miR-21通过靶向FasL基因对脑胶质瘤细胞生长的影响  

miR-21 influences growth of glioma cells by targeting FasL gene

作　　者：张华[1] 郭文涛[2] 

机构地区：[1]赣南医学院,江西赣州341000 [2]漯河医学高等专科学校,河南漯河462002

出　　处：《中国病理生理杂志》2015年第8期1495-1499,共5页Chinese Journal of Pathophysiology

摘　　要：目的:探讨脑胶质瘤细胞中miR-21对Fas L表达的调控作用以及对细胞生长和凋亡的影响,并研究其分子作用机制。方法:将miR-21模拟物(miR-21 mimics)、miR-21抑制物(miR-21 inhibitor)以及阴性对照(scramble)瞬时转染到U251细胞中,CCK-8法和流式细胞术检测细胞活力和凋亡情况。构建Fas L 3’UTR双萤光素酶报告载体,通过采用双萤光素酶报告实验验证miR-21的靶基因。构建表达载体pc DNA3.1-Fas L,回复实验分析miR-21对细胞凋亡的影响。结果:miR-21过表达可促进U251细胞的活力,抑制细胞凋亡;miR-21表达下调则抑制细胞活力,促进细胞凋亡,和对照组比较差异有统计学意义(P<0.05)。双萤光素酶报告实验和回复实验结果提示miR-21可以通过作用于Fas L的3’UTR区,负向调控其表达,从而抑制细胞的凋亡。结论:miR-21可以通过靶向调控Fas L的表达进而促进U251细胞的生长。AIM： To investigate the regulation of miR-21 on Fas L expression and its effect on the growth and apoptosis in glioma cells,and to evaluate the molecular mechanism. METHODS： Differential expression levels of miR-21 in human glioma U251 cells were achieved by transfecting with miR-21 mimics,miR-21 inhibitor or scramble. The viability and apoptosis of U251 cells were detected by CCK-8 assay and flow cytometry with Annexin V- FITC / PI double staining.The recombination vector pmir GLO-Fas L was constructed. Dual-luciferase reporter experiment was performed to validate the target genes of miR-21. The expression vector pc DNA3. 1-Fas L was also constructed,and the biological activity and regulatory role of miR-21 in U251 cell apoptosis were analyzed by a restore experiment. RESULTS： Exogenous overexpression of miR-21 increased the viability and decreased the apoptosis of U251 cells（P〈0. 05）,while miR-21 inhibitors generated the opposite results（P〈0. 05）. Dual-luciferase reporter assay and restore experiment revealed that miR-21 negatively regulated the expression of Fas L gene which was regarded as the target gene,thus decreasing the apoptosis of U251 cells.CONCLUSION： miR-21 increases the viability of glioma U251 cells,in which Fas L may be one of the target genes.

关 键 词：胶质瘤 微小RNA.21 细胞凋亡 FAS配体 

分 类 号：R730.23[医药卫生—肿瘤]