Regulation of cytochrome P450 cholesterol side-chain cleavage, 3 β-hydroxysteroid dehydrogenase/Δ5-Δ4 isomerase type 1 and estradiol-17 β-hydroxysteroid dehydrogenase mRNA levels by calcium in human choriocarcinoma JEG-3 cells

Abstract

In human placenta the cytochrome P450 side-chain cleavage (P450scc) and 3 β-hydroxysteroid dehydrogenase type 1 (3 β-HSD-1) convert cholesterol and pregnenolone producing progesterone, whereas 17 β-hydroxysteroid dehydrogenase type 1 (17 β-HSD-1) mediates the interconversion of estrone and estradiol. We have examined the effects of calcium on phorbol ester- and cAMP-induced P450scc, 3 β-HSD-1 and 17 β-HSD-1 mRNAs in human JEG-3 cells. A23187 increased in a dose-dependent fashion in the 1.3 kb 17 β-HSD-1 mRNA whereas a weaker increase followed by a gradual depletion effect of A23187 was observed on 3 β-HSD-1 mRNA. No significant effect of A23187 on P450scc mRNA was observed. Using 0.50 μM of A23187 the induction of 3 β-HSD-1 and 17 β-HSD-1 mRNAs was maximum within about 6 h whereas P450scc mRNA levels stayed unaffected throughout the time-course period. The action of A23187 was synergistic on cAMP-stimulated 17 β-HSD-1 mRNA levels, while in a dose-dependent manner A23187 progressively depleted 3 β-HSD-1 and P450scc mRNA abundance probably by activation of a calcium-/calmodulin-dependent phosphodiesterase. On the phorbol 12-myristate, 13-acetate (PMA)-stimulated 3 β-HSD-1, 17 β-HSD-1 and P450scc mRNA levels only the lowest concentration of A23187 potentialized the PMA effect on the 17 β-HSD-1 mRNA levels. Using thapsigargin (TG), a cell-permeable sesquiterpene lactone that releases calcium by inhibiting sarco/endoplasmic reticular calcium-ATPase, our data indicated the presence in JEG-3 cells of TG-sensitive and TG-insensitive calcium-ATPases regulating 3 β-HSD-1 and 17 β-HSD-1 mRNA levels. These results emphasized the complexity of calcium contribution with the protein kinase A and C pathways in the regulation of P450scc, 3 β-HSD-1 and 17 β-HSD-1 mRNA levels. In addition, the different sensitivity of these genes to calcium suggest they could be activated by different subclasses of PKCs.