脑源性神经营养因子促进人脑胶质瘤细胞株生长的研究

Effects of brain derived neurotrophic factor on growth of human glioma cell line

目的:探讨脑源性神经营养因子(BDNF)在人脑胶质瘤U87-MG细胞株生长中的作用及其可能的促血管生成机制。方法:脑胶质瘤U87-MG细胞株给予BD-NF蛋白刺激或BDNF抗体处理,采用realtime PCR检测血管内皮生长因子(VEGF)mRNA的表达,采用Transwell小室侵袭试验检测细胞侵袭能力,采用Annexin-Ⅴ/PI双染流式细胞术分析细胞凋亡。结果:BD-NF蛋白刺激后VEGFmRNA表达随着时间的延长而上调,与对照组相比,8h时增高了2.1倍,16h时增高了2.5倍。细胞侵袭力增加,紫色细胞计数对照组为317±16.1,8h为419±12.5,16h为511±10.1,差异有统计学意义,P<0.01。细胞凋亡率8h为2.28%,16h为2.04%,差异无统计学意义,P>0.05。BDNF抗体作用组与之相反,VEGFmRNA表达降低,8h时降低了0.88倍,16h时降低了0.78倍。紫色细胞计数8h为212±18,16h为195±17.7。细胞凋亡率8h为22.4%,16h为29%,差异有统计学意义,P<0.01。结论:BDNF在体外能够上调VEGFmR-NA表达,促进细胞侵袭,抑制细胞凋亡。

OBJECTIVE：To approach the effect of brain derived neurotrophic factor on human glioma cell line growth and probable pro-angiogenesis.METHODS：A human glioma U87-MG cell line was treated with BDNF protein or BDNF antibody.Real time PCR was used to detect the expression of VEGF mRNA,transwell chamber invasion experiment was used to check cell invasion ability,and flow cytometry Annexin-Ⅴ/P I double stain was used to analyze cell apoptosis.RESULTS：BDNF protein stimulation up-regulated the expression of VEGF mRNA as the time prolonged,the expression increased 2.1 fold in 8 h and 2.5 fold in 16 h,respectively.Purple cell counts were 419±12.5 in 8 h and 511±10.1 in 16 h,respectively,and were significantly different from the control group（317±16.1）as the cell invasion ability increased（P0.01）.There was no significant difference between the experimental groups and the control group（P0.05）.The apoptosis rates were 2.28% in 8 h and 2.04% in 16 h,with significant difference（P0.05）.While BDNF antibody had the opposite effect,the expression of VEGF mRNA decreased 0.88 fold in 8h and 0.78 fold in 16 h,respectively.Purple cell counts were 212±18 in 8 h and 195±17.7 in 16 h,respectively.The cell apoptosis rates were 22.4% in 8 h and 29% in 16 h respectively and was significantly different compared with the control group（P0.01）.CONCLUSION：BDNF can up-regulate the expression of VEGF mRNA,increase cell invasion and suppress cell apoptosis in vitro.