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Transcriptional roles of nuclear factor κB and nuclear factor-interleukin-6 in the tumor necrosis factor α-dependent induction of cyclooxygenase-2 in MC3T3-E1 cells.
Open accessAbstract
When a mouse osteoblastic cell line MC3T3-E1 was cultured in the presence of tumor necrosis factor alpha (TNF alpha), the release of prostaglandin E2 and the cyclooxygenase activity increased in a dose- and time-dependent manner. The increase of the enzyme activity was attributed mostly to the induction of cyclooxygenase-2 rather than cyclooxygenase-1 as judged by the inhibitory effect of NS398, Western blotting, and Northern blotting. In this system we attempted to elucidate the transcriptional regulation of the cyclooxygenase-2 gene. As examined by the luciferase assay, two positive regulatory regions (-186 to -131 and -512 to -385 base pairs) were found in the 5'-flanking promoter region of the mouse cyclooxygenase-2 gene in the TNF alpha-stimulated cells. The former included putative NF-IL6 (C/EBP beta) and AP2 elements, and the latter contained the NF kappa B motif. A DNA probe including the NF-IL6 and AP2 sites gave positive bands upon electrophoretic mobility shift assay using the nuclear extracts of MC3T3-E1 cells. The bands were supershifted by the addition of anti-NF-IL6 antibody but not by anti-AP2 antibody. A probe including the NF kappa B site also gave positive bands, which were supershifted by anti-NF kappa B p50 and p65 antibodies. Furthermore, when the motif of NF-IL6 or NF kappa B or both was subjected to point mutation, the luciferase activity was markedly reduced. These data suggested a potential role of both NF-IL6 and NF kappa B in the induction of cyclooxygenase-2 by TNF alpha.
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“One of the conundrums is that IL-6 is remarkably noninflammatory in tissues, and although there are some reports that IL-6 induces PGs in non-CNS cells, this seems to be uncommon and to occur at quite high concentrations (57) despite the presence of a nuclear factor IL-6 site upstream of the COX-2 gene (58).”
From Interleukin-6 is an afferent signal to the hypothalamo-pituitary-adrenal axis during local inflammation in mice (2003) · cites this paper as [58]
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Metadata
- Authors
- Yamamoto, K., Arakawa, T., Ueda, N., & Yamamoto, S.
- Affiliations
- Tokushima University (JP)
- Corresponding author
- Yamamoto, Kei
- Yamamoto, Shozo
- Venue
- Journal of Biological Chemistry
- Publisher
- Elsevier BV
- Published
- 1995-12
- Volume
- 270
- Issue
- 52
- Pages
- 31315-31320
- ISSN
- 0021-9258 (ISSN-L)
- OpenAlex
- W2002708977
- Cited by
- 647
- References
- 38
- Citations by year
- 2026: 2 · 2025: 2 · 2024: 7 · 2023: 2 · 2022: 3 · 2021: 17 · 2020: 8 · 2019: 11 · 2018: 12 · 2017: 8 · 2016: 10 · 2015: 8 · 2014: 10 · 2013: 15 · 2012: 14
- FWCI
- 17.65
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Journal of Biological Chemistry (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Funders
- Japan Foundation for Applied Enzymology
- Japan Research Foundation for Clinical Pharmacology
- Kissei
- Ono Pharmaceutical
- Japan Society for the Promotion of Science
- Daiichi Sankyo Company
- Topics
- Health Sciences › Medicine › Pharmacology (medical) › Inflammatory mediators and NSAID effects
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › NF-κB Signaling Pathways
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Signaling Pathways in Disease
- Keywords
- cyclooxygenase-2, NF-κB, NF-IL6, C/EBPβ, TNF-α, transcriptional regulation, prostaglandin E2, MC3T3-E1 cells, osteoblasts, promoter regulation, gene induction
- MeSH terms
- Animals, Base Sequence, DNA-Binding Proteins, Enzyme Induction, Isoenzymes, Molecular Sequence Data, Nuclear Proteins, Oligodeoxyribonucleotides, Promoter Regions, Genetic, Prostaglandin-Endoperoxide Synthases, Regulatory Sequences, Nucleic Acid, Transcription, Genetic, Tumor Necrosis Factor-alpha, NF-kappa B, 3T3 Cells, CCAAT-Enhancer-Binding Proteins, Mice
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