title: The effects of culture conditions on CYP3A4 and MDR1 mRNA induction by 1alpha,25-dihydroxyvitamin D(3) in human intestinal cell lines, Caco-2 and LS180.
abstract: To evaluate an in vitro model suitable for investigating intestinal first-pass drug metabolism, CYP3A4 and MDR1 mRNA induction by 1alpha,25-dihydroxyvitamin D(3) (VD3) was examined in two human intestinal cell lines, Caco-2 and LS180, under various culture conditions. CYP3A4 mRNA expression was induced by 100 nM VD3 at levels between 234-549 times above normal in Caco-2 cells for 2 weeks and by 74-200 times above normal in LS180 cells for 2 days. The CYP3A4 induction effect of 250 nM VD3 was similar to or slightly higher than that of 100 nM VD3 in both Caco-2 and LS180 cells. Also, CYP3A4 was induced in Caco-2 and LS180 cells when they were cultured on a polystyrene plate slightly less than when they were cultured on a porous membrane. The increase in fetal bovine serum (FBS) content in the culture medium resulted in little or only slight increase of CYP3A4 induction in both Caco-2 and LS180 cells. MDR1 mRNA expression was marginally increased by VD3 in LS180 cells, but not in Caco-2 cells, and neither increased FBS content nor use of a porous membrane significantly affected MDR1 induction in LS180 cells. The transepithelial electrical resistance of LS180 cells was almost zero, whereas that of Caco-2 cells was high and was marginally decreased by VD3. These findings indicate that Caco-2 cells cultured on a porous membrane with 100 nM VD3 for 2 weeks may be used as a model to investigate the intestinal absorption and first-pass metabolism of drugs, while LS180 cells may be utilized to elucidate the mechanisms which regulate intestinal CYP3A4 mRNA expression.
authors.affiliation: Graduate School of Pharmaceutical Sciences, Toyama Medical and Pharmaceutical University, Toyama, Japan.
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Aiba
authors.forename: Mitsuru
authors.initials: M
authors.lastname: Susa
authors.forename: Shiro
authors.initials: S
authors.lastname: Fukumori
authors.forename: Yukiya
authors.initials: Y
authors.lastname: Hashimoto
mesh_terms: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000096:biosynthesis* / Q000235:genetics; D018938:Caco-2 Cells; D002117:Calcitriol / Q000494:pharmacology*; D002460:Cell Line; D051544:Cytochrome P-450 CYP3A; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis* / Q000235:genetics; D004790:Enzyme Induction / Q000187:drug effects; D006801:Humans; D007422:Intestines / Q000187:drug effects / Q000201:enzymology; D023421:Models, Animal; D012333:RNA, Messenger / Q000096:biosynthesis; D013997:Time Factors
chemical_list: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D012333:RNA, Messenger; D003577:Cytochrome P-450 Enzyme System; D051544:Cytochrome P-450 CYP3A; C510163:CYP3A4 protein, human; D002117:Calcitriol