Glioblastoma stem cell differentiation into endothelial cells evidenced through live-cell imaging
Glioblastoma stem cell differentiation into endothelial cells evidenced through live-cell imaging
复制标题
通过活细胞成像证明胶质母细胞瘤干细胞分化为内皮细胞
DOI：
10.1093/neuonc/nox016
复制
发表时间：
2017-08-01
期刊：
影响因子：
15.9
通讯作者：
Chen, Zhong-Ping
中科院分区：
文献类型：
--
作者：
Mei, Xin;Chen, Yin-Sheng;Chen, Zhong-Ping
背景。胶质母细胞瘤细胞引发的血管形成是一种称为血管生成模拟的替代血管生成。然而，目前关于胶质母细胞瘤干细胞（GSCs）新生血管形成机制的知识有限。采用免疫荧光染色法检测64例患者胶质母细胞瘤标本和10例荧光胶质瘤异种移植标本内皮标志物（CD34和CD31）和胶质细胞标志物（胶质原纤维酸性蛋白[GFAP]）的表达。然后从人胶质母细胞瘤组织中分离gsc，建立CD133+/Sox2+含红色荧光蛋白(RFP)-GSC-1细胞。这些细胞形成血管结构的能力通过三维培养的活细胞成像来检测。64例临床胶质母细胞瘤样本中有30例（46.9%）存在CD34-GFAP或CD31-GFAP共表达胶质母细胞瘤源性内皮细胞（GDEC）。在这30个样本中，有21个（70%）样本发现GDEC形成血管结构。在21例GDEC血管样本中，CD34+ GDEC血管和CD31+ GDEC血管分别占血管总数的14.16%和18.08%。在异种移植样本中，10只小鼠中有7只小鼠检测到CD34+ GDEC， 7只小鼠中有4只存在CD34+ GDEC血管。7只小鼠有CD31+ GDEC， 4只小鼠有CD31+ GDEC血管（共10只）。通过活细胞成像，我们观察到部分胶质瘤细胞与血管内皮生长因子一起培养后CD34的表达逐渐增加，并记录了体外RFP-GSC-1内皮标记物表达的动态增加。培养6小时后，细胞表达CD34（9.46%）。结果表明，GSCs可在胶质母细胞瘤中分化为内皮细胞，促进血管生成。
Background. Glioblastoma cell-initiated vascularization is an alternative angiogenesis called vasculogenic mimicry. However, current knowledge on the mechanism of de novo vessel formation from glioblastoma stem cells (GSCs) is limited.Methods. Sixty-four glioblastoma samples from patients and 10 fluorescent glioma xenograft samples were examined by immunofluorescence staining for endothelial marker (CD34 and CD31) and glial cell marker (glial fibrillary acidic protein [GFAP]) expression. GSCs were then isolated from human glioblastoma tissue and CD133+/Sox2+ red fluorescent protein-containing (RFP)-GSC-1 cells were established. The ability of these cells to form vascular structures was examined by live-cell imaging of 3D cultures.Results. CD34-GFAP or CD31-GFAP coexpressing glioblastoma-derived endothelial cells (GDEC) were found in 30 of 64 (46.9%) of clinical glioblastoma samples. In those 30 samples, GDEC were found to form vessel structures in 21 (70%) samples. Among 21 samples with GDEC vessels, the CD34+ GDEC vessels and CD31+ GDEC vessels accounted for about 14.16% and 18.08% of total vessels, respectively. In the xenograft samples, CD34+ GDEC were found in 7 out of 10 mice, and 4 out of 7 mice had CD34+ GDEC vessels. CD31+ GDEC were also found in 7 mice, and 4 mice had CD31+ GDEC vessels (10 mice in total). Through live-cell imaging, we observed gradual CD34 expression when cultured with vascular endothelial growth factor in some glioma cells, and a dynamic increase in endothelial marker expression in RFP-GSC-1 in vitro was recorded. Cells expressed CD34 (9.46%) after 6 hours in culture.Conclusions. The results demonstrated that GSCs may differentiate into endothelial cells and promote angiogenesis in glioblastomas.