Possible induction of fatty acid cyclooxygenase in mouse osteoblastic cells (MC3T3-E1) by cAMP

Prostaglandin E2 (PGE2), a bone-resorption factor, was essentially the sole arachidonate metabolite in an osteoblastic cell line cloned from mouse calvaria (MC3T3-E1). When the cells were cultured in the presence of 2% newborn bovine serum, 1 microM epinephrine markedly stimulated PGE2 synthesis from endogenous arachidonic acid. The PGE2 synthesis commenced after a lag phase of 1-2 h, and reached a maximum at about 3 h after the addition of epinephrine. The effect of epinephrine was inhibited by propranolol, and epinephrine could be replaced by isoproterenol, suggesting beta-adrenergic stimulation of PGE2 production. A rapid increase in intracellular cAMP was observed upon the addition of epinephrine. When the intracellular cAMP level was raised using cholera toxin or forskolin, the PGE2 synthesis was also stimulated. The enhanced PGE2 synthesis was attributed to an increased level of cyclooxygenase, which was shown by immunoprecipitation of the enzyme using anti-cyclooxygenase antibody. Inhibitors of transcription and translation suppressed the epinephrine-dependent increase in cyclooxygenase activity. These findings suggest induction of cyclooxygenase involving cAMP via an as yet unclarified mechanism.