title: Human chorionic gonadotropin decreases insulin-like growth factor-I gene transcription in rat Leydig cells.
abstract: Insulin-like growth factor-I (IGF-I) and hCG have synergistic effects on Leydig cell steroidogenesis in primary culture. In the present study, we investigated the effects of hCG on IGF-I gene transcription in Leydig cells. Purified Leydig cells (8-10 x 10(6) cells/100-mm dish) obtained from 50- to 65-day-old male Sprague-Dawley rats were cultured for 24 h. After medium change, hCG (0.1-10 ng/ml) or 8-bromo-cAMP (0.1 mM) was added, and cultures were continued for varying periods of time. In response to stimulation with hCG, there was a marked increase in the expression of cholesterol side-chain cleavage cytochrome P450 mRNA. In contrast, hCG caused time- and dose-dependent decrements in IGF-I mRNA levels. Both large [7.5-kilobase (kb)] and small (0.8- to 1.2-kb) species of IGF-I mRNAs were markedly decreased 6 h after treatment with hCG. hCG in a concentration of 0.1 ng/ml did not alter IGF-I mRNA levels. Higher concentrations of hCG (1 and 10 ng/ml) markedly decreased both 7.5- and 0.8- to 1.2-kb IGF-I mRNAs (80% and 56% reductions, respectively). 8-Bromo-cAMP (0.1 mM) also markedly reduced IGF-I mRNA levels. Finally, we evaluated the effects of hCG on the stability and transcription rates of IGF-I mRNA. We found that t1/2 of IGF-I mRNA for control Leydig cells was 3.86 h, which was not significantly different from that of hCG-treated cells (t1/2 = 3.41 h). This indicates that treatment with hCG did not change the stability of IGF-I mRNA. The average transcription rate per h for IGF-I mRNA decreased from 1 (for control cells) to 0.74 (for hCG-treated cells). The t1/2 values and rates of transcription for beta-actin were 7.39 and 7.16 h, and 1 and 0.94 for control and hCG-treated cells, respectively, showing that RNA stability and rates of transcription did not change significantly for the beta-actin transcript. In conclusion, we have unequivocally demonstrated that hCG decreases the expression and transcription of IGF-I mRNA in Leydig cells.
authors.affiliation: Medical Service, W. J. B. Dorn Veterans Hospital, Columbia, South Carolina 29201.
authors.forename: T
authors.initials: T
authors.lastname: Lin
authors.forename: D
authors.initials: D
authors.lastname: Wang
authors.forename: M L
authors.initials: ML
authors.lastname: Nagpal
authors.forename: W
authors.initials: W
authors.lastname: Chang
mesh_terms: D015124:8-Bromo Cyclic Adenosine Monophosphate / Q000494:pharmacology; D000199:Actins / Q000235:genetics; D000818:Animals; D006063:Chorionic Gonadotropin / Q000494:pharmacology*; D005786:Gene Expression Regulation / Q000187:drug effects*; D006801:Humans; D007334:Insulin-Like Growth Factor I / Q000235:genetics*; D007985:Leydig Cells / Q000187:drug effects / Q000378:metabolism*; D008297:Male; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D014158:Transcription, Genetic / Q000187:drug effects*
chemical_list: D000199:Actins; D006063:Chorionic Gonadotropin; D012333:RNA, Messenger; D015124:8-Bromo Cyclic Adenosine Monophosphate; D007334:Insulin-Like Growth Factor I