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作 者:湛利平[1] 李巧玉[1] 张焱[1] 张铁军[1] 袁志诚[1] 陆培松[1]
机构地区:[1]江苏大学附属人民医院神经外科,江苏镇江212002
出 处:《细胞与分子免疫学杂志》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)
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.
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