[1]朱明亮,袁晖.RhoE表达上调对U87细胞NF-κB活性的影响[J].临床神经外科杂志,2014,(01):46-48.
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RhoE表达上调对U87细胞NF-κB活性的影响
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《临床神经外科杂志》[ISSN:1672-7770/CN:32-1727/R]

卷:
期数:
2014年01期
页码:
46-48
栏目:
论著
出版日期:
2014-02-28

文章信息/Info

Title:
RhoE up-regulation inhibits NF-κB activity in glioma U87 cells
作者:
朱明亮袁晖
解放军第169医院
关键词:
RhoE胶质瘤 NF-κB
Keywords:
RhoE Glioma NF-κB
分类号:
R739
文献标志码:
A
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 pCMV-FlAG-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(P<0.01);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(P<0.05)。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(P<0.01),而Ikkβ表达减少(P<0.01)。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。
Abstract:
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-FlAG-RhoE vector successfully, compared with control (P<0.01); results from immuocytochemistry confirmed decrease of p65 protein level in nucleus (P<0.05); western blot further indicated expression of one upstream regulator IκBα was increased (P<0.01), but Ikkβ decreased (P<0.01). 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.

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备注/Memo

备注/Memo:
[收稿日期] 2013-01-15
更新日期/Last Update: 2014-02-15