17-BETA-HYDROXYSTEROID DEHYDROGENASES IN HUMAN BONE-CELLS

Interconversion of estrogens by osteoblasts may play a role in regulat ing bone mass. As a first step toward exploring this possibility, we i nvestigated the expression and activity of 17 beta-hydroxysteroid dehy drogenases (17 beta-HSDs) in cultured human osteoblasts (HOB) and oste oblast-like osteosarcoma cells (MG63, TE85, and SaOS-2), Significant 1 7 beta-HSD activity was detected in cell-free extracts of all bone cel ls with oxidation of estradiol to estrone predominating over reduction . Reverse transcription-polymerase chain reaction (RT-PCR) experiments showed that the mRNA for 17 beta-HSD I was detectable only in MG63 ce lls, albeit at low levels, while 17 beta-HSD II was present in MG63, T E85, and HOB, but not SaOS-2, and 17 beta-HSD III was absent from each bone cell type, 17 beta-HSD Iv was the only isoform present in all bo ne cells analyzed. Further analysis of the expression of 17 beta-HSD I V in these bone cells by immunoblotting revealed both the full-length 83 kDa protein and the proteolytic 38 kDa form. The kinetic parameters for estradiol oxidation by purified recombinant 17 beta-HSD IV (K-m = 49.7 mu M, V-max = 79.4 nmol/minute/mg of protein) and its HSD-domain (K-m = 79.4 mu M, V-max = 476 nmol/minute/mg of protein) were signifi cantly higher than previously reported, but consistent with the values obtained with crude cell-free extracts of SaOS-2 cells (K-m = 98.8 mu M, V-max = 0.07 nmol/minute/mg of protein) which contain only 17 beta -HSD IV based on RT-PCR, These studies show that bone cells have the c apacity to interconvert circulating estrogens and suggest that bone ce ll 17 beta-HSDs serve primarily to attenuate the continuing actions of estradiol through conversion to its less potent form, estrone, under certain conditions.