Steroid regulation of progesterone synthesis in a stable porcine granulosa cell line: a role for progestins.

The objective of this investigation was to determine the effect of steroid hormones on the synthesis of progesterone in a stable porcine granulosa cell line, JC-410. We also examined the effect of steroid hormones on expression of the genes encoding the steroidogenic enzymes, cytochrome P450-cholesterol side chain cleavage (P450scc) and 3beta-hydroxy-5-ene steroid dehydrogenase (3beta-HSD). We observed that 48 h exposure of the JC-410 cells to estradiol-17beta (estradiol), androstenedione, 5alpha-dihydrotestosterone, levonorgestrel, and 5-cholesten-3beta, 25-diol (25-hydroxycholesterol) resulted in stimulation of progesterone synthesis. 25-Hydroxycholesterol augmented progesterone synthesis stimulated by estradiol, 5alpha-dihydrotestosterone, levonorgestrel and 8-bromoadenosine 3':5'-cyclic monophosphate (8-Br-cAMP). This increase in progesterone synthesis was additive with estradiol, 5alpha-dihydrotestosterone and levonorgestrel, and synergistic with 8-Br-cAMP. Cholera toxin, progesterone, levonorgestrel and androstenedione increased P450scc mRNA levels, whereas estradiol had no effect. Cholera toxin, progesterone and levonorgestrel increased 3beta-HSD mRNA levels, but estradiol and androstenedione had no effect. The results were interpreted to mean that estrogens, androgens and progestins regulate progesterone synthesis in the JC-410 cells. The effect of androgens appears to be mediated by stimulation of P450scc gene expression while progestins stimulate both P450scc and 3beta-HSD gene expression. Our results support the concept that progesterone is an autocrine regulator of its own synthesis in granulosa cells.