title: Oxysterol activation of arachidonic acid release and prostaglandin E2 biosynthesis in NRK 49F cells is partially dependent on protein kinase C activity [corrected].
abstract: We previously demonstrated that the oxysterol potentiation of arachidonic acid release and prostaglandin biosynthesis induced by foetal calf serum activation of normal rat kidney (NRK) cells (fibroblastic clone 49F) was not related to a direct effect of oxysterols on cell free Ca2+ level. Since both Ca2+ variations and protein kinase C are involved in arachidonic acid release in some models, we looked for a possible modulation by protein kinase C in the oxysterol effect on arachidonic acid release. We show that when the phorbol ester 12-O-tetradecanoyl-phorbol-13-acetate (TPA), a protein kinase C activator, was added to the culture medium, the oxysterol effect on arachidonic acid release and prostaglandin synthesis clearly increased. Moreover, the effect of TPA was dose-dependent and TPA EC50 (4 x 10(-9) M) was unchanged in the presence of the oxysterol. Preincubation of cells with TPA for 24 h prevented the arachidonic acid release induced by TPA alone, whereas the oxysterol effect was decreased but not abolished. In the absence of serum, TPA and ionomycin added together induced the same noticeable (arachidonic acid) release and PGE2 synthesis as serum alone. Nevertheless, the potentiating effect of cholest-5-ene-3 beta, 25-diol was much higher when serum itself was used to activate NRK cells than it was in the present serum-mimicking experimental conditions. Thus, the presence of growth factors is probably required to obtain a full oxysterol effect. We conclude that the oxysterol effect was synergistic with, but not fully dependent on, protein kinase C and Ca2+ ion fluxes, therefore oxysterols could affect earlier events triggered by serum growth factor binding to their cell membrane receptors.
authors.affiliation: INSERM U.58, Montepellier, France.
authors.forename: Z
authors.initials: Z
authors.lastname: Lahoua
authors.forename: H
authors.initials: H
authors.lastname: Vial
authors.forename: F
authors.initials: F
authors.lastname: Michel
authors.forename: A
authors.initials: A
authors.lastname: Crastes de Paulet
authors.forename: M E
authors.initials: ME
authors.lastname: Astruc
mesh_terms: D000818:Animals; D016718:Arachidonic Acid / Q000378:metabolism*; D001769:Blood; D002117:Calcitriol / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism; D002460:Cell Line; D015232:Dinoprostone / Q000096:biosynthesis*; D006888:Hydroxycholesterols / Q000494:pharmacology*; D015759:Ionomycin / Q000494:pharmacology; D007653:Ketocholesterols / Q000494:pharmacology*; D011493:Protein Kinase C / Q000378:metabolism*; D051381:Rats; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
chemical_list: D006888:Hydroxycholesterols; D007653:Ketocholesterols; D016718:Arachidonic Acid; D015759:Ionomycin; D011493:Protein Kinase C; D002117:Calcitriol; D015232:Dinoprostone; D013755:Tetradecanoylphorbol Acetate; C003001:7-ketocholesterol; D002118:Calcium