title: Regulation of mRNA expression of 3 beta-hydroxy-5-ene steroid dehydrogenase in porcine granulosa cells in culture: a role for the protein kinase-C pathway.
abstract: We studied the effect of the tumor-promoting phorbol ester phorbol 12-myristate 13-acetate (PMA), which activates protein kinase-C, on porcine granulosa cells in culture. PMA as well as cholera toxin, forskolin, and hCG increased cAMP accumulation. PMA further augmented the elevation in cAMP accumulation induced by cholera toxin, forskolin, and hCG. In the same cell culture model, hCG induced a time-dependent increase in the 3 beta-hydroxy-5-ene steroid dehydrogenase (3 beta HSD) mRNA levels with a maximal 3-fold stimulation obtained at 8-16 h of incubation with 1 IU hCG/ml. PMA inhibited the increase in 3 beta HSD mRNA levels induced by hCG in a dose-dependent manner. The phorbol ester also inhibited the increase in 3 beta HSD mRNA levels stimulated by LH as well as cholera toxin and forskolin and the cAMP analogs (Bu)2cAMP and 8-bromo-cAMP. Activation of protein kinase-C by mezerein similarly inhibited hCG stimulation of 3 beta HSD mRNA levels. The present data indicate that activation of the protein kinase-C pathway induces generation of cAMP, but causes a near-complete inhibition of the stimulatory effects of hCG, LH, forskolin, cholera toxin, and cAMP analogs on 3 beta HSD mRNA levels in porcine granulosa cells in culture.
authors.affiliation: Reproductive Biology Research Unit, University of Saskatchewan, Saskatoon, Canada.
authors.forename: P J
authors.initials: PJ
authors.lastname: Chedrese
authors.forename: D
authors.initials: D
authors.lastname: Zhang
authors.forename: V
authors.initials: V
authors.lastname: Luu The
authors.forename: F
authors.initials: F
authors.lastname: Labrie
authors.forename: A V
authors.initials: AV
authors.lastname: Juorio
authors.forename: B D
authors.initials: BD
authors.lastname: Murphy
mesh_terms: D015096:3-Hydroxysteroid Dehydrogenases / Q000235:genetics*; D015124:8-Bromo Cyclic Adenosine Monophosphate / Q000494:pharmacology; D000818:Animals; D003994:Bucladesine / Q000494:pharmacology; D002478:Cells, Cultured; D002772:Cholera Toxin / Q000494:pharmacology; D006063:Chorionic Gonadotropin / Q000494:pharmacology; D005576:Colforsin / Q000494:pharmacology; D000242:Cyclic AMP / Q000378:metabolism; D004224:Diterpenes; D004789:Enzyme Activation / Q000187:drug effects; D005260:Female; D006107:Granulosa Cells / Q000187:drug effects / Q000378:metabolism*; D007700:Kinetics; D007986:Luteinizing Hormone / Q000494:pharmacology; D011493:Protein Kinase C / Q000378:metabolism*; D012333:RNA, Messenger / Q000096:biosynthesis*; D013552:Swine; D013729:Terpenes / Q000494:pharmacology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
chemical_list: D006063:Chorionic Gonadotropin; D004224:Diterpenes; D012333:RNA, Messenger; D013729:Terpenes; D005576:Colforsin; D015124:8-Bromo Cyclic Adenosine Monophosphate; C011309:mezerein; D003994:Bucladesine; D007986:Luteinizing Hormone; D002772:Cholera Toxin; D000242:Cyclic AMP; D015096:3-Hydroxysteroid Dehydrogenases; C031954:delta(5)-3 beta-hydroxysteroid dehydrogenase; D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate