Regulation of arachidonic acid metabolism in Madin-Darby canine kidney cells: stimulation of synthesis of the cyclooxygenase system by 12-O-tetradecanoyl-phorbol-13-acetate.

Stimulation of Madin-Darby canine kidney cells by 12-O-tetradecanoyl-phorbol-13-acetate (TPA) results in an increase in prostaglandin synthesis. We have measured the specific activity of the cyclooxygenase system in a cell-free assay and demonstrate that a fourfold induction occurs by 6 h poststimulation. This induction could be prevented by continuous treatment with either cycloheximide or actinomycin D. When cycloheximide was added to cells 6 h post-TPA stimulation we found only a 50% reduction in prostaglandin synthesis over the ensuing 6 h in vitro; this decrease in activity was not observed when actinomycin was used. On the other hand, cells stimulated with TPA for 6 h and subsequently treated with cycloheximide or actinomycin D ceased prostaglandin synthesis completely within 1 h. These results suggest that TPA stimulation of prostaglandin synthesis requires both transcriptional and translational events but that a factor(s) in addition to or in place of the cyclooxygenase might be crucial.