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.
Rodway
M R
MR
Department of Obstetrics, Gynecology and Reproductive Science, University of Saskatchewan, Saskatoon, Canada.
Swan
C L
CL
Crellin
N K
NK
Gillio-Meina
C
C
Chedrese
P J
PJ
D015096:3-Hydroxysteroid Dehydrogenases / Q000235:genetics; D000735:Androstenedione / Q000494:pharmacology; D000818:Animals; D002460:Cell Line; D002786:Cholesterol Side-Chain Cleavage Enzyme / Q000235:genetics; D003271:Contraceptive Agents, Female / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004958:Estradiol / Q000494:pharmacology; D005260:Female; D015971:Gene Expression Regulation, Enzymologic; D006107:Granulosa Cells / Q000187:drug effects* / Q000378:metabolism; D006888:Hydroxycholesterols / Q000494:pharmacology; D016912:Levonorgestrel / Q000494:pharmacology; D011374:Progesterone / Q000096:biosynthesis*; D011372:Progestins / Q000494:pharmacology* / Q000502:physiology; D013552:Swine
D003271:Contraceptive Agents, Female; D006888:Hydroxycholesterols; D011372:Progestins; D000735:Androstenedione; D011374:Progesterone; D004958:Estradiol; D016912:Levonorgestrel; C007997:25-hydroxycholesterol; D015096:3-Hydroxysteroid Dehydrogenases; D002786:Cholesterol Side-Chain Cleavage Enzyme