生理氧环境下CD133分子在脑胶质瘤干细胞的表达  

Expression of CD133 in Glioma Stem Cells under Hypoxia

作　　者：曾令成[1] 万锋[1] 叶飞[1] 韩林[1] 郭东生[1] 雷霆[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院神经外科,武汉430030

出　　处：《华中科技大学学报（医学版）》2012年第4期423-426,430,共5页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：国家自然科学基金资助项目(No.30801177;No.30901749)

摘　　要：目的通过对生理氧环境下CD133表达的研究,判断脑胶质瘤中CD133mRNA、CD133蛋白以及糖基化CD133即AC133表达与胶质瘤干细胞表型的相关性。方法将CD133阳性胶质瘤干细胞由含20%O2的大气氧环境转换至含1.5%O2的生理氧环境下培养,持续3d。实时荧光定量PCR检测CD133mRNA水平的表达,Western blot检测CD133蛋白水平的表达,流式细胞学检测细胞表面及细胞内AC133的表达,ELISA检测培养液上清中AC133的表达。结果生理氧环境下CD133阳性胶质瘤干细胞NCH421k,NCH441及NCH440细胞表面AC133的表达相对于大气氧环境均显著上调(均P<0.05),其蛋白荧光表达强度分别由(2 381.07±649.76)、(20 787.90±2 200.24)、(406.99±167.99)增加至(7 685.06±1 170.69)、(30 499.43±9 013.76)、(2 692.38±516.67)(n=3)。AC133的上调是转录水平和翻译后水平调控共同作用的结果。结论 AC133相对于CD133mRNA和蛋白更敏感地标记了胶质瘤干细胞。Objective To study the expression level of CD133 sensitively mirroring the phenotype of glioma stem cells.Methods The culture conditions of CD133 positive glioma stem cells which were established under serum-free stem cell permissive medium were changed from normoxia of 20% O2 into hypoxia of 1.5% O2 for 3 days.Real-time quantitative PCR was performed to study the expression of CD133 mRNA.Western blot was used to study the CD133 protein expression.Flow cytometry was performed to examine the extracellular and intracellular AC133 expression.ELISA was performed to assay the AC133 expression in the supernatant of medium.Results The immunofluorescence intensities of AC133 protein expression on the surface of CD133 positive glioma stem cells NCH421k,NCH441 and NCH440 under hypoxia were（7 685.06±1 170.69）,（30 499.43±9 013.76）,（2 692.38±516.67）（n=3）,which were significantly up-regulated when compared to under normoxia[（2 381.07±649.76）,（20 787.90±2 200.24）,（406.99±167.99）,n=3,P〈0.05].The up-regulation of AC133 was due to the change both on transcriptional and post-translational levels.Conclusion AC133 more sensitively mirrors glioma stem cell phenotype than CD133 mRNA and protein.

关 键 词：CD133 AC133 脑胶质瘤 脑肿瘤干细胞 缺氧 

分 类 号：R739.4[医药卫生—肿瘤]