Metabolic and efflux properties of Caco-2 cells stably transfected with nuclear receptors

被引:30
作者
Korjamo, Timo
Monkkonen, Jukka
Uusitalo, Jouko
Turpeinen, Miia
Pelkonen, Olavi
Honkakoski, Paavo
机构
[1] Univ Kuopio, Dept Pharmaceut, FIN-70211 Kuopio, Finland
[2] Novamass Analyt Ltd, Oulu, Finland
[3] Univ Oulu, Dept Pharmacol & Toxicol, Oulu, Finland
基金
芬兰科学院;
关键词
Caco-2; CAR; drug absorption; metabolism; PXR;
D O I
10.1007/s11095-006-9068-4
中图分类号
O6 [化学];
学科分类号
0703 ;
摘要
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.
引用
收藏
页码:1991 / 2001
页数:11
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