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Biology of Reproduction 59, 1116-1123 (1998)
©Copyright 1998 Society for the Study of Reproduction, Inc.

Regulation of Luteinizing Hormone Receptor Gene Expression by Insulin-Like Growth Factor-I in an Immortalized Murine Leydig Tumor Cell Line (BLT-1)1

Fu-Ping Zhanga, Talal El-Hafnawya, , and Ilpo Huhtaniemi2,a

a Department of Physiology, University of Turku, FIN-20520 Turku, Finland

It is postulated that insulin-like growth factor-I (IGF-I), a 70-amino acid mitogenic polypeptide, regulates Leydig cell steroidogenesis. In the present study, we assessed the effect of IGF-I on LH receptor (LHR) gene expression in an immortalized murine Leydig tumor cell line (BLT-1). Culture of BLT-1 cells in the presence of IGF-I (0.1–100 ng/ml) for 24 or 48 h increased their [125I]iodo-hCG binding in a dose-dependent manner up to 275% of the control level. Northern hybridization analysis revealed four major transcripts of LHR mRNA in BLT-1 cells (6.9, 2.6, 1.7, and 1.2 kilobases), and treatment at 10–100 ng/ml of IGF-I increased steady-state levels of LHR mRNAs in coordinate fashion up to 2.2-fold. IGF-I (30 ng/ml) induced a time-dependent increase in [125I]hCG binding after a lag period of 2–6 h when studied up to 48 h, with a subsequent decrease. A similar response with steady increase up to 72 h was observed in total LHR mRNA. To elucidate the molecular mechanism of IGF-I action on LHR mRNA expression, we measured the transcription rate of the LHR gene by nuclear run-off assay and assessed transcript stability by the actinomycin D blocking method. The results showed that IGF-I treatment had no effect on the transcription rate of the LHR gene, whereas the half-life (t1/2) of LHR mRNA was significantly prolonged (IGF-I-treated cells, 30 ± 3.8 h; controls, 17 ± 2.5 h). Furthermore, IGF-I at 30 ng/ml and 100 ng/ml increased the expression of LHR promoter-driven luciferase and cytomegalovirus-promoter driven ß-galactosidase activities in BLT-1 cells; however, the former increased only marginally more than the latter. This suggests that the increase of LHR mRNA by IGF-I in Leydig cells is mainly due to increased mRNA stability.

1 This study was supported by grants from the Academy of Finland and the Sigrid Jusélius Foundation.

2 Correspondence: Ilpo Huhtaniemi, Department of Physiology, University of Turku, Kiinamyllynkatu 10, FIN-20520 Turku, Finland. FAX: 358 2 2502610; ilpo.huhtaniemi{at}utu.fi




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