REGULATION OF HUMAN GROWTH-HORMONE RECEPTOR GENE-TRANSCRIPTION BY HUMAN GROWTH-HORMONE BINDING-PROTEIN

The hypothesis that growth hormone binding protein (GHBP) has an effec t on its own on the regulation of the GH-receptor/ GHBP transcription was tested. Three different forms of human GHBP (recombinant non-glyco sylated GHBP, recombinant glycosylated GHBP and GHBP purified and extr acted from serum) were added in different concentrations determined by LIFA [0 pmol/l; 50 pmol/l (low level), 200 pmol/l (average level) and 500 pmol/l (high level in circulation)] to a human hepatoma cell line (HuH7 cells) cultured in a serum free hormonally-defined medium. Foll owing the incubation with GHBP for 0, 1 and 2 h, GH-receptor expressio n was quantitatively assessed by using polymerase chain reaction ampli fication. Treatment with a GHBP concentration of 50 pmol/l resulted in a significant increase of GH-receptor mRNA molecules given as number of molecules x 10(6)/mu g total RNA. In contrast, the concentration of 500 pmol/l presented a significant decrease of GH-receptor mRNA molec ules, whereas 200 pmol/l GHBP produced a GH-receptor gene expression w hich was in between the values of the experiments with 50 and 500 pmol /l of GHBP added. Furthermore, the three different forms of human GHBP used provided similar data and, therefore; did not effect in any vari ation of GH-receptor expression. In addition, nuclear run-on experimen ts confirmed the changes in GH-receptor expression and cycloheximide ( 10 mu g/ml) did nor alter the transcription indicating that the up and down regulating effects of GHBP on the GH-receptor/GHBP gene transcri ption was dependent, at least partly, on pre-existing factors and does not require protein synthesis. In conclusion; we present data showing that GHBP on its own has an effect on GH-receptor gene expression. (C ) 1997 Elsevier Science Ireland Ltd.