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学科主题基础医学
Establishment of a T-Ag and NF-kappa B binding elements gene co-transfected stable HUVECs cell line
Liu, YX; Cao, WS; Irani, K; Li, XJ
关键词Cell Line Co-transfection T-ag Nf-kappa b Reporter Gene Activity Assay
刊名JOURNAL OF CELLULAR BIOCHEMISTRY
2003-08-15
DOI10.1002/jcb.10588
89期:6页:1285-1290
收录类别SCI
文章类型Article
WOS标题词Science & Technology
类目[WOS]Biochemistry & Molecular Biology ; Cell Biology
研究领域[WOS]Biochemistry & Molecular Biology ; Cell Biology
英文摘要

To set up a T-Ag gene transfected stable human umbilical venous endothelial cells (HUVECs) cell line and a T-Ag, nuclear transcriptional factor kappa B (NF-kappaB) binding elements linked with luciferase reporter gene co-transfected stable HUVECs cell line. Cultured HUVECs were transfected with pCl-neo-T-Ag and pRSV-luc-3XkappaB by lipofectin. The G418 selected monoclones were subcultured. The expression of marker protein, vWF and the characteristic of uptake of lipids were compared by Western blotting and immunocytochemistry in non-transfected and transfected HUVECs. The reporter gene assay was done in the presence of TNF-alpha. A T-Ag gene transfected stable HUVECs cell line and a T-Ag and NF-kappaB binding elements linked with luciferase reporter gene co-transfected stable HUVECs cell lines were set up. The expression of vWF of these cell lines was similar with those in non-transformed HUVECs. The function of uptaking of lipids was preserved as well in transfected cell lines. Furthermore, TNF-alpha, a typical cytokine increasing the activity of NF-kappaB was used to treat the transfected cells O/N. The higher luciferase reporter gene activity was seen. A pCl-neo-T-Ag and pRSV-luc-3X kappaB co-transfected stable HUVECs cell line might be used to check reporter gene activity directly. It might be a useful tool to screen drugs acting on transcription level. (C) 2003 Wiley-Liss, Inc.

语种英语
WOS记录号WOS:000184609400018
引用统计
被引频次:1[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://ir.bjmu.edu.cn/handle/400002259/53672
专题北京大学基础医学院_药理学系
作者单位1.Peking Univ, Dept Pharmacol, Sch Basic Med Sci, Hlth Sci Ctr, Beijing 100083, Peoples R China
2.Johns Hopkins Univ Hosp, Sch Med, Div Cardiol, Baltimore, MD 21205 USA
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Liu, YX,Cao, WS,Irani, K,et al. Establishment of a T-Ag and NF-kappa B binding elements gene co-transfected stable HUVECs cell line[J]. JOURNAL OF CELLULAR BIOCHEMISTRY,2003,89(6):1285-1290.
APA Liu, YX,Cao, WS,Irani, K,&Li, XJ.(2003).Establishment of a T-Ag and NF-kappa B binding elements gene co-transfected stable HUVECs cell line.JOURNAL OF CELLULAR BIOCHEMISTRY,89(6),1285-1290.
MLA Liu, YX,et al."Establishment of a T-Ag and NF-kappa B binding elements gene co-transfected stable HUVECs cell line".JOURNAL OF CELLULAR BIOCHEMISTRY 89.6(2003):1285-1290.
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