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In thyroid follicular cells, the formation of cyclooxygenase-derived prostaglandin E2 (PGE2) is stimulated by Ca2+ influx through voltage-sensitive Ca2+ channels (VSCC) or by activation of phospholipase C (PLC). In the present study, the role of protein kinase C (PKC) in the PGE2 formation in thyroid C cells was evaluated by incubating the cells with the PKC activator PMA and by administering the PKC antagonists H7 and staurosporine. Incubation of thyrocytes for 60 min with PMA increased the formation of PGE2 by two- to threefold. PGE2 formation was inhibited by H7 and staurosporine. Immunoblot analysis revealed that PMA increased the level of PGE2 cyclooxygenase (COX)-2 by 1.6-fold. H7 and staurosporine inhibited this increase. The active enzyme was 2.1-fold more sensitive to the inhibitor than the inactive enzyme. PMA also increased the intracellular Ca2+ concentration ([Ca2+]i) from 25.9 +/- 1.9 to 48.6 +/- 2.2 nmol/L. Treatment of thyrocytes with H7 before incubation with PMA reduced [Ca2+]i from 25.9 +/- 1.9 to 20.9 +/- 1.8 nmol/L, and staurosporine completely abolished it. Thus, the PGE2 formation may be stimulated by PMA through a PKC-dependent

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