Regulation of arachidonic acid release and prostaglandin E2 production in thymic epithelial cells by ATPgammaS and transforming growth factor-alpha.

The arachidonic acid metabolites produced by thymic epithelial cells play a pivotal role in thymocyte development. We have discovered that ATP and TGF-alpha regulate the arachidonic acid metabolism in TEA3A1 rat thymic epithelial cells by activating phospholipase A2 enzymatic activity. Our present study demonstrates that ATP and its nonhydrolyzable analog ATPgammaS stimulate both prostaglandin E2 production and Ca2+ influx in TEA3A1 cells. The stimulation of prostaglandin E2 production and Ca2+ influx by ATP is inhibited by pertussis toxin treatment, indicating that ATP mediates its effect by binding to a G-protein-coupled purinergic receptor. Treatment of cells with ATPgammaS and transforming growth factor-alpha results in a synergistic activation of phospholipase A2 and stimulation of prostaglandin E2 production. Results from experiments using an inhibitor of receptor-mediated Ca2+ influx indicate that the synergistic stimulation of prostaglandin E2 production by ATPgammaS and transforming growth factor-alpha requires ATPgammaS-mediated Ca2+ influx. The inhibitor of tyrosine kinase genistein also blocked both ATPgammaS- and ATPgammaS plus transforming growth factor-alpha-mediated stimulation of prostaglandin E2 production, indicating that the activation of phospholipase A2 may involve a protein tyrosine phosphorylation step.