摘要
目的:观察ING4对人脑胶质瘤细胞株U251增殖及迁移的影响,并探讨其可能的作用机制。方法:以Ad-ING4及腺病毒空载体转染U251细胞,RT-PCR法检测ING4基因的表达水平,Western blot法检测目的蛋白的表达。用MTT试验检测ING4对于U251细胞增殖的影响,通过BoydenChamber试验检测其对U251细胞侵袭能力的影响。并用免疫印迹方法检测NGF、TrkA的表达变化,通过pull-down试验检测活性RhoA表达。结果:U251细胞感染Ad-ING4 48 h后,RT-PCR结果显示ING4在U251中过表达,且U251细胞的增殖及迁移能力受到明显抑制。免疫印迹方法显示感染AdING4的U251细胞中TrkA和NGF的表达降低,pull-down试验结果显示活性RhoA表达降低。结论:Ad-ING4可以抑制胶质瘤细胞株U251的增殖和迁移,这一作用可能是通过抑制NGF、TrkA和活性RhoA的表达实现的。
AIM： To investigate the effect of Ad-ING4 on proliferation and migration of glioma cells and explore its probable mechanism. METHODS： U251 were infected with Ad-ING4. ING4 gene expression was evaluated by RT- PCR. MIT assay was adopted to evaluate the effect of ING4 on proliferation of U251; Boyden chamber assay was used to check the effect of ING4 on the migration of U251. In ING4 transfected U251, Western blot was used for detecting NGF and TrkA expression; Pull-down assay was used for detecting active RhoA expression. RESULTS： ING4 was overexpressed in Ad-ING4 transfected U251 cells. ING4 in- hibited proliferation and migration of U251 significantly. Moreover, overexpression of ING4 result in depression of NGF, TrkA and active RhoA. CONCLUSION： ING4 mediated inhibition of the proliferation and migration of human glioma cells by down regulating NGF, TrkA and active RhoA expression.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2011年第11期1188-1190,1194,共4页
Chinese Journal of Cellular and Molecular Immunology
基金
镇江市社会发展计划项目(SH2009018)
