中华神经外科杂志
中華神經外科雜誌
중화신경외과잡지
Chinese Journal of Neurosurgery
2014年
4期
409-413
,共5页
谭富强%张安玲%南阳%王乐%叶敏华%王广秀%贾志凡%康春生%钟跃
譚富彊%張安玲%南暘%王樂%葉敏華%王廣秀%賈誌凡%康春生%鐘躍
담부강%장안령%남양%왕악%협민화%왕엄수%가지범%강춘생%종약
亮氨酰-tRNA合成酶%神经胶质瘤%细胞增殖%细胞迁移%小干扰RNA
亮氨酰-tRNA閤成酶%神經膠質瘤%細胞增殖%細胞遷移%小榦擾RNA
량안선-tRNA합성매%신경효질류%세포증식%세포천이%소간우RNA
Leucyl-tRNA synthetase%Glioma%Cell proliferation%Cell migration%RNA,small interfering
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(LARS siRNA)于U251.PCR、Western blot检测LARS表达;MTT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LARS siRNA后LARS mRNA和蛋白水平明显降低,自转染48 h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.
目的 探究敲低亮氨酰-tRNA閤成酶(LARS)錶達對U251細胞增殖和遷移能力的影響.方法 採用脂質體轉染LARS小榦擾RNA(LARS siRNA)于U251.PCR、Western blot檢測LARS錶達;MTT法、平闆剋隆檢測細胞增殖能力、流式細胞術檢測細胞週期及Transwell法檢測細胞遷移能力.結果 較對照組和無義序列組,轉染LARS siRNA後LARS mRNA和蛋白水平明顯降低,自轉染48 h後細胞增殖率明顯下降(P＜0.05),且剋隆形成率分彆較對照組和無義序列組降低瞭51.69％和50.45％,G0/G1期比例分彆增高瞭38.34％和31.93％,穿膜細胞數分彆減少瞭81.78％和81.45％,差異均有統計學意義(P＜0.05).結論 敲低LARS錶達可明顯抑製U251生長增殖和遷移能力.LARS基因有望成為人腦膠質瘤基因治療的侯選靶點.
목적 탐구고저량안선-tRNA합성매(LARS)표체대U251세포증식화천이능력적영향.방법 채용지질체전염LARS소간우RNA(LARS siRNA)우U251.PCR、Western blot검측LARS표체;MTT법、평판극륭검측세포증식능력、류식세포술검측세포주기급Transwell법검측세포천이능력.결과 교대조조화무의서렬조,전염LARS siRNA후LARS mRNA화단백수평명현강저,자전염48 h후세포증식솔명현하강(P＜0.05),차극륭형성솔분별교대조조화무의서렬조강저료51.69％화50.45％,G0/G1기비례분별증고료38.34％화31.93％,천막세포수분별감소료81.78％화81.45％,차이균유통계학의의(P＜0.05).결론 고저LARS표체가명현억제U251생장증식화천이능력.LARS기인유망성위인뇌효질류기인치료적후선파점.
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 MTT 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 G0/G1phase increasing 38.34％ and 31.93％ (P ＜0.05)and the number of cell passing through transwell membrane was reduced markly by 81.78 ％ and 81.45 ％ (P ＜ 0.05).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.
