Regulation of prostaglandin E2 synthesis in human amnion by protein kinase C.
A role for protein kinase C (PKC) in mediation of prostaglandin E2 synthesis in human amnion cells has been suggested. We have investigated the specificity of the stimulation of PGE2 synthesis by phorbol esters and employed putative PKC inhibitors to demonstrate the specificity of PKC stimulation. The three phorbol esters, tetradecanoyl phorbol-13-acetate, phorbol-12,13-dibutyrate and phorbol-12,13-didecanoate gave concentration-dependent (10(-10)-10(-7)M) increases in PGE2 synthesis when added to amnion cells in monolayer, however, no effect was seen with the structurally similar phorbols phorbol-12-13-diacetate, 4 alpha phorbol-12-13-didecanoate or phorbol base. The stimulatory effect of TPA (10(-8)M) on amnion PGE2 synthesis could be prevented by coincubation with the putative protein kinase C inhibitors 1-(5-isoquinoline sulphonyl) piperazine, 1-0-octadecyl-2-0-methyl-rac-glycero-3-phosphocholine, sphingosine and chlopromazine at concentrations of 10(-6)-10(-4)M. Addition of the transcription inhibitor actinomycin D at 10(-6)-10(-5)M prevented TPA (10(-8)M)-induced PGE2 synthesis. However, paradoxically, a further increase in PGE2 synthesis was seen when 10(-9)-10(-7)M actinomycin D was added together with TPA. The phospholipase A2 inhibitor quinacrine was able to prevent the TPA-induced increase in PGE2 synthesis even in the presence of exogenous arachidonic acid suggesting that phospholipase A2 may be a target for PKC action.
Sander
J
J
Department of Obstetrics and Gynecology, University of Cincinnati, College of Medicine, Ohio 45267-0526.
Myatt
L
L
D000650:Amnion / Q000378:metabolism*; D016718:Arachidonic Acid; D001095:Arachidonic Acids / Q000378:metabolism; D002478:Cells, Cultured; D015232:Dinoprostone / Q000096:biosynthesis*; D004305:Dose-Response Relationship, Drug; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D008563:Membrane Lipids / Q000378:metabolism; D010703:Phorbol Esters / Q000494:pharmacology*; D010741:Phospholipases A / Q000037:antagonists & inhibitors; D054467:Phospholipases A2; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000502:physiology*; D011796:Quinacrine / Q000494:pharmacology; D013268:Stimulation, Chemical; D013329:Structure-Activity Relationship
D001095:Arachidonic Acids; D008563:Membrane Lipids; D010703:Phorbol Esters; D016718:Arachidonic Acid; D011493:Protein Kinase C; D010741:Phospholipases A; D054467:Phospholipases A2; D011796:Quinacrine; D015232:Dinoprostone