression of NCAM1 （neural cell adhesion molecule 1） gene which was regarded as the target gene. Conclusion： The miR-2OOa can promote the migration and invasion abilities of glioma U87 cells, in which NCAM1 may be one of the target genes.

关 键 词：神经胶质瘤 微RNAS 细胞迁移分析 神经细胞黏附分子类 

分 类 号：R739.41[医药卫生—肿瘤]