人脑胶质瘤干细胞分化为血管内皮细胞的实验研究  

Endothelial cells differentiation of human glioma stem/progenitor cells

作　　者：聂秀涛[1,2] 金贵善[1,2] 米蕊芳[1,2] 张国滨[1,2] 谢坚[1,2] 曹泽[1,2] 周益强[1,2] 董程远[1,2] 刘福生[1,2] 

机构地区：[1]首都医科大学北京市神经外科研究所,100050 [2]首都医科大学附属北京天坛医院100050

出　　处：《中华神经外科杂志》2013年第3期289-293,共5页Chinese Journal of Neurosurgery

基　　金：国家自然科学基金（81071776）

摘　　要：目的探讨体内外原代培养的人脑胶质瘤干细胞（GSCs）与胶质瘤新生血管内皮细胞的关系。方法新鲜高级别（WHOⅢ级、Ⅳ级）的人脑胶质瘤标本经原代培养获取GSCs，免疫组化法检测其肿瘤干细胞及干细胞标记物Nestin的表达；鉴定后的GSCs经低氧诱导，免疫荧光法检测其诱导分化后内皮细胞标记物CD31、CD144和胶质瘤细胞标记物GFAP的表达，RT—PCR和Western—blot法检测其CD31的表达；建立胶质瘤干细胞皮下荷瘤裸鼠模型，免疫组化技术检测模型中人来源的CD31的表达。结果（1）悬浮生长的胶质瘤干细胞球样细胞经免疫组化鉴定Nestin表达阳性；（2）低氧诱导后的GSCs能够表达CD31、CD144，有些细胞能够同时表达CD144和GFAP；（3）RT—PCR检测发现GSCs在诱导前后都有CD31mRNA的表达，而Western—blot检测到只有诱导后的GSCs有CD31蛋白的表达；（4）胶质瘤干细胞荷瘤裸鼠模型的肿瘤组织中部分微血管抗人CD31抗体染色阳性。结论胶质瘤干细胞不仅在体外低氧条件下可分化为内皮细胞，在体内微环境条件下同样可分化为血管内皮细胞，并参与胶质瘤新生组织的血液供应。Objective With the development of glioblastoma therapy, the view on inhibiting angiogenesis had reached a general consensus, but the efficacy is not satisfactory. It is important to study the relationships between glioma stem cells （GSCs） and endothelial cells, which might provide a new therapeutic target to improve the prognosis of glioblastoma. Methods We isolated the GSCs from 38 specimens and cultured them as spheres in serum - free medium. After incubating in vitro for 4 h under hypoxia, we observed the expression of the endothelial cell markers CD31 and CD144 and glial fibrillary acidic protein （GFAP） by immunofluorescence staining, RT - PCR and Western - blotting. In the xenografts of transplanted tumors established by GSCs spheres, we detected vessels with an anti - human endothelial cell antiboy. Results CD31 and CD144 were expressed in transdifferentiated cells in vitro, as determined with immunofluorescence staining and Western - blot analysis. Additionally, at the mRNA level we found that both the GSCs and the transdifferentiated cells expressed the endothelial cell markers. Some micro-vessels in vivo were CD31 positive in the immunofluorescence staining. Conclusions demonstrates that GSCs have the ability to differentiate into endothelial cells. Our study

关 键 词：胶质瘤干细胞 内皮细胞 低氧诱导 

分 类 号：R739.4[医药卫生—肿瘤]