PMID- 9191983 OWN - NLM STAT- MEDLINE DCOM- 19970710 LR - 20190831 IS - 0960-0760 (Print) IS - 0960-0760 (Linking) VI - 60 IP - 3-4 DP - 1997 Feb TI - 11 beta-Hydroxysteroid dehydrogenase 1 activity and gene expression in human adipose stromal cells: effect on aromatase activity. PG - 247-53 AB - The biological activity of glucocorticoids in target tissues can be influenced by locally produced 11 beta-hydroxysteroid dehydrogenase (11 beta-HSD), the enzyme responsible for the interconversion of cortisol and its inactive metabolite cortisone. In human adipose stromal cells, glucocorticoids are potent stimulators of the conversion of androgens to estrogens (aromatase activity). The present study was designed to determine whether 11 beta-HSD activity was present in human adipose stromal cells, and if changes in the activity of this enzyme could influence aromatase activity. 11 beta-HSD activity was determined by a radiometric conversion assay in breast adipose tissue from six patients. It was found that both dehydrogenase (cortisol to cortisone) and reductase (cortisone to cortisol) activities were present in all six subjects, and the reductase activity was always predominant. Carbenoxolone (CBX), a potent inhibitor of 11 beta-HSD, added to the culture medium at 50 and 200 microM, resulted in 39 +/- 4% and 85 +/- 1% inhibition, respectively, of both reductase and dehydrogenase activity of 11 beta-HSD. To determine whether alterations in 11 beta-HSD could influence aromatase activity, the effect of CBX (200 microM) on cortisol- and cortisone-induced changes in the conversion of androstenedione to estrone was examined. CBX prevented the stimulatory effect of cortisone and minimally potentiated the stimulatory effect of cortisol on aromatase activity, reflecting an inhibition of the local activation of cortisone and the local metabolism of cortisol, respectively. In order to determine whether the product of the 11 beta-HSD 1 gene was responsible for the observed 11 beta-HSD activity, total RNA extracts from these cells were subjected to Northern blot analysis using human 11 beta-HSD 1 cDNA as the probe. A single 1.8 11 beta-HSD 1 transcript was detected, and its abundance was reduced by CBX. No 11 beta-HSD 2 mRNA was detected. The present results demonstrate that the 11 beta-HSD 1 gene is expressed and functional in human breast adipose stromal cells and that changes in 11 beta-HSD 1 activity result in alterations in aromatase activity. FAU - Yang, K AU - Yang K AD - Department of Obstetrics and Gynaecology and Physiology, University of Western Ontario, Lawson Research Institute, St Joseph's Health Centre, London, Canada. FAU - Khalil, M W AU - Khalil MW FAU - Strutt, B J AU - Strutt BJ FAU - Killinger, D W AU - Killinger DW LA - eng PT - Journal Article PL - England TA - J Steroid Biochem Mol Biol JT - The Journal of steroid biochemistry and molecular biology JID - 9015483 RN - 0 (Glucocorticoids) RN - 0 (Isoenzymes) RN - 2DI9HA706A (Estrone) RN - 409J2J96VR (Androstenedione) RN - EC 1.1.- (Hydroxysteroid Dehydrogenases) RN - EC 1.1.1.146 (11-beta-Hydroxysteroid Dehydrogenases) RN - EC 1.14.14.1 (Aromatase) RN - MM6384NG73 (Carbenoxolone) RN - V27W9254FZ (Cortisone) RN - WI4X0X7BPJ (Hydrocortisone) SB - IM MH - 11-beta-Hydroxysteroid Dehydrogenases MH - Adipose Tissue/cytology/*enzymology MH - Adolescent MH - Adult MH - Androstenedione/metabolism MH - Aromatase/*metabolism MH - Breast/cytology/*enzymology MH - Carbenoxolone/pharmacology MH - Cells, Cultured MH - Cortisone/pharmacology MH - Drug Interactions MH - Estrone/metabolism MH - Female MH - Gene Expression MH - Glucocorticoids/*metabolism MH - Humans MH - Hydrocortisone/pharmacology MH - Hydroxysteroid Dehydrogenases/*metabolism MH - Isoenzymes/*metabolism MH - Mammaplasty MH - Middle Aged MH - Stromal Cells/cytology EDAT- 1997/02/01 00:00 MHDA- 1997/02/01 00:01 CRDT- 1997/02/01 00:00 PHST- 1997/02/01 00:00 [pubmed] PHST- 1997/02/01 00:01 [medline] PHST- 1997/02/01 00:00 [entrez] AID - S0960-0760(96)00187-2 [pii] AID - 10.1016/s0960-0760(96)00187-2 [doi] PST - ppublish SO - J Steroid Biochem Mol Biol. 1997 Feb;60(3-4):247-53. doi: 10.1016/s0960-0760(96)00187-2.