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Growth Hormone (GH) and 17ß-Estradiol Regulation of the Expression of Mouse GH Receptor and GH-Binding Protein in Cultured Mouse Hepatocytes¹

Department of Biology, Sinsheimer Laboratories, University of California, Santa Cruz, California 95064

Address all correspondence and requests for reprints to: Dr. Frank Talamantes, Department of Biology, Sinsheimer Laboratories, University of California, Santa Cruz, California 95064. E-mail: prolactin{at}aol.com

In the present study, primary mouse hepatocytes from 8- to 10-week-old virgin female Swiss-Webster mice were perfused with collagenase (100 U/ml) using the two-step method. Isolated hepatocytes were plated in a rat tail type I collagen sandwich configuration to examine the regulation of GH receptor (GHR) and GH-binding protein (GHBP) expression by GH and 17ß-estradiol (E₂). After 48 h of initial plating, hepatocytes were divided into groups of five replicates and treated for 24 h with medium containing no hormones (controls), GH (100 ng/ml), E₂ (10^-9 M), E₂ (10^-9 M) plus GH (100 ng/ml), or E₂ plus GH and ICI 182–780 at different concentrations.

Treatment of hepatocytes with GH or E₂ alone did not have any effect on the cellular concentrations of GHBP and GHR. However, the combination of E₂ and GH up-regulated the cellular concentrations of GHBP and GHR 2- to 3-fold. GHBP and GHR messenger RNA concentrations were also up-regulated 2- to 3-fold. ICI 182–780, a competitive inhibitor of E₂ for the estrogen receptor (ER), at different concentrations inhibited the E₂ and GH-induced stimulation of GHBP and GHR. Furthermore, ER concentrations increased 5- to 7-fold in hepatocytes treated with E₂ and GH compared with those in untreated cells or cells treated with either E₂ or GH alone. In the present study we have shown that in cultured hepatocytes from virgin female mice, GH or E₂ alone did not affect the concentrations of GHBP and GHR. However, E₂ and GH together significantly up-regulated GHR and GHBP expression.

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