cAMP regulates P450scc gene expression by a cycloheximide-insensitive mechanism in cultured mouse Leydig MA-10 cells.
Mouse MA-10 Leydig tumor cells synthesize and secrete progesterone in response to human chorionic gonadotropin, luteinizing hormone, and cAMP but may not synthesize androgens. Maximal doses of human chorionic gonadotropin, ovine luteinizing hormone, forskolin, or 8-bromoadenosine 3',5'-cyclic monophosphate, stimulated cytochrome P450scc mRNA accumulation 1.5- to 3-fold and progesterone secretion 10- to 100-fold in MA-10 cells. P450scc mRNA increased by 2 hr and was maximal by 8 hr; polymerase run-on experiments showed this was due to increased P450scc gene transcription. MA-10 cells are a hormonally homogeneous population, as all cells expressed P450scc mRNA and responded to cAMP equally. cAMP-stimulated accumulation of P450scc mRNA continued in the presence of cycloheximide. Gonadotropins stimulated testicular steroidogenesis by coordinate cAMP-induced increases in P450scc gene transcription, mRNA accumulation, and P450scc activity. We cloned rat P450c17 cDNA and showed it detected no P450c17 mRNA in control or cAMP-stimulated MA-10 cells by RNA transfer blots or RNase protection assays. Similarly, HPLC detected no 17 alpha-hydroxyprogesterone or testosterone synthesis in MA-10 cells. Thus MA-10 cells, unlike untransformed Leydig cells, do not express detectable amounts of P450c17 mRNA or P450c17 activity.
Mellon
S H
SH
Department of Obstetrics, Gynecology, and Reproductive Sciences, University of California-San Francisco 94143.
Vaisse
C
C
D015124:8-Bromo Cyclic Adenosine Monophosphate / Q000494:pharmacology*; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002786:Cholesterol Side-Chain Cleavage Enzyme / Q000235:genetics*; D006063:Chorionic Gonadotropin / Q000494:pharmacology; D005576:Colforsin / Q000494:pharmacology*; D000242:Cyclic AMP / Q000502:physiology*; D003513:Cycloheximide / Q000494:pharmacology*; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D005796:Genes / Q000187:drug effects; D007984:Leydig Cell Tumor; D007986:Luteinizing Hormone / Q000494:pharmacology; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000235:genetics / Q000302:isolation & purification; D014158:Transcription, Genetic / Q000187:drug effects
D006063:Chorionic Gonadotropin; D012333:RNA, Messenger; D005576:Colforsin; D015124:8-Bromo Cyclic Adenosine Monophosphate; D007986:Luteinizing Hormone; D003513:Cycloheximide; D000242:Cyclic AMP; D002786:Cholesterol Side-Chain Cleavage Enzyme