文章摘要
黄 为,徐 标,Wang Xuejun.GFPu-1克隆细胞反映泛素蛋白酶体功能[J].南京医科大学学报,2007,(2):126~128
GFPu-1克隆细胞反映泛素蛋白酶体功能
Establishment of GFPu-1 clonal cells as an indicator of ubiquitin-proteasome system
投稿时间:2006-08-11  
DOI:10.7655
中文关键词: 泛素蛋白酶体系统  绿色荧光蛋白  细胞
英文关键词: ubiquitin-proteasome system  green fluorescent protein  cells
基金项目:
作者单位
黄 为 南京大学医学院鼓楼医院心脏科江苏 南京 210008 
徐 标 南京大学医学院鼓楼医院心脏科江苏 南京 210008 
Wang Xuejun Caridovascular Research Institute, University of South Dakota, Sioux Falls, SD57105, USA 
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中文摘要:
      目的:建立反映泛素蛋白酶体系统(ubiquitin-proteasome system,UPS)功能的GFPu-1细胞系。方法: 将GFPu质粒,转染HEK293细胞,通过G418筛选培养出含GFPu的克隆细胞系GPPu-1,加入蛋白酶抑制剂 lactacystin,通过共聚焦显微镜和Western 印迹观察绿色荧光蛋白(GFP)的表达和分布。结果:加入lactacystin 后,GFPu转染细胞荧光强度明显增强,GFPu在胞浆和细胞核分布均匀,且GFPu和β-catenin明显积聚,GFPu增加了约2.5倍(P < 0.05),β-catenin增加了约1.2倍(P < 0.05)。结论:GFPu-1克隆细胞是一种反映UPS功能的敏感指标。
英文摘要:
       Objective:To establish GFPu-1 clonal cells reflecting the function of the ubiquitin-proteasome system. Methods:GFPu plasmid was transfected into human embryonic kidney(HEK) 293 cells. GFPu-1 cells with stable expression of GFPu were selected by G418. After proteasome inhibitor lactacystin was added into GFPu-1 cells, the distribution and expression of green fluorescent protein(GFP) were studied by confocal microscopy and Western blot. Results:GFPu was distributed diffusely in the nuclear and cytoplasmic compartment of GFPu-1 cells with lactacystin. Compared with the control cells with DSMO, the fluorescence of GFPu significantly increased in GFPu-1 cells with lactacystin. Accummuation of GFPu rose about 2.5 folds(P < 0.05) and β-catenin expression rose about 1.2 fold(P < 0.05). Conclusion:GFPu-1 clonal cells may be a sensitive indicator for the function of the ubiquitin-proteasome system.
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