RNA干涉脑胶质瘤MSP58基因后肿瘤转移相关基因的表达变化  

Expression changes of tumor metastasis related gene after MSP58 gene RNA interference in human brain glioma

作　　者：林伟[1] 罗小楠[2] 章翔[1] 张璟[3] 张健[3] 费舟[1] 

机构地区：[1]第四军医大学西京医院神经外科,陕西西安710032 [2]第四军医大学训练部临床管理处,陕西西安710032 [3]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》2010年第1期4-9,共6页Chinese Journal of Neurosurgical Disease Research

基　　金：国家自然科学基金资助项目(30672371);陕西省科技计划基金资助项目(2008K09-09)

摘　　要：目的采用基因芯片技术,观察RNA干涉法抑制人脑胶质瘤细胞系U251细胞MSP58基因表达后肿瘤转移相关基因的变化,从而了解MSP58在肿瘤转移相关基因通路中的可能定位。方法将特异性的MSP58干涉表达载体pSilencer3.1-MSP58转染人脑胶质瘤细胞系U251,同时构建相应的阴性对照载体pSilencer3.1-NC以及空载体pSilencer3.1-H1neo。运用半定量RT-PCR及Western blot的方法检测稳定转染和未转染干涉载体的胶质瘤细胞MSP58的mRNA和蛋白的表达水平。运用基因芯片技术分析pSilencer3.1-MSP58转染后肿瘤转移相关基因表达的变化。结果成功建立了具有稳定下调MSP58基因表达的U251-S细胞、阴性对照U251-NC细胞以及含有空载体的U251-H1neo细胞。干涉组细胞的MSP58的表达无论在mRNA水平还是在蛋白水平均较阴性对照组及空载体组细胞显著降低。基因芯片技术共筛选肿瘤转移相关基因128个,其中上调2倍以上的共有34个基因。结论干涉MSP58基因表达后,胶质瘤细胞增殖、迁移和侵袭能力的下降与肿瘤转移相关基因表达的变化有关。Objective To investigate the expression changes of tumor metastasis related gene after suppression of MSP58 by RNA interference in the glioma U251 cells using DNA microarray in order to probe the role of MSP58 in tumor metastasis related gene pathway. Methods Human glioma cells （ U251 ） were transfected with the specific siRNA expression vector （pSilencer3. 1-MSP58 ） designed to target MSP58 mRNA. A corresponding site-mutated vector was constructed as negative control （ pSilencer3.1. NC）. The expressions of MSP58 mRNA and its protein among the stable transfected cells and the untransfected ones were detected by semi-quantitative RT-PCR and Western blot, respectively. Results Three stable transfected cell lines： U251-S （with pSileneer3.1-MSPS）, U251-NC （with pSilencer3.1-NC） and U251- H1 neo （with empty vector pSilencer3.1-H1 neo） were established. The expression levels of MSP58 mRNA and protein in U251-S were significantly lower than that in U251-NC, U251-H1 neo. A total of 128 tumor metastasis related genes were examined by DNA microarray. Thirty-four genes were up-regulated by over 2 folds but no one was down-regulated by more than 2 folds. Conclusion The inhibition of human glioma cell proliferation, migration and invasion by MSP58 gene RNA interference may be concerned with the expression changes of tumor metastasis related gene.

关 键 词：基因芯片 RNA干涉 微小球蛋白 胶质瘤 

分 类 号：R739.41[医药卫生—肿瘤]