神经胶质瘤干细胞向内皮细胞分化的差异基因表达谱的分析  

Analysis of gene expression profiles in the differentiation process of glioma stem cells to endothelial cells

作　　者：张晋[1] 金贵善[1] 米蕊芳[1] 周益强[1] 房绍伟[2] 朱树干[3] 刘福生[1] 

机构地区：[1]首都医科大学北京市神经外科研究所、首都医科大学附属北京天坛医院神经外科,100050 [2]潍坊医学院 [3]山东大学齐鲁医院神经外科

出　　处：《中华神经外科杂志》2015年第3期299-303,共5页Chinese Journal of Neurosurgery

基　　金：国家自然科学基金（81372354,81302186）;北京市自然科学基金（7132034）;生物医用材料北京实验室资助（京教函【2013】133号）;北京市卫生系统高层次卫生技术人才培养计划（2013-2-018）;山东省自然科学基金（ZR2011HM024）

摘　　要：目的 检测并分析胶质瘤干细胞（GSCs）低氧诱导分化为内皮细胞过程的差异基因表达谱的关键通路和基因,为恶性胶质瘤的治疗寻找新的靶基因.方法 从U87胶质瘤细胞株中提取GSCs细胞并进行免疫荧光鉴定;将GSCs在水凝胶上低氧诱导分化,并对分化后的内皮细胞进行免疫荧光鉴定;收集并提取细胞总RNA,用基因芯片检测分化前、后的细胞差异表达基因,进而分析差异基因的显著靶向性功能和差异基因参与的信号转导通路,从而筛选出关键调控信号通路和基因.结果 提取的GSCs表达CD133、Nestin、SOX2等干细胞标志物;低氧诱导5d后细胞表达CD31、CD34两种内皮细胞标志物;差异基因表达谱存在481个上调基因及245个下调基因（差异倍数＞2）,筛选出转化生长因子β（TGF-β）信号通路、脂肪酸合酶（FASN）、脯氨酰-4-羟化酶亚基α1（P4HA1）等多个有意义的通路和基因（均P ＜0.01）.结论 U87细胞中提取的GSCs可经体外低氧诱导分化为内皮细胞,TGF-β信号通路、FASN及P4HA1可能成为胶质母细胞瘤具有前景的治疗靶点.Objective To detect the different gene expression profiles in the process of glioma stem cells＆#39; （GSCs） differentiation to endothelial cells（ECs） under hypoxia and to screen the key pathways and genes for offering a new therapeutic target for the treatment of glioblastoma.Methods GSCs were extracted from U87 glioma cell line and were identified with the stem cell markers SOX2 、CD133 and Nestin by immunofluorescence staining.The differentiated cells incubated in the hydrogel under hypoxia were identified with the EC markers CD31 and CD34 by immunofluorescence staining.Total RNA extracted from the GSCs and the differentiated cells were used for gene chip to screen expressed genes.The significant gene function analysis （GO-Analysis） and signal transduction pathway analysis （Pathway-Analysis） were carried out to screen the key signaling pathways and genes.Results SOX2、CD133 and Nestin were observed in the GSCs extracted from U87 glioma cell line.CD31 and CD34 were positive in the differentiated cells.Gene chip analysis displayed that 481 genes were up-regulated,while 245 genes were down-regulated.Significant gene function and signal transduction pathways were analyzed and numerous significant signaling pathways and genes were screened such as TGF-β signaling pathway （P 〈 0.01）,FASN （fatty acid synthase）（P 〈0.01） and P4HA1 （prolyl4 hydroxylase subunit alpha-1） （P 〈 0.01）.Conclusions GSCs extracted from U87 glioma cell line could differentiate into ECs under hypoxia in vitro.The significant signaling pathways and genes such as TGF-β signaling pathway,FASN and P4HA1 might be the promising therapeutic targets for the treatment of glioblastoma.

关 键 词：神经胶质瘤 干细胞研究 内皮细胞 细胞低氧 基因芯片 

分 类 号：R739.4[医药卫生—肿瘤]