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Development of a reporter assay system to screen chemicals for claudin‐4 modulator activity
Author(s) -
Kondoh Masuo,
Watari Akihiro,
Yagi Kiyohito
Publication year - 2012
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.26.1_supplement.848.10
Subject(s) - reporter gene , luciferase , chemistry , repressor , gene silencing , claudin , inducer , microbiology and biotechnology , gene , gene expression , biochemistry , transfection , biology , tight junction
Claudins (CLs), a tetra‐transmembrane protein family that contains 27 members, are key functional components of tight junction (TJ) seals. Modulation of CL‐4‐based TJ‐seals is a potent strategy for non‐invasive drug delivery. The barrier function of CL‐4 is disrupted in intestinal inflammation. These findings strongly suggest that CL‐4 modulators are promising candidates for CL‐4– targeted drug development; however, CL‐4 modulators have never been fully developed. In the present study, we developed a simple screening system for CL‐4 modulators by using a reporter gene. We cloned the promoter region of the CL‐4 gene, and prepared cells that stably expressed the luciferase reporter gene driven by the CL‐4 promoter. Treatment of the cells with a CL‐4 inducer or repressor resulted in an increase or decrease, respectively, in the luciferase activity in the cells. These findings suggest that the reporter‐expressing cells are a potent screening system for modulators of CL‐4 expression. We used the cells to screen 86 chemicals for CL‐4 modulator activity, and identified several CL‐4 modulator candidates. Potassium carbonate is a CL‐4 repressor; and curcumin, thiabendazole and carotene are CL‐4 inducers. We demonstrated that these four chemicals also modulate the TJ‐barrier. Thus, this simple reporter system will be a useful tool for the development of CL‐4 modulators.