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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。
Objective: To observe the effect of ING4 on proliferation and migration of human glioma cell line U251 and to explore its possible mechanism. Methods: U251 cells were transfected with Ad-ING4 and adenovirus empty vector. The expression of ING4 gene was detected by RT-PCR and the expression of the target protein was detected by Western blot. The effect of ING4 on the proliferation of U251 cells was detected by MTT assay. The influence of ING4 on the invasiveness of U251 cells was detected by BoydenChamber test. The expressions of NGF and TrkA were detected by Western blotting. The expression of RhoA was detected by pull-down assay. RESULTS: After U251 cells were infected with Ad-ING4 for 48 h, RT-PCR results showed that ING4 was overexpressed in U251 cells and the proliferation and migration of U251 cells were significantly inhibited. Immunoblotting showed that the expression of TrkA and NGF in U251 cells infected with AdING4 was decreased. The results of pull-down assay showed that the expression of active RhoA was decreased. Conclusion: Ad-ING4 can inhibit the proliferation and migration of glioma cell line U251, and this effect may be through inhibiting the expression of NGF, TrkA and active RhoA.