BRG1基因对胶质瘤细胞转移潜能影响及其机制探讨  

Silence of BRG1gene regulates migration and invasion of human glioma cells and its molecular mechanisms

作　　者：时梅林[1,2] 梅鹏金[3] 范月超[3] 李中林[3] 白津 徐凯[2] 郑骏年 

机构地区：[1]江苏省肿瘤生物治疗重点实验室,江苏徐州221002 [2]徐州医学院医学影像学院,江苏徐州221002 [3]徐州医学院附属医院神经外科,江苏徐州221002

出　　处：《中华肿瘤防治杂志》2013年第18期1395-1398,共4页Chinese Journal of Cancer Prevention and Treatment

摘　　要：目的:探讨沉默BRG1基因对胶质瘤细胞迁移和侵袭的影响及分子机制。方法:体外化学合成BRG1siRNA和对照siRNA(si-Ctrl),脂质体介导转染胶质瘤U251和U87细胞,Transwell迁移实验和侵袭实验观察沉默BRG1基因对两种胶质瘤细胞迁移和侵袭能力的影响,明胶酶谱实验检测MMP-2变化,蛋白质印迹法检测TIMP-2和MMP2蛋白表达。结果:Transwell迁移实验中,转染BRG1siRNA组U251细胞穿越Transwell小室的细胞个数为23.13±1.20,与对照组96.38±9.67相比,减少了76%,P=0.000 1;U87穿越细胞个数为37.33±2.31,对照组为233.31±19.37,减少了84%,P<0.001。侵袭实验中,转染BRG1siRNA组U251细胞穿越Transwell小室的细胞个数为26.27±1.05,与对照组97.30±9.35比较,减少了73%,P=0.000 1;U87穿越细胞个数为32.83±2.42,对照组为234.50±15.67,减少了86%,P<0.001。与对照组相比,蛋白质印迹法显示,BRG1沉默后TIMP-2表达升高,MMP-2表达下降;明胶酶谱实验显示,BRG1沉默后MMP-2酶活性降低。结论:BRG1基因沉默通过上调TIMP-2抑制MMP-2的表达,破坏TIMP-2/MMP-2平衡,最终抑制胶质瘤细胞迁移和侵袭能力。OBJECTIVE：To investigate the effect of BRG1 gene in glioma cell migration and invasion and its molecu- lar mechanism. METHODS：Using chemically synthesized small interfering RNA,we transfected BRG1 siRNA into human glioma cell lines U251 and U87 by siLentFect Lipid Reagent. We studied the role of BRG1 in glioma cell migration and in- vasion by cell migration assay and matrigel invasion assay. We performed western blot to detect TIMP-2 and MMP-2 pro- tein expression. We also detected MMP-2 enzyme activity by gelatin zymography. RESULTS：Compared to the control siR- NA treatment,knock down of BRG1 decreased the number of U251 and U87 cells that passed the transwell membrane re- spectively by 76%（96.38±9.67 vs 23.13±1.20,P=0. 000 1） and 84% （233.31±19.37 vs 37.33±2.31,P〈0. 001）, which were detected by transwell migration assay. Matrigel invasion assay showed that knock down of BRG1 decreased the number of U251 and U87cells that passed the transwell membrane respectively by 73% （97.30± 9.35 vs 26.27±1.05, P=0. 000 1） and 86% （234.50±15.67 vs 32.83±2.42,P（0. 001） ,which were compared to the control group. Western blotting showed that knock down of BRG1 increased TIMP-2 expression and decreased MMP-2 expression,and gelatin zy- mography showed that knock down of BRG1 decreased MMP-2 enzyme activity. CONCLUSIONS：Our data indicated that silence of BRG1 in glioma cells inhibited cell migration and invasion abilities. Up-regulation of TIMP-2 expression and down-regulation of MMP-2 expression greatly contributed to the reduced cell invasion and migration abilities.

关 键 词：脑肿瘤 BRG1 侵袭 迁移 MMP-2 TIMP-2 

分 类 号：R739.4[医药卫生—肿瘤]