Document Detail

Selection and optimization of MCF-7 cell line for screening selective inhibitors of 11beta-hydroxysteroid dehydrogenase 2.
MedLine Citation:
PMID:  20629036     Owner:  NLM     Status:  MEDLINE    
An 11beta-hydroxysteroid dehydrogenase type 1 (11beta-HSD1) produces glucocorticoid (GC) from 11-keto metabolite, and its modulation has been suggested as a novel approach to treat metabolic diseases. In contrast, type 2 isozyme 11beta-HSD2 is involved in the inactivation of glucocorticoids (GCs), protecting the non-selective mineralocorticoid receptor (MR) from GCs in kidney. Therefore, when 11beta-HSD1 inhibitors are pursued to treat the metabolic syndrome, preferential selectivity of inhibitors for type 1 over type 2 isozyme is rather important than inhibitory potency. Primarily, to search for cell lines with 11beta-HSD2 activity, we investigated the expression profiles of enzymes or receptors relevant to GC metabolism in breast, colon, and bone-derived cell lines. We demonstrated that MCF-7 cells had high expression for 11beta-HSD2, but not for 11beta-HSD1 with its cognate receptor. Next, for the determination of enzyme activity indirectly, we adopted homogeneous time resolved fluorescence (HTRF) cortisol assay. Obviously, the feasibility of HTRF to cellular 11beta-HSD2 was corroborated by constructing inhibitory response to an 11b-HSD2 inhibitor glycyrrhetinic acid (GA). Taken together, MCF-7 that overexpresses type 2 but not type 1 enzyme is chosen for cellular 11beta-HSD2 assay, and our results show that a nonradioactive HTRF assay is applicable for type 2 as well as type 1 isozyme.
Chi Hyun Kim; Young Sik Cho
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Publication Detail:
Type:  Evaluation Studies; Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Cell biochemistry and function     Volume:  28     ISSN:  1099-0844     ISO Abbreviation:  Cell Biochem. Funct.     Publication Date:  2010 Aug 
Date Detail:
Created Date:  2010-08-30     Completed Date:  2010-12-13     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  8305874     Medline TA:  Cell Biochem Funct     Country:  England    
Other Details:
Languages:  eng     Pagination:  440-7     Citation Subset:  IM    
Division of Electron Microscopic Research, Korea Basic Science Institute, 113 Gwahangno, Yuseong-gu, Daejeon, South Korea.
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MeSH Terms
11-beta-Hydroxysteroid Dehydrogenase Type 2 / analysis,  antagonists & inhibitors*,  metabolism
Cell Line, Tumor
Drug Screening Assays, Antitumor / methods*
Enzyme Assays / methods
Enzyme Inhibitors / pharmacology*
Hydrocortisone / analysis,  metabolism
Reg. No./Substance:
0/Enzyme Inhibitors; 50-23-7/Hydrocortisone; EC Dehydrogenase Type 2

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