MSP58基因小干扰RNA对胶质瘤细胞系U251增殖的影响  

Effect of small interfering RNA targeting MSP58 gene on proliferation of human glioma cell line U251

作　　者：王江[1] 林伟[1] 章翔[1] 张璟[2] 张健[2] 吴琳[2] 刘新平[2] 药立波[2] 

机构地区：[1]第四军医大学西京脑科医院神经外科 [2]第四军医大学基础部生物化学与分子生物学教研室,陕西西安710032

出　　处：《中华神经外科疾病研究杂志》2009年第1期10-14,共5页Chinese Journal of Neurosurgical Disease Research

基　　金：国家自然科学基金资助项目(39970752)

摘　　要：目的观察核微球蛋白58(MSP58)基因特异性siRNA(small interfering RNA)对神经胶质瘤U251细胞的体外增殖的影响。方法体外化学合成针对MSP58基因的siRNA,脂质体法转染神经胶质瘤细胞系U251,荧光显微镜观察转染效率,逆转录酶-多聚酶链反应(RT-PCR)和Western blot分别检测MSP58基因的mRNA和蛋白表达水平,甲基噻唑基四唑法(MTT法)绘制细胞生长曲线,流式细胞仪(FCM)分析细胞周期。结果MSP58基因的特异性siRNA转染U251细胞后,MSP58的mRNA和蛋白表达水平明显降低,细胞增殖受到明显抑制;流式细胞仪检测显示转染后的U251细胞G2/M期细胞明显减少(P<0.01),S期细胞略有减少,G1期细胞明显增加(P<0.01),G1期发生明显阻滞。结论MSP58基因的特异性siRNA可明显抑制神经胶质瘤细胞U251的增殖,并引起G1期细胞阻滞。Objective To observe the influence of specific small interfering RNA（siRNA）targeting 58-kDa microspherule protein（MSP58）gene on the proliferation of glioma cell line U251 in vitro.Methods The siRNA targeting MSP58 gene and negative control siRNA were synthesized chemically in vitro,which was transfected transiently respectively into glioma cell line U251 by Lipofectamine TM2000.The transfection efficiency of siRNA targeting MSP58 gene was observed by fluorescence microscope.Reverse transcriptase-polymerase chain reaction（RT-PCR）and Western blot were used to investigate the expression of mRNA and protein of MSP58.The cell growth curve was drawn by methyl thiazolyl tetrazolium assay（MTT）.Flow cytometry was adopted to analyze cell cycle.Results After transfecting the specific siRNA targeting MSP58 into U251 cells,the mRNA and protein level of MSP58 declined markedly,and the cell proliferation was significantly inhibited.G2/M phase cells decreased notably（P〈0.01）,S phase cells reduced unclearly and G1 phase cells increased apparently（P〈0.01）.Conclusion The specific siRNA targeting MSP58 can significantly inhibit the proliferation of U251 cells and induce cell cycle arrest at G1 phase.

关 键 词：MSP58 SIRNA 神经胶质瘤 

分 类 号：R739.41[医药卫生—肿瘤]