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