NDLI: Establishment of a cell-based assay for examining the expression of tumor necrosis factor alpha (TNF-α) gene

Content Provider	Springer Nature Link
Author	Chen, Qiang Zhao, Yang Cheng, Zhuo Xu, Yixiang Yu, Chundong
Copyright Year	2008
Abstract	Tumor necrosis factor alpha (TNF-α) is a proinflammatory cytokine produced by activated macrophages and lymphocytes and involved in many inflammatory diseases. Preventing the production or action of TNF-α is a potent therapeutic strategy for these inflammatory diseases. Since there is a lack of rapid and effective assay for examining the expression TNF-α in macrophages, we attempt to establish a reporter system to assess TNF-α gene expression through measuring luciferase activity. In this study, mouse macrophage cell line RAW 264.7 was stably transfected with a luciferase reporter pGL3-TNFPro-UTR, which contains TNF-α promoter and 3′-untranslated region (3′-UTR). The TNF-α-luciferase reporter cell line is used for assessing the expression of TNF-α gene induced by LPS in the presence or absence of chemicals that inhibit the biosynthesis of TNF-α such as dexamethasone and emodin, and also for measuring change of expression of TNF-α gene under downregulation of the expression of steroid receptor coactivator-3, a modulator for TNF-α. The luciferase activity correlated well with the ELISA results for TNF-α production, therefore, the TNF-α-luciferase reporter cell line is a sensitive, effective tool for studying the expression of TNF-α gene.
Starting Page	357
Ending Page	363
Page Count	7
File Format	PDF
ISSN	01757598
Journal	Applied Microbiology and Biotechnology
Volume Number	80
Issue Number	2
e-ISSN	14320614
Language	English
Publisher	Springer-Verlag
Publisher Date	2008-06-17
Publisher Place	Berlin, Heidelberg
Access Restriction	Subscribed
Subject Keyword	TNF-α Luciferase activity Macrophage LPS Microbial Genetics and Genomics Microbiology Biotechnology
Content Type	Text
Resource Type	Article
Subject	Medicine Applied Microbiology and Biotechnology Biotechnology

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