Interleukin-1 beta and dexamethasone regulate gene expression of prostaglandin H synthase-2 via the NF-kB pathway in human amnion derived WISH cells.
Interleukin-1beta (IL-1beta) stimulated PGE2 synthesis in human amnion derived WISH cells, whereas dexamethasone blocked IL-1beta-mediated stimulation of PGE2 production. Sequence analysis of the 5'-flanking region of the human prostaglandin H synthase-2 (PGHS-2) gene indicates two putative NF-kB binding sites. Mutation of a single site or both sites resulted in significantly decreased activity of the PGHS-2 promoter. IL-1beta treatment increased significantly the native promoter activity and this increase was attenuated by using the NF-kB-mutant promoter. Dexamethasone treatment also decreased the IL-1beta mediated stimulation of the PGHS-2 native promoter but not the NF-kB mutant promoter. Furthermore, the involvement of the NF-kB was supported by electrophoretic mobility shift assay which revealed an increased nuclear binding of the NF-kB probe upon IL-1beta induction and a decreased nuclear binding of the NF-kB probe upon dexamethasone pre-treatment. These results provide convincing evidence that NF-kB may mediate the IL-1beta stimulation of PGHS-2 gene expression as well as the dexamethasone inhibition of the IL-1beta induction process in WISH cells.
Division of Medicinal Chemistry and Pharmaceutics, College of Pharmacy, The University of Kentucky, Lexington 00536-0082, USA.
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D020121:5' Untranslated Regions / Q000737:chemistry / Q000187:drug effects / Q000235:genetics; D000650:Amnion / Q000166:cytology / Q000378:metabolism*; D001665:Binding Sites / Q000235:genetics; D002478:Cells, Cultured; D051546:Cyclooxygenase 2; D003907:Dexamethasone / Q000494:pharmacology*; D005786:Gene Expression Regulation / Q000187:drug effects*; D006801:Humans; D007375:Interleukin-1 / Q000494:pharmacology*; D007527:Isoenzymes / Q000235:genetics*; D008565:Membrane Proteins; D016328:NF-kappa B / Q000378:metabolism / Q000502:physiology; D010544:Peroxidases / Q000235:genetics*; D011451:Prostaglandin-Endoperoxide Synthases / Q000235:genetics*; D014157:Transcription Factors / Q000378:metabolism
D020121:5' Untranslated Regions; D007375:Interleukin-1; D007527:Isoenzymes; D008565:Membrane Proteins; D016328:NF-kappa B; D014157:Transcription Factors; D003907:Dexamethasone; D010544:Peroxidases; D051546:Cyclooxygenase 2; C496540:PTGS2 protein, human; D011451:Prostaglandin-Endoperoxide Synthases