胶质瘤干细胞血管生成拟态现象的体外研究

Vasculogenic Mimicry Assay of Glioma Stem Cell in Vitro

背景与目的:血管生成拟态是存在于恶性肿瘤中的,由肿瘤细胞而不是内皮细胞围成的管腔样结构,管腔中有血液流通并参与肿瘤微循环。我们采用体外三维培养模型来观察对比不同来源的胶质瘤干细胞血管生成拟态现象。方法:采用悬浮克隆球形成法诱导获得胶质瘤干细胞,应用胶质瘤干细胞相关分子标记免疫荧光技术鉴定及胶质瘤干细胞裸鼠移植成瘤试验证实已诱导获取的胶质瘤干细胞;采用体外三维培养模型来观察胶质瘤干细胞体外形成血管生成拟态现象。结果:成功获取6种不同来源的胶质瘤干细胞:GSC-1、GSC-2(来源于临床胶质母细胞瘤标本)和SKMG-4G、SKMG-1G、SF-295G、SF-767G(分别来源于胶质瘤细胞系SKMG-4、SKMG-1、SF-295、SF-767)。体外三维培养模型中,拟态管腔分圆形、多边形和三角形3种。GSC-1和SKMG-4G能形成典型的拟态管腔;SKMG-1G和SF-295G只能形成过渡性的非典型的拟态管腔;而GSC-2和SF-767G不能形成拟态管腔。结论:不同来源的胶质瘤干细胞拟态血管形成的能力不同,形态也各异。

BACKGROUND OBJECTIVE： Vasculogenic mimicry （VM） is known as non-endothelial tumor cell-lined microvascular channels in aggressive tumors, which may function as blood supply networks. In this study, three-dimensional （3D） matrigel tube formation assay was used to investigate the capacity of VM formation in different glioma stem-like cells （GSCs） in vitro. METHODS： GSCs were obtained from fresh surgical specimens and from glioma cell lines by means of serum-free clone formation, then immunofluorescence staining and a xenograft model was employed to detect the expression of differentiation markers for the multidifferentiation and oncogenicity. Three-dimensional matrigel culture was used to evaluate the ability of VM formation in different GSCs. RESULTS： Six GSC lines were successfully established：GSC-1 and GSC-2 derived from clinical glioblastoma specimens, and SKMG-4G,SKMG-1G,SF-295G and SF-767G derived from glioma cell lines SKMG-4,SKMG-1,SF-295 and SF-767,respectively. In 3-D matrigel culture, GSC-1 and SKMG-4G cells can growth and form close loops of round, polygon and triangle, which could be recognized as typical VM formation. While in SKMG-1G and SF-295G, only atypical VM formation （open circles） were observed. However, there was no any VM formation （neither close loop no open circle）found in GSC-2 and SF-767G. CONCLUSIONS： Glioma stem-like cells have different ability to form VM. And the morphology of VM in GSCs can be different.