胶质瘤干细胞中Nanog基因启动子区甲基化检测及意义  

Detection and significance of Nanog promoter methylation in glioma stem cells

作　　者：汪炎[1,2] 牛朝诗[1,2,3] 李仲颖[1,2] 杨洋[1,2] 贺虎[1,2,3] 李冬雪[1,2,3] 

机构地区：[1]安徽医科大学附属省立医院神经外科 [2]脑功能与脑疾病安徽省重点实验室 [3]安徽省脑立体定向神经外科研究所,安徽合肥230001

出　　处：《中华神经外科疾病研究杂志》2014年第5期389-393,共5页Chinese Journal of Neurosurgical Disease Research

基　　金：国家自然科学基金资助项目(81172407);安徽省重点实验室绩效考核基金资助项目(1306c083028)

摘　　要：目的检测胶质瘤干细胞中Nanog基因启动子区甲基化表达,并探讨其Nanog基因表达的关系。方法使用无血清悬浮培养法获得胶质瘤干细胞并鉴定;采用甲基化特异性多聚酶链反应(MSP)分别检测胶质瘤干细胞和胶质瘤细胞系U87中Nanog基因启动子区甲基化状态,实时定量多酶链反应(RT-PCR)检测相应细胞中Nanog表达情况。结果由U87胶质瘤细胞系成功获得胶质瘤干细胞(GSCs);MSP检测胶质瘤干细胞和U87细胞系Nanog启动子区皆呈非甲基化状态,且胶质瘤干细胞中Nanog非甲基化程度高于U87细胞系(t=6.988,P=0.000)。实时定量PCR结果显示胶质瘤干细胞中Nanog mRNA相对表达量高于U87细胞系。结论胶质瘤干细胞中Nanog基因启动子区呈非甲基化状态,可能与Nanog基因表达有关,且可能促进Nanog的转录并维持着胶质瘤干细胞的恶性生物学活性。Objective The methylation of Nanog gene promoter in glioma stem cells（ GSCs） and its association with Nanog gene expression are studied. Methods GSCs were isolated from glioma cell line U87 and cultured in simplified serum-free neural stem cell medium by nanosphere suspension culture method;methylation specific polymerase chain reaction（ MSP） and real-time polymerase chain reaction（ RT-PCR）were conducted to detect the methylation of Nanog gene promoter and its mRNA expression in glioma stem cells and glioma cell lines U87. Results GSCs were isolated,cultured and purified from glioma cell line U87;Nanog gene promoter was hypermethylated in glioma stem cells and U87 cell line. Unmethylation of Nanog gene promoter was higher in glioma stem cells compared to U87 cell line. Nanog mRNA expression in glioma stem cells was higher than that of U87 cell line. Conclusion Unmethylation of Nanog gene promoter regions is a possible mechanism that up-regulates its protein expression and maintains the biological activity of malignancy in glioma stem cells.

关 键 词：胶质瘤 NANOG基因 DNA甲基化 肿瘤干细胞 

分 类 号：R739.41[医药卫生—肿瘤]