alexa Human chorionic gonadotropin decreases insulin-like growth factor-I gene transcription in rat Leydig cells.
Diabetes & Endocrinology

Diabetes & Endocrinology

Journal of Steroids & Hormonal Science

Author(s): Lin T, Wang D, Nagpal ML

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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.

This article was published in Endocrinology and referenced in Journal of Steroids & Hormonal Science

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