11 beta-Hydroxysteroid dehydrogenase and tissue specificity of androgen action in human prostate cancer cell LNCaP.
Incubation of whole LNCaP cells in suspension with tritium labeled cortisol revealed two major and one minor radioactive product. Of the major products, one migrated with an Rf value identical to cortisol (Kendall's compound "F"), and the second migrated with an Rf value similar to nonradioactive cortisone (Kendall's compound "E"); the third minor product comigrated with 21-acetylated cortisol. The conversion of cortisol to cortisone was linear with respect to cell number, and conversion reached a plateau after 120 min of incubation at 37 degrees C. One half of the cortisol was converted to cortisone within 2 h of incubation at 37 degrees C. This conversion was nicotine amide dinucleotide (NAD) dependent. Low levels of transcription activation by cortisol were documented in LNCaP cells transfected with glucocorticoid and androgen responsive mouse mammary tumor virus-bacterial chloramphenicol acetyltransferase chimeric gene (MMTV-CAT). Hormone binding assay and transactivation analysis revealed the presence of a functional mineralocorticoid receptor in LNCaP cells. Treatment of transfectants with F in the presence of carbenoxolone, a potent inhibitor of 11 beta-hydroxysteroid dehydrogenase (11 beta-HSD), resulted in a two orders of magnitude increase in measurable CAT activity. The addition of the reduced form of nicotine amide dinucleotide (NADH) in the presence of 10(-7) M E stimulated measurable CAT activity in LNCaP cells. In conferring aldosterone specificity in mineralocorticoid target tissues, 11 beta-HSD may have an important role as "gate keeper" in allowing a specific androgen response in hormone responsive LNCaP prostate cancer cells.
M.R.C. Group in Molecular Endocrinology, Quebec, Canada.
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D042842:11-beta-Hydroxysteroid Dehydrogenases; D000107:Acetylation; D000450:Aldosterone / Q000378:metabolism; D000728:Androgens / Q000494:pharmacology*; D002229:Carbenoxolone / Q000494:pharmacology; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics / Q000378:metabolism; D003348:Cortisone / Q000378:metabolism; D006801:Humans; D006854:Hydrocortisone / Q000378:metabolism / Q000494:pharmacology; D006913:Hydroxysteroid Dehydrogenases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D008297:Male; D008324:Mammary Tumor Virus, Mouse / Q000235:genetics; D009243:NAD / Q000494:pharmacology; D011471:Prostatic Neoplasms / Q000201:enzymology* / Q000378:metabolism; D018161:Receptors, Mineralocorticoid / Q000378:metabolism; D014158:Transcription, Genetic / Q000187:drug effects; D014162:Transfection; D014316:Tritium; D014407:Tumor Cells, Cultured
D000728:Androgens; D018161:Receptors, Mineralocorticoid; D009243:NAD; D014316:Tritium; D000450:Aldosterone; D006913:Hydroxysteroid Dehydrogenases; D042842:11-beta-Hydroxysteroid Dehydrogenases; D015500:Chloramphenicol O-Acetyltransferase; D002229:Carbenoxolone; D003348:Cortisone; D006854:Hydrocortisone