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目的观察下调胶质瘤干细胞CXCR4表达后对其侵袭能力和血管内皮生长因子(vascular endothelial growth factor,VEGF)分泌量的影响。方法采用间接免疫荧光标记观测U87细胞及其颅内移植瘤标本中胶质瘤干细胞标记物CD133和CXCR4共表达情况;分别从稳定转染CXCR4小干扰RNA质粒的U87细胞和稳定转染阴性对照质粒的U87细胞中培养出胶质瘤干细胞球,并对其进行鉴定,观察两者CXCR4的表达情况;通过Transwell小室比较两者的侵袭能力;采用酶联免疫吸附实验(ELISA)检测两者细胞培养上清内VEGF的含量。结果U87细胞及其移植瘤中均可检测到CXCR4与CD133共表达,下调胶质瘤干细胞CXCR4表达后,胶质瘤干细胞的体外侵袭能力明显下降[干扰组:(17.0±5.8),对照组:(71.5±8.7),P<0.05],VEGF分泌明显减少[干扰组:24 h(690±24)pg/ml,48 h(1 504±53)pg/ml;对照组:24 h(850±25)pg/ml,48 h(2 001±150)pg/ml,P<0.05]。结论胶质瘤干细胞表达CXCR4,下调其表达可抑制胶质瘤干细胞的侵袭和促血管生成能力,提示CXCR4可能成为针对胶质瘤干细胞进行治疗的靶点。
Objective To investigate the effect of CXCR4 on the invasion and the secretion of vascular endothelial growth factor (VEGF) in glioma stem cells. Methods Indirect immunofluorescence staining was used to observe the co-expression of glioma stem cell markers CD133 and CXCR4 in U87 cells and their intracranial xenografts. U87 cells stably transfected with CXCR4 small interfering RNA plasmid and stable transfected negative control plasmid Of U87 cells cultured glioma stem cell spheres, and its identification, to observe the expression of both CXCR4; Transwell chambers were compared by the invasion ability; enzyme-linked immunosorbent assay (ELISA) both cell culture The content of VEGF in the supernatant. Results Both CXCR4 and CD133 were co-expressed in U87 cells and their xenografts. The down-regulation of CXCR4 expression in glioma stem cells decreased the invasiveness of glioma stem cells in vitro [Interference group: (17.0 ± 5.8) vs control group (71.5 ± 8.7), P <0.05], and the secretion of VEGF was significantly decreased [6 hours (690 ± 24) pg / ml and 48 hours 25) pg / ml for 48 h (21 001 ± 150) pg / ml, P <0.05]. Conclusion The expression of CXCR4 in glioma stem cells is down-regulated. The downregulation of glioma stem cells may inhibit the invasion and angiogenesis of glioma stem cells, suggesting that CXCR4 may be a target of treatment for glioma stem cells.