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.
Department of Obstetrics, Gynecology and Reproductive Science, University of Saskatchewan, Saskatoon, Canada.
M R
MR
Rodway
C L
CL
Swan
N K
NK
Crellin
C
C
Gillio-Meina
P J
PJ
Chedrese
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