摘要
背景与目的:在恶性胶质瘤中存在一类恶性度极高的前体细胞,它们控制着肿瘤的生长与恶性演进,也是肿瘤复发的根源。本研究对胶质母细胞瘤进行肿瘤干细胞的分离与恶性行为特征分析,旨在验证胶质瘤干细胞的存在,并为未来该领域研究打下基础。方法:充分消化术中切除的胶质母细胞瘤组织块至单细胞悬液,流式细胞术检测CD133+肿瘤细胞百分率,免疫磁珠分选法分离CD133+和CD133-胶质母细胞瘤细胞;免疫荧光法检测两类细胞中巢蛋白(nestin)、胶质纤维酸性蛋白(GFAP)和神经元特异性烯醇化酶(NSE)的表达水平;Transwell双室培养体系检测两类瘤细胞的侵袭能力,血清依赖实验检测两类瘤细胞在低营养条件下的生存能力,软琼脂克隆形成实验检测瘤细胞的克隆形成能力;去胸腺小鼠腹股沟皮下接种瘤细胞,观察成瘤率。结果:流式细胞术检测CD133+肿瘤细胞百分率为(2.31±0.57)%。CD133+细胞高表达nestin,低表达GFAP和NSE;CD133-细胞则相反。与CD133-细胞比较,CD133+细胞具有更高的细胞侵袭能力、低营养耐受能力和克隆形成能力。CD133+细胞在小鼠体内成瘤率为87%(26/30),CD133-细胞为7%(2/30)。结论:CD133+胶质母细胞瘤细胞具有更高的恶性细胞表型,提示其具有高度研究价值。
BACKGROUND ＆ OBJECTIVE： CD133-positive cells in glioma have been reported to be associated with the malignant phenotype. In this study we isolated CD133-positive glioblastoma cells and analyzed its malignant performance characteristics. METHODS： The single cell suspension was prepared from glioblastoma tissue, the percentage of CD133 positive cells was detected by flow cytometry, then the CD133 positive and negative cells were isolated by immunomagnetic beads. The express level of nestin, glial fibrillary acidic protein （GFAP） and neurone specitlc enolase （NSE） were detected by immunofluorescenee. The invasion ability of CD133 positive and negative cells was compared by transwell assay, then the survival ability was detected by blood-serum deprive culture.detected in soft agar. Finally, transplanted into the by clone formation these cells were subeutaneous of immonodeficient mice and the tumor form rate was surveyed. RESULTS： The percentage of CD133 positive cells was （2.31_＋0.57）%. The CD133 positive cells expressed higher level ofnestin protein and lower level of GFAP and NSE, but the CD133 negative cells were opposite. The CD133 positive cells had much stronger invasion ability, survival ability in low nutrition circumstances, and the clonality than the CD133 negative cells. The tumor form rate of CD133 positive cells was 87% （26/30）, but the CD133 negative cells was 7% （2/30）. CONCLUSION： The CD133 positive glioblastoma ceils have more malignant performance characteristics than the CD133 negative ceils.
出处
《中国神经肿瘤杂志》
2012年第2期84-89,共6页
Chinese Journal of Neuro-Oncology
基金
国家自然科学基金项目(81000901)
天津市卫生局科技项目(2011KZ24)
