Metabolic and Efflux Properties of Caco-2 Cells Stably Transfected with Nuclear Receptors
| Autor: | Jouko Uusitalo, Jukka Mönkkönen, Olavi Pelkonen, Paavo Honkakoski, Miia Turpeinen, Timo Korjamo |
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| Rok vydání: | 2006 |
| Předmět: |
ATP Binding Cassette Transporter
Subfamily B Blotting Western Receptors Cytoplasmic and Nuclear Pharmaceutical Science Biology Ligands Transfection digestive system Permeability Cytochrome P-450 Enzyme System Gene expression Constitutive androstane receptor Cytochrome P-450 CYP3A Humans Pharmacology (medical) ATP Binding Cassette Transporter Subfamily B Member 1 RNA Messenger Receptor Pharmacology Pregnane X receptor Reverse Transcriptase Polymerase Chain Reaction Organic Chemistry Fluoresceins Molecular biology Nuclear receptor Biochemistry Cell culture Molecular Medicine Efflux Caco-2 Cells Biotechnology |
| Zdroj: | Pharmaceutical Research. 23:1991-2001 |
| ISSN: | 1573-904X 0724-8741 |
| Popis: | 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. 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. 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. 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. |
| Databáze: | OpenAIRE |
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