摘要
目的探讨RhoE对转录因子NF-κB活性水平的影响。方法将包含RhoE基因的质粒转染胶质瘤U87细胞;免疫印迹实验分析NF-κB亚单位p65在胞浆和胞核中的蛋白表达水平;细胞免疫化学实验检测p65在细胞中的蛋白表达水平和亚细胞定位;免疫印迹实验检测Ikkβ和IκBα蛋白表达水平。结果将pC MVFlA G-RhoE质粒成功转染U87细胞后,免疫印迹实验显示,胞浆和胞核中p65的蛋白表达水平均较对照组下降;细胞免疫化学结果亦表明,p65在细胞核中的定位量明显减少。免疫印迹实验进一步分析显示,NF-κB上游调控分子IκBα表达增加,而Ikkβ表达减少。结论上调RhoE表达水平能够抑制U87细胞转录因子NF-κB活性水平。NF-κB活性的降低可能与RhoE调控Ikk/IκBα机制相关。
Objective To investigate the effects of RhoE in regulating NF-κB activity.Methods Full-length RhoE vector was transiently transfected into U87 cells,protein level of p65,a sub structural unit of NF-κB,in both cytosol and nucleus was determined by western blot.Immuocytochemistry was performed to determine protein level and sub-cellular location of p65.Western blot was also used to detect Ikkβ and IκBα protein levels.Results As western blot showed,protein level of RhoE in both cytosol and nucleus was decreased significantly in U87 cells infected by pCMV-F1AG-RhoE vector successfully,compared with control,results from immuocytochemistry confirmed decrease of p65 protein level in nucleus.Western blot further indicated expression of one upstream regulator IκBα was increased,but Ikkβ decreased.Conclusions RhoE up-regulation could inhibit activity of NF-κB in glioma U87 cells.Inhibition of NF-κB activity is related to RhoE-regulated Ikk/IκBα.
出处
《临床神经外科杂志》
CAS
2014年第1期46-48,共3页
Journal of Clinical Neurosurgery
