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OtherIMMUNOPHARMACOLOGY

Induction of Prostaglandin H Synthase-2 and Tumor Necrosis Factor-α in Human Amnionic WISH Cells by Various Stimuli Occurs Through Distinct Intracellular Mechanisms

Keren I. Hulkower, Ellen R. Otis, Junling Li, Bruce W. Ennis, David J. Cugier, Randy L. Bell, George W. Carter and Keith B. Glaser
Journal of Pharmacology and Experimental Therapeutics February 1997, 280 (2) 1065-1074;
Keren I. Hulkower
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Ellen R. Otis
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Junling Li
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Bruce W. Ennis
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David J. Cugier
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Randy L. Bell
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George W. Carter
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Keith B. Glaser
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Abstract

These studies examined the signal transduction mechanisms by which prostaglandin (PG) E2 production can occur in human amnionic WISH cells in response to the stimuli okadaic acid, interleukin (IL)-1β, tumor necrosis factor (TNF)-α, phorbol-12-myristate-13-acetate (PMA) or combinations of PMA with IL-1β or TNF-α. We also investigated whether WISH cells are capable of producing TNF-α or IL-1β in response to stimulation, because these cytokines can be produced in an autocrine fashion to perpetuate an inflammatory response. Our data indicate that the magnitude of PGE2 production induced by a given stimulus correlated temporally with the level of PGH synthase-2 (PGHS-2) protein. PMA or IL-1β induced PGE2 production 2 to 4 hr after treatment, whereas the combination of these agents produced the most rapid induction 2 hr after treatment. Only okadaic acid induced the production of both PGE2 and TNF-α, after a lag of 12 to 18 hr. PGE2 production by all stimuli was inhibited by dexamethasone, the IL-1 receptor antagonist (IL-1ra), the specific PGHS-2 inhibitor NS-398 and the protein kinase inhibitor staurosporin. In contrast, TNF-α production in response to okadaic acid was inhibited by the TNF-converting enzyme inhibitor GI 129471 and staurosporin but was unaffected by either IL-1ra, dexamethasone or NS-398. We conclude that WISH cells are capable of producing bioactive proinflammatory mediators such as TNF-α and PGE2 through separable intracellular signal transduction mechanisms. The ability of IL-1ra to reduce PGE2 production caused by all stimuli used suggests an autocrine role for IL-1 in PGHS-2 induction in these cells.

Footnotes

  • Send reprint requests to: Dr. Keren I. Hulkower, Abbott Laboratories, D47K/AP9, 100 Abbott Park Road, Abbott Park, IL 60064-3500.

  • Abbreviations:
    EIA
    enzyme immunoassay
    ELISA
    enzyme-linked immunosorbent assay
    IL
    interleukin
    IL-1ra
    interleukin-1 receptor antagonist
    PG
    prostaglandin
    PGHS
    prostaglandin H synthase
    PLA2
    phospholipase A2
    PMA
    phorbol-12-myristate-13-acetate
    SDS
    sodium dodecyl sulfate
    SSC
    standard saline citrate
    TBS
    Tris-buffered saline
    TNF
    tumor necrosis factor
    • Received June 7, 1996.
    • Accepted October 7, 1996.
  • The American Society for Pharmacology and Experimental Therapeutics
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Journal of Pharmacology and Experimental Therapeutics
Vol. 280, Issue 2
1 Feb 1997
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OtherIMMUNOPHARMACOLOGY

Induction of Prostaglandin H Synthase-2 and Tumor Necrosis Factor-α in Human Amnionic WISH Cells by Various Stimuli Occurs Through Distinct Intracellular Mechanisms

Keren I. Hulkower, Ellen R. Otis, Junling Li, Bruce W. Ennis, David J. Cugier, Randy L. Bell, George W. Carter and Keith B. Glaser
Journal of Pharmacology and Experimental Therapeutics February 1, 1997, 280 (2) 1065-1074;

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OtherIMMUNOPHARMACOLOGY

Induction of Prostaglandin H Synthase-2 and Tumor Necrosis Factor-α in Human Amnionic WISH Cells by Various Stimuli Occurs Through Distinct Intracellular Mechanisms

Keren I. Hulkower, Ellen R. Otis, Junling Li, Bruce W. Ennis, David J. Cugier, Randy L. Bell, George W. Carter and Keith B. Glaser
Journal of Pharmacology and Experimental Therapeutics February 1, 1997, 280 (2) 1065-1074;
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