title: Induction of prostaglandin H synthase-2 and tumor necrosis factor-alpha in human amnionic WISH cells by various stimuli occurs through distinct intracellular mechanisms.
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)-1beta, tumor necrosis factor (TNF)-alpha, phorbol-12-myristate-13-acetate (PMA) or combinations of PMA with IL-1beta or TNF-alpha. We also investigated whether WISH cells are capable of producing TNF-alpha or IL-1beta 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-1beta 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-alpha, 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-alpha 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-alpha 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.
authors.affiliation: Immunosciences Research Area, Abbott Laboratories, Abbott Park, Illinois 60064-3500, USA.
authors.forename: K I
authors.initials: KI
authors.lastname: Hulkower
authors.forename: E R
authors.initials: ER
authors.lastname: Otis
authors.forename: J
authors.initials: J
authors.lastname: Li
authors.forename: B W
authors.initials: BW
authors.lastname: Ennis
authors.forename: D J
authors.initials: DJ
authors.lastname: Cugier
authors.forename: R L
authors.initials: RL
authors.lastname: Bell
authors.forename: G W
authors.initials: GW
authors.lastname: Carter
authors.forename: K B
authors.initials: KB
authors.lastname: Glaser
mesh_terms: D000650:Amnion; D001483:Base Sequence; D015152:Blotting, Northern; D002460:Cell Line; D051546:Cyclooxygenase 2; D017931:DNA Primers; D015232:Dinoprostone / Q000032:analysis; D004790:Enzyme Induction; D005260:Female; D006801:Humans; D007249:Inflammation; D053590:Interleukin 1 Receptor Antagonist Protein; D007375:Interleukin-1 / Q000096:biosynthesis / Q000494:pharmacology*; D007527:Isoenzymes / Q000096:biosynthesis*; D007700:Kinetics; D008565:Membrane Proteins; D008954:Models, Biological; D008969:Molecular Sequence Data; D019319:Okadaic Acid / Q000494:pharmacology; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D011451:Prostaglandin-Endoperoxide Synthases / Q000096:biosynthesis*; D012333:RNA, Messenger; D011994:Recombinant Proteins / Q000494:pharmacology; D012795:Sialoglycoproteins / Q000494:pharmacology; D013755:Tetradecanoylphorbol Acetate; D014158:Transcription, Genetic / Q000187:drug effects*; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis* / Q000494:pharmacology*
chemical_list: D017931:DNA Primers; C507763:IL1RN protein, human; D053590:Interleukin 1 Receptor Antagonist Protein; D007375:Interleukin-1; D007527:Isoenzymes; D008565:Membrane Proteins; D012333:RNA, Messenger; D011994:Recombinant Proteins; D012795:Sialoglycoproteins; D014409:Tumor Necrosis Factor-alpha; D019319:Okadaic Acid; D051546:Cyclooxygenase 2; C496540:PTGS2 protein, human; D011451:Prostaglandin-Endoperoxide Synthases; D015232:Dinoprostone; D013755:Tetradecanoylphorbol Acetate