Document Detail

Are MDCK cells transfected with the human MRP2 gene a good model of the human intestinal mucosa?
MedLine Citation:
PMID:  12134946     Owner:  NLM     Status:  MEDLINE    
PURPOSE: To investigate whether Madin-Darby canine kidney cells transfected with the human MRP2 gene (MDCK-MRP2) are a good model of the human intestinal mucosa. METHODS: MRP2 expression in Caco-2 cells was compared with the expression of this efflux transporter in MDCK-wild type (MDCK-WT) and MDCK-MRP2 cells using Western blotting methods. The polarized efflux activities of MRP2 in the MDCK-MRP2, MDCK-WT. MDCK cells transfected with the human MDR1 gene (MDCK-MDR1), and Caco-2 cells were compared using vinblastine as a substrate. Apparent Michaelis-Menten constants (K(M), Vmax) for the efflux of vinblastine in Caco-2 and MDCK-MRP2 cells were determined in the presence of GF120918 (2 microM), which inhibits P-glycoprotein but does not affect MRP2. Apparent inhibitory constants (K(I)) of known substrates/inhibitors of MRP2 were determined by measuring their effects on the efflux of vinblastine in these cell lines. RESULTS: MDCK-MRP2 cells expressed higher levels of MRP2 than MDCK-WT and Caco-2 cells as measured by Western blotting technique. Two isoforms of MRP2 expressed in Caco-2 and MDCK cells migrated at molecular weights of 150 kD and 190 kD. In MDCK-MRP2 cells, the 150 kD isoform appeared to be overexpressed. MDCK-MRP2 cell monolayers exhibited higher polarized efflux of vinblastine than Caco-2 and MDCK-WT cell monolayers. K(M) values for vinblastine in Caco-2 and MDCK-MRP2 cells were determined to be 71.8+/-11.6 and 137.3+/-33.6 microM. respectively, and Vmax values were determined to be (0.54+/-0.03 and 2.45+/-0.31 pmolcm(-2)s(-1), respectively. Known substrates/inhibitors of MRP2 showed differences in their ability to inhibit vinblastine efflux in Caco-2 cells as compared to MDCK-MRP2 cells CONCLUSIONS: These data suggest that MDCK-MRP2 cells overexpress only the 150 kD isoform of MRP2. The 190 kD isoform, which was also found in Caco-2 cells and MDCK-WT cells, was present in MDCK-MRP2 cells but not over expressed. The apparent kinetics constants and affinities of some MRP2 substrates were different in Caco-2 cells and MDCK-MRP2 cells. These differences in substrate activity could result from differences in the relative expression levels of the MRP2 isoforms present in Caco-2 cells and MDCK-MRP2 cells and/or differences in the partitioning of substrates in these two cell membrane bilayers.
Fuxing Tang; Kazutoshi Horie; Ronald T Borchardt
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Publication Detail:
Type:  Comparative Study; Journal Article; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Pharmaceutical research     Volume:  19     ISSN:  0724-8741     ISO Abbreviation:  Pharm. Res.     Publication Date:  2002 Jun 
Date Detail:
Created Date:  2002-07-23     Completed Date:  2003-01-15     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  8406521     Medline TA:  Pharm Res     Country:  United States    
Other Details:
Languages:  eng     Pagination:  773-9     Citation Subset:  IM    
Department of Pharmaceutical Chemistry, The University of Kansas, Lawrence 66047, USA.
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MeSH Terms
Caco-2 Cells / metabolism
Cell Line / metabolism*,  physiology
Dose-Response Relationship, Drug
Drug Evaluation, Preclinical / methods
Intestinal Mucosa / metabolism*
Mitochondrial Proteins*
Ribosomal Proteins / genetics,  metabolism*
Saccharomyces cerevisiae Proteins*
Transfection / methods*
Vinblastine / pharmacokinetics
Grant Support
Reg. No./Substance:
0/MRP2 protein, S cerevisiae; 0/Mitochondrial Proteins; 0/Ribosomal Proteins; 0/Saccharomyces cerevisiae Proteins; 865-21-4/Vinblastine

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