LIVER-TYPE 11-BETA-HYDROXYSTEROID DEHYDROGENASE CDNA ENCODES REDUCTASE BUT NOT DEHYDROGENASE-ACTIVITY IN INTACT MAMMALIAN COS-7 CELLS

S C LOW, K E CHAPMAN, C R W EDWARDS, J R SECKL

Research output: Contribution to journalArticlepeer-review

Abstract

11 beta-Hydroxysteroid dehydrogenase (11 beta-HSD) catalyses the metabolism of corticosterone to inert 11-dehydrocorticosterone, thus preventing glucocorticoid access to otherwise non-selective renal mineralocorticoid receptors (MRs), producing aldosterone selectivity in vivo. At least two isoforms of 11 beta-HSD exist. One isoform (11 beta-HSD1) has been purified from rat liver and an encoding cDNA cloned from a rat liver library. Transfection of rat 11 beta-HSD, cDNA into amphibian cells with a mineralocorticoid phenotype encodes 11 beta-reductase activity (activation of inert 11-dehydrocorticosterone) suggesting that 11 beta-HSD1 does not have the necessary properties to protect renal MRs from exposure to glucocorticoids. This function is likely to reside in a second 11 beta-HSD isoform. 11 beta-HSD1 is co-localized with glucocorticoid receptors (GRs) and may modulate glucocorticoid access to this receptor type. To examine the predominant direction of 11 beta-HSD1 activity in intact mammalian cells, and the possible role of 11 beta-HSD in regulating glucocorticoid access to GRs, we transfected rat 11 beta-HSD1 cDNA into a mammalian kidney-derived cell system (COS-7) which has little endogenous 11 beta-HSD activity or mRNA expression.

Homogenates of COS-7 cells transfected with increasing amounts of 11 beta-HSD cDNA exhibited a dose-related increase in 11 beta-dehydrogenase activity. In contrast, intact cells did not convert corticosterone to 11-dehydrocorticosterone over 24 h, but showed a clear dose-related 11 beta-reductase activity, apparent within 4 h of addition of 11-dehydrocorticosterone to the medium. To demonstrate that this reflected a change in functional intracellular glucocorticoids, COS-7 cells were co-transfected with an expression vector encoding GR and a glucocorticoid-inducible MMTV-LTR luciferase reporter construct, with or without 11 beta-HSD. Corticosterone induced MMTV-LTR luciferase expression in the presence or absence of 11 beta-HSD. 11-Dehydrocorticosterone was without activity in the absence of 11 beta-HSD, but induced MMTV-LTR luciferase activity in the presence of 11 beta-HSD. These results indicate that rat 11 beta-HSD1 can behave exclusively as a reductase in intact mammalian cells. Thus in some tissues in vivo, 11 beta-HSD1 may regulate ligand access to GRs by reactivating inert glucocorticoids.

Original languageEnglish
Pages (from-to)167-174
Number of pages8
JournalJournal of molecular endocrinology
Volume13
Issue number2
Publication statusPublished - Oct 1994

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