Sn-1,2-diacylglycerols and phorbol esters stimulate the synthesis and release of human placental lactogen from placental cells: a role for protein kinase C.

Abstract

Activation of calcium-stimulated phospholipid-dependent protein kinase C (PKC) by diacylglycerols and phorbol esters has been shown to mediate the release of secretory proteins in several systems. To determine whether PKC activation is involved in regulation of the release of human placental lactogen (hPL) from the placenta, we examined the effects of various acylglycerols and phorbol esters on the release of hPL from cultured human trophoblast cells. Sn-1,2-dioctanoylglycerol (diC8) and phorbol-12-myristate-13-acetate (PMA), both of which stimulate placental protein kinase C activity, caused dose-dependent increases in hPL release over a 0.5-h period. The maximal amounts of hPL released in response to diC8 (300 microM) and PMA (10(-8) M) were 200-300% and 150-225% greater, respectively, than that released in response to diluent alone. Acylglycerols and phorbol esters, which are less potent stimulators of PKC activity in other systems, stimulated hPL release to a lesser extent than either diC8 or PMA. PKC-inactive acylglycerols and phorbol esters were without effect. After 0.5 h of exposure, diC8 (300 microM)- and PMA (10(-8) M)-exposed cells synthesized 257.5% and 250.3% more hPL than control cells. Cycloheximide at a dose (50 micrograms/ml) that inhibited the synthesis of trichloroacetic acid-precipitable [35S]methionyl placental proteins by more than 80% completely blocked the stimulatory effects of diC8 and PMA on hPL synthesis and release. Although diC8 and PMA stimulated the synthesis and release of hPL, these compounds had no effect on the release of hCG and did not cause the release of the cytosolic enzymes lactic dehydrogenase and alkaline phosphatase. The demonstration that acylglycerols and phorbol esters stimulate the synthesis and release of hPL strongly implicates protein kinase C activation in the mechanisms of hPL synthesis and release.