title: Genetically Modified Caco-2 Cells With Improved Cytochrome P450 Metabolic Capacity.
abstract: The human intestinal Caco-2 cell line has been extensively used as a model of small intestinal absorption but it lacks expression and function of cytochrome P450 enzymes, particularly CYP3A4 and CYP2C9, which are normally expressed in the intestinal epithelium. In order to increase the expression and activity of CYP isozymes in these cells, we created 2 novel Caco-2 sublines expressing chimeric constitutive androstane or pregnane X receptors and characterized these cells for their metabolic and absorption properties. In spite of elevated mRNA expression of transporters and differentiation markers, the permeation properties of the modified cell lines did not significantly differ from those of the wild-type cells. In contrast, the metabolic activity was increased beyond the currently used models. Specifically, CYP3A4 activity was increased up to 20-fold as compared to vitamin D treated wild-type Caco-2 cells.
authors: School of Pharmacy, University of Eastern Finland, FI-70211 Kuopio, Finland. Electronic address: jenni.kublbeck@uef.fi.
authors: Jenni
authors: J
authors: Küblbeck
authors: School of Pharmacy, University of Eastern Finland, FI-70211 Kuopio, Finland.
authors: Jenni J
authors: JJ
authors: Hakkarainen
authors: School of Pharmacy, University of Eastern Finland, FI-70211 Kuopio, Finland.
authors: Aleksanteri
authors: A
authors: Petsalo
authors: School of Pharmacy, University of Eastern Finland, FI-70211 Kuopio, Finland.
authors: Kati-Sisko
authors: KS
authors: Vellonen
authors: Admescope Ltd., Typpitie 1, FI-90620 Oulu, Finland.
authors: Ari
authors: A
authors: Tolonen
authors: Novamass Ltd., Viikinkaari 4, FI-00790 Helsinki, Finland.
authors: Petri
authors: P
authors: Reponen
authors: School of Pharmacy, University of Eastern Finland, FI-70211 Kuopio, Finland.
authors: Markus M
authors: MM
authors: Forsberg
authors: School of Pharmacy, University of Eastern Finland, FI-70211 Kuopio, Finland.
authors: Paavo
authors: P
authors: Honkakoski
mesh_terms: D018938:Caco-2 Cells; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics* / Q000378:metabolism*; D006801:Humans; D007408:Intestinal Absorption / Q000235:genetics / Q000502:physiology*; D053858:Metabolic Networks and Pathways / Q000235:genetics / Q000502:physiology*; D030781:Organisms, Genetically Modified; D014162:Transfection / Q000379:methods*
chemical_list: D003577:Cytochrome P-450 Enzyme System
keywords: Caco-2 cells; cytochrome P450; metabolism; nuclear receptors; permeability; transporters