Prostaglandin-induced changes in calcium uptake and cAMP production in osteoblast-like cells: role of protein kinase C.

Phorbol esters were used to evaluate the putative effect of protein kinase C (PKC) activation on prostaglandin E2 (PGE2)-induced increases in calcium uptake and cAMP production in the human osteoblastic osteosarcoma cell line, Saos-2. The cells were pretreated for 15 min with phorbol myristate acetate (PMA) followed by a 5 min incubation with PGE2. Calcium uptake was measured with 45Ca and cAMP by radioimmunoassay. A significant increase in calcium uptake was noted in the PGE-treated cells compared with controls and preincubation with the PMA caused a significant decrease in this response. Preincubation with PMA also inhibited the PGE2-induced increase in cAMP under identical conditions. The effect of PMA on the cAMP response was not influenced by the addition of a phosphodiesterase inhibitor. PMA had no effect on the basal levels of either calcium uptake or cAMP production. Likewise, the inactive phorbol esters, phorbol 12,13-didecanoate (PDD) and 4 alpha-phorbol 12-myristate, 13-acetate (4 alpha), had no effect on either basal levels of these parameters or on the PGE2-induced increases. These results suggest that PKC is involved in the down-regulation of PGE2-induced increases in calcium uptake and cAMP production in the Saos-2 osteoblastic cell line.