Tumour necrosis factor-alpha regulation of prostaglandin H synthase-2 transcription is not through nuclear factor-kappa B in amnion-derived AV-3 cells

Hansen, W. R., Marvin, K. W., Potter, S. and Mitchell, M. D. (2000) Tumour necrosis factor-alpha regulation of prostaglandin H synthase-2 transcription is not through nuclear factor-kappa B in amnion-derived AV-3 cells. Placenta, 21 8: 789-798. doi:10.1053/plac.2000.0576


Author Hansen, W. R.
Marvin, K. W.
Potter, S.
Mitchell, M. D.
Title Tumour necrosis factor-alpha regulation of prostaglandin H synthase-2 transcription is not through nuclear factor-kappa B in amnion-derived AV-3 cells
Journal name Placenta   Check publisher's open access policy
ISSN 0143-4004
1532-3102
Publication date 2000-11
Sub-type Article (original research)
DOI 10.1053/plac.2000.0576
Volume 21
Issue 8
Start page 789
End page 798
Total pages 10
Place of publication London, England
Publisher W.B. Saunders
Language eng
Subject 1103 Clinical Sciences
1114 Paediatrics and Reproductive Medicine
Abstract Tumour necrosis factor (TNF)-α-stimulated prostaglandin (PG) E2biosynthesis by amnion-derived AV3 cells is accompanied by increased prostaglandin H synthase (PGHS)-2 mRNA expression. PGHS-1 mRNA expression is unchanged. PGHS-2 promoter–reporter constructs (−891/+9 and 5′ deletions thereof) were prepared. The regions containing concensus nuclear factor κB (NF-κB) elements (−447/−438 and −222/−213) did not enhance promoter activity. Elements associated with both basal and TNF-α-stimulated expression lie between bases −52 and −203. Site-directed mutagenesis of nuclear factor of interleukin-6 (NF-IL6) and cyclic AMP response elements (CREs) in this region reduced both basal and induced transcriptional activity of the −203/+9 construct by over 95 per cent. Electrophoretic mobility-shift assays using oligonucleotides derived from these sites demonstrated formation of specific DNA–protein complexes. Both NF-IL6 and CRE unlabelled oligonucleotides inhibited complex formation with the NF-IL6 oligonucleotide probe. Unlabelled CRE oligonucleotide also effectively inhibited formation of the complex with the CRE probe, but reduced effectiveness was observed when the NF-IL6 oligonucleotide was the competitor. Finally, unlabelled, consensus NF-κB oligonucleotide failed to compete for either probe. TNF-α treatment did not increase levels of these complexes. Thus NF-κB does not enhance basal or TNF-α-responsive PGHS-2 transcription in amnion-derived AV-3 cells. A permissive role for NF-IL6/CRE binding proteins in regulating PGHS-2 expression in these cells is indicated, but requires further clarification.
Keyword N-terminal Kinase
Preterm Labor
Messenger-rna
Human Trophoblasts
Gene-expression
Phorbol Ester
Wish Cells
Cyclooxygenase-2
Activation
infection
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status Unknown

Document type: Journal Article
Sub-type: Article (original research)
Collection: UQ Centre for Clinical Research Publications
 
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Created: Thu, 26 Aug 2010, 12:53:48 EST