miR-218对胶质母细胞瘤细胞成熟分化的影响  

Stimulatory Effects of miR-218 on Glioblastoma Cell Differentiation

作　　者：孔妍玲[1] 王虔[1] 于士柱[1] 孙翠云[1] 李艳艳[1] 张景敏[1] 徐金玲[1] 王影[1] 安同岭[1] 温艳军[1] 王菲[1] 魏常娟[1] 孙静[1] 刘静[1] 

机构地区：[1]天津医科大学总医院天津市神经病学研究所,天津市神经损伤变异与再生重点实验室,教育部中枢神经创伤修复与再生重点实验室,天津市300052

出　　处：《中国肿瘤临床》2012年第9期532-535,共4页Chinese Journal of Clinical Oncology

基　　金：国家973计划分项目(编号:2010CB529405);天津市应用基础及前沿技术研究计划重点项目(编号:10JCZDJCl9400);天津市科技创新体系及条件平台建设计划重大项目(编号:10SYSYJC28800)资助~~

摘　　要：目的:观察miR-218对胶质母细胞瘤细胞系成熟分化的影响。方法:通过质粒转染和G418筛选建立稳定过表达miR-218的SNB-19亚细胞系及对照亚细胞系。利用qRT-PCR及Western blot检测稳定细胞系中miR-218、CD133及Nestin表达水平。用GFAP免疫细胞化学染色评价其成熟分化水平,通过免疫荧光观察转染miR-218表达质粒后CD133及Nestin阳性细胞的分布情况。结果:过表达miR-218的亚细胞系中miR-218、CD133及Nestin表达水平分别是对照细胞的26.23倍、17.9%及54.2%,过表达miR-218细胞的GFAP阳性标记指数为(96.0±3.81)%显著高于对照细胞(49.6±5.13)%(t=16.24,P<0.0)1)。免疫荧光显示CD133及Nestin的下调特异性发生于成功转染miR-218表达质粒的细胞。结论:miR-218可通过促进胶质瘤干细胞的成熟分化显著下调CD133及Nestin的表达水平,该作用可能是miR-218抑制胶质瘤细胞增殖的重要机制之一。Objective： To investigate the stimulatory effects of miR-218 on the differentiation of glioblastoma cells. Methods： The SNB-19 glioblastoma subcell line overexpressing miR-218 （ SNB19-miR218 ） and the control subcell line overexpressing a non- sense scrambled sequence （ SNB19-scr ） were established via plasmid transfection and G418 screening. The expression levels of miR-218, CD133, and nestin were detected via quantitative reverse transcription polymerase chain reaction and western blot analysis, whereas the differentiation status was assessed via glial fibrillary acidic protein （ GFAP ） immunocytochemistry. Using low-copy plas- mid transfected cells, the distribution of CD133- and nestin-positive cells was detected via immunofluorescence against an enhanced green fluorescent protein （ EGFP ） reporter. Results： The expression levels of miR-218, CD 133, and nestin in the cells transfected with SNBI9-miR218 were 26.23-fold, 17.9%, and 54.2% of those of SNB19-scr. The GFAP labeling index of SNB19-miR218 （ 96.0%± 3.81% ） was significantly higher than that of SNB19-scr （ 49.6% ±5.13%, t = 16.24, P 〈 0.0001 ）. Immunofluorescence showed that the constituent ratios of the CD133- and nestin-positive cells in the EGFP （ ＋ ） and （ - ） subpopulations were significantly uneven （ X 2 = 586.38 and 658.53, respectively, P 〈 0.0001 ）, confirming that the CD133 and nestin were downregulated specifically in the tumor cells successfully transfected with the miR-218 expression plasmid. Conclusion： miR-218 promotes glioma stem cell differentiation, and therefore downregulates CD133 and nestin expression. This effect may be an important mechanism for inhibiting the proliferation of glioma cells.

关 键 词：胶质瘤 微小RNA 胶质瘤干细胞 分化 

分 类 号：R739.4[医药卫生—肿瘤]