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Metabolic and Efflux Properties of Caco-2 Cells Stably Transfected with Nuclear Receptors

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Abstract

Purpose

To characterise in detail the patterns of expression and functional activities of CYP and efflux pump genes in Caco-2 cells stably transfected with human Pregnane X Receptor or murine Constitutive Androstane Receptor.

Materials and Methods

Cell lines transfected with nuclear receptors were treated with established ligands, and gene expression of CYP and efflux pump genes were quantified by qRT-PCR and Western blot. P-glycoprotein activity was assessed by measuring calcein-AM accumulation and bidirectional permeability coefficients of digoxin and quinidine. CYP activities were measured with both fluorescent and non-fluorescent substrates.

Results

hPXR and mCAR upregulated some CYP and efflux pump genes ligand dependently. P-glycoprotein level was increased, but CYP3A4 protein remained below the limit of detection. P-glycoprotein activity was markedly elevated in Caco/mCAR cells and more modestly in Caco/hPXR cells. CYP3A4 activity remained lower than that in vitamin D-treated Caco-2 cells.

Conclusions

Nuclear receptors can modulate the expression of metabolic genes in Caco-2 cells, but the overall level of metabolism could not be efficiently controlled. P-glycoprotein activity increased, but CYP activities remained very low.

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Abbreviations

A-B:

apical-to-basolateral

AQ:

absorptive quotient

B-A:

basolateral-to-apical

BCRP:

breast cancer resistance protein

BFC:

7-benzyloxy-4-(trifluoromethyl)-coumarin

CYP:

cytochrome P450

HFC:

4-(trifluoromethyl)-coumarin

HNF4:

hepatic nuclear factor 4

hPXR:

human pregnane X receptor

mCAR:

murine constitutive androstane receptor

MDR1:

multidrug resistance 1 gene (P-glycoprotein)

MRP2:

multidrug reistance associated Protein 2

SQ:

secretory quotient

VD3:

1a,25-dihydroxyvitamin D3

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Acknowledgments

This study was financially supported by the National Technology Agency of Finland (TEKES) and the Academy of Finland. We thank Dr. Kai Kaarniranta for the help with MDR1 Western blot, Ms. Paula Nyyssönen and Mr. Markku Taskinen for skilful technical assistance and Dr. Joann von Weissenberg for hel** to revise the language of the early drafts.

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Authors and Affiliations

  1. Department of Pharmaceutics, University of Kuopio, P.O.B. 1627 , FIN-70211, Kuopio, Finland

    Timo Korjamo, Jukka Mönkkönen & Paavo Honkakoski

  2. Novamass Analytical Ltd., Oulu, Finland

    Jouko Uusitalo

  3. Department of Pharmacology and toxicology, University of Oulu, Oulu, Finland

    Miia Turpeinen & Olavi Pelkonen

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  1. Timo Korjamo
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Correspondence to Timo Korjamo.

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Korjamo, T., Mönkkönen, J., Uusitalo, J. et al. Metabolic and Efflux Properties of Caco-2 Cells Stably Transfected with Nuclear Receptors. Pharm Res 23, 1991–2001 (2006). https://doi.org/10.1007/s11095-006-9068-4

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