Stimulation of cultured amnion cell prostaglandin endoperoxide H synthase activity by glucocorticoids and phorbol ester.
OBJECTIVE
The effects of a phorbol ester activator of protein kinase C and the glucocorticoids cortisol and dexamethasone on the enzyme activity of prostaglandin H synthase (cyclooxygenase) in confluent cultures of human amnion epithelial cells were tested.


METHODS
Amnion epithelial cells from spontaneously delivered placentas at term were isolated and grown in culture until confluent. The activity of the enzyme prostaglandin H synthase was determined in these cells with a well-characterized enzyme assay monitoring the conversion of arachidonic acid to prostaglandin E2. The Michaelis-Menten constant and maximum velocity were determined from the substrate velocity data by means of Lineweaver-Burk plots.


RESULTS
Amnion cells lost most of their prostaglandin H synthase activity within 2 days of culturing. This activity could be restored when cells were treated with 12-O-tetradecanoyl phorbol-13-acetate, a phorbol ester activator of protein kinase C, or with the glucocorticoids cortisol and dexamethasone. Epidermal growth factor also increased the specific activity, whereas 17 beta-estradiol had no effect on the specific activity of the enzyme. There was a direct correlation between the specific activity of prostaglandin H synthase and the output of prostaglandin E2 by cells treated with these agonists.


CONCLUSIONS
Our data support the view that increases in prostaglandin H synthase specific activity in intrauterine tissues can be caused by stimulation by specific agonists, and this in turn is responsible for enhanced prostaglandin output by these tissues.
Smieja
Z
Z
University of Alberta Perinatal Research Centre, Edmonton, Canada.
Zakar
T
T
Olson
D M
DM
D000650:Amnion / Q000187:drug effects / Q000201:enzymology*; D000704:Analysis of Variance; D016718:Arachidonic Acid / Q000378:metabolism; D002478:Cells, Cultured; D003907:Dexamethasone / Q000494:pharmacology*; D015232:Dinoprostone / Q000096:biosynthesis / Q000302:isolation & purification; D004305:Dose-Response Relationship, Drug; D006801:Humans; D006854:Hydrocortisone / Q000494:pharmacology*; D011451:Prostaglandin-Endoperoxide Synthases / Q000096:biosynthesis*; D011493:Protein Kinase C / Q000502:physiology; D012044:Regression Analysis; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*
D016718:Arachidonic Acid; D003907:Dexamethasone; D011451:Prostaglandin-Endoperoxide Synthases; D011493:Protein Kinase C; D015232:Dinoprostone; D013755:Tetradecanoylphorbol Acetate; D006854:Hydrocortisone