Tpa inhibits the synthesis of androgens and cortisol and enhances the synthesis of non-17α-hydroxylated steroids in cultured human adrenocortical cells

Previously, we showed that 12-0-tetracecanoyl phorbol 13-acetate (TPA), an activator of protein kinase C, is a raitogen for fetal human definitive zone adrenocortical cells in culture. In the present experiments, TPA inhibited forskolin-stimulated cortisol, dehydroepiandrosterone (DHEA), and dehydroepiandrosterone sulfate (DHEAS) synthesis, and enhanced forskolin-stimulated progesterone and corticosterone synthesis. These changes in the pattern of steroidogenesis were shown to result from changes in enzyme activities after forskolin treatment. TPA increased forskolin-stimulated 3β-phydroxysteroid dehydrogenase (3β-HSD) 2-fold, while depressing forskolin-stimulated 17c-hydroxylase to basal values. DHEA sulfotransferase increased 3-fold on transfer of human adrenocortical cells from serum-containing to defined, serumfree n:ediur_; TPA inhibited this increase. Experiments in which TPA was added at various times during the time course of forskolin stimulation of 17α-hydroxylase showed that TPA prevents the increase in the level of 17α-hydroxylase, and does not have a direct inhibitory effect on 17α-hydroxylase activity. TPA also inhibited stimulation of 17α-hydroxylase by cAMP analogs, indicating that the inhibition of 17α-hydroxylase by TPA is not due to an effect on adenylate cyclase. Previous experiments have shown that stimulation of intracellular cAMP is sufficient for androgen synthesis by the human adrenocorticel cell, under defined, serum-free conditions, and that its high rate of androgen synthesis likely results from the relative levels of 3p-HSD, 17α-hydroxylase, and DHEA sulfotransferase in the cell. Enzyme induction by cAMP results in increased production of both androgens and glucocorticoids, whereas activation of protein kinase C changes the balance of enzymes, resulting in increased non-17α.-hydroxylated steroid synthesis and decreased androgen and cortisol biosynthesis.