摘要
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 observe the effects of CXCR4 downregulation on the invasion capacity and VEGF secretion of glioma stem cells （GSCs）. Methods Coexpression of CXCR4 and CD133 in U87 cells and their xenografts grown in nude mice brain was detected by indirect immunofluorescence, under a laser scanning microscope. Tumorspheres were isolated from control U87 cells and CXCR4-siRNA U87 cells, respectively. We investigated the expression of CD133, nestin and CXCR4 in tumorspheres by laser scanning of CXCR4 downregulation on invasion and VEGF production of GSCs were detected by ELISA respectively. Results CXCR4 and CD133 were coexpressed in U87 cells and thei microscopy. Effects transwell assay and r xenografts. Down- regulation of CXCR4 expression inhibits the invasion capacity [ siRNA ： （ 17.0 ± 5.8 ）, control ： （ 71.5 ±8.7 ）, P 〈 0. 05 ] and VEGF secretion of GSCs [ siRNA ： 24 h （690± 24） pg/ml,48 h （ 1 504 ± 53 ） pg/ml, control ： 24 h （850±25）pg/ml,48 h （2 001 ± 150） pg/ml, P 〈0. 051. Conclusion GSCs express CXCR4, and downregulation of CXCR4 expression attenuates the invasion and pro-angiogenic capacity of GSCs, which suggests that CXCR4 expressed on GSCs may be a potential target of glioma therapy.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2009年第11期1049-1052,共4页
Journal of Third Military Medical University
