敲低亮氨酰-tRNA合成酶抑制人胶质瘤细胞U251增殖和迁移的体外研究  

Implication of leucyl - tRNA synthetase in proliferation and migration of human glioma cell line U251 detected by siRNA targeted knock - down analysis

作　　者：谭富强[1] 张安玲[1] 南阳[1] 王乐[1] 叶敏华[1] 王广秀[1] 贾志凡[1] 康春生[1] 钟跃[1] 

机构地区：[1]天津医科大学总医院神经外科、天津市神经病学研究所神经肿瘤实验室,300052

出　　处：《中华神经外科杂志》2014年第4期409-413,共5页Chinese Journal of Neurosurgery

基　　金：天津市自然科学基金（13JCQNJC10800,13JCZDJC31000）

摘　　要：目的探究敲低亮氨酰-tRNA合成酶（LARS）表达对U251细胞增殖和迁移能力的影响。方法采用脂质体转染LARS小干扰RNA（LARSsiRNA）于U251。PCR、Westernblot检测LARS表达；MTr法、平板克隆检测细胞增殖能力、流式细胞术检测细胞周期及Transwell法检测细胞迁移能力。结果较对照组和无义序列组，转染LARSsiRNA后LARSmRNA和蛋白水平明显降低，自转染48h后细胞增殖率明显下降（P〈0．05），且克隆形成率分别较对照组和无义序列组降低了51．69％和50．45％，G0／G1期比例分别增高了38．34％和31．93％，穿膜细胞数分别减少了81．78％和81．45％，差异均有统计学意义（P〈0．05）。结论敲低LARS表达可明显抑制U251生长增殖和迁移能力。LARS基因有望成为人脑胶质瘤基因治疗的侯选靶点。Objective To investigate the effect of leucyl - tRNA synthetase （LARS） by siRNA knock- down on proliferation and migration of human glioma cell line U251. Methods Artificially synthesized small interfering RNA for LARS gene （LARS siRNA） was transfected into U251 cell by Lipofectamine2000. LARS expression in transfected cells was detected by real -time PCR and western blot. The growth and migration ability of U251 cells were observed by using a serial of experimental technology including MTF assay, plate colony formation assay, flow cytometry and transwell assay. Results LARS siRNA effectively knocked down the LARS expression in U251 cells. Compared with control and scramble groups, the proliferation activity was significantly suppressed （ P 〈 0. 05 ） from second day and plate colony formation efficiency was decreased by 51.69% and 50. 45% （ P 〈 0.05 ） and cell cycle was arrested in Go/G1 phase increasing 38.34% and 31.93 % （ P 〈 0.05 ）and the number of cell passing through transwell membrane was reduced markly by 8 I. 78 % and 81.45 % （ P 〈 O. 05 ）. Conclusions Knock - down LARS by siRNA could inhibit the proliferation activity and migration ability of glioma cells. These preliminary founding suggested that LARS might play an oncogenic role in the proliferation and migration of glioma cell, with serving as a potential target of gene therapy for glioma.

关 键 词：亮氨酰-TRNA合成酶 神经胶质瘤 细胞增殖 细胞迁移 小干扰RNA 

分 类 号：R739.41[医药卫生—肿瘤]