Influence of drugs and nutrients on transporter gene expression levels in Caco-2 and LS180 intestinal epithelial cell lines.
OBJECTIVE
The aim of this study was to investigate the influences of various drugs and nutrients on the expression levels of intestinal drug transporters PEPT1, MDR1, MRP2 and MRP3, and drug-metabolizing enzyme CYP3A4.


METHODS
Quantitative reverse transcriptase polymerase chain reaction was used to quantitate transporter and CYP3A4 mRNAs. Western blotting was used to determine protein levels of P-gp. Transport studies of P-gp were performed using cultured Caco-2 cell monolayers.


RESULTS
The expression of MDR1 mRNA was increased by all-trans retinoic acid and in glucose-depleted medium, whereas little change of MRP2 and MRP3 mRNA was observed in Caco-2 cells. Substrates and inducers of P-gp or CYP3A4 tended to produce parallel changes in the expression of MDR1 and CYP3A4 mRNA in LS180 cells, whereas in Caco-2 cells no such coordinate response was observed, possibly due to the absence of the expression of steroid xenobiotic receptor (SXR) in this cell line.


CONCLUSIONS
Several drugs and nutrients were found to affect transporter gene expression level in two human intestinal epithelial cell lines. Since SXR is involved in some expression-regulatory processes, and Caco-2 cells lack SXR, LS180 cells as well as Caco-2 cells should be used for the study of the regulation of intestinal transporters.
Li
Qing
Q
Department of Pharmaceutical Biology, Faculty of Pharmaceutical Sciences, Kanazawa University, 13-1 Takara-machi, Kanazawa, Ishikawa 920-0934, Japan.
Sai
Yoshimichi
Y
Kato
Yukio
Y
Tamai
Ikumi
I
Tsuji
Akira
A
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000096:biosynthesis / Q000235:genetics; D001692:Biological Transport; D015153:Blotting, Western; D002352:Carrier Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D003470:Culture Media; D051544:Cytochrome P-450 CYP3A; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis / Q000235:genetics; D015870:Gene Expression / Q000187:drug effects*; D006801:Humans; D007413:Intestinal Mucosa / Q000166:cytology / Q000378:metabolism*; D026901:Membrane Transport Proteins / Q000096:biosynthesis / Q000235:genetics; D000089762:Multidrug Resistance-Associated Protein 2; D027425:ATP-Binding Cassette, Sub-Family C Proteins / Q000096:biosynthesis / Q000235:genetics; D000074011:Peptide Transporter 1; D004364:Pharmaceutical Preparations / Q000378:metabolism*; D012333:RNA, Messenger / Q000096:biosynthesis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D027981:Symporters
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D002352:Carrier Proteins; D003470:Culture Media; D051544:Cytochrome P-450 CYP3A; D003577:Cytochrome P-450 Enzyme System; D026901:Membrane Transport Proteins; D000089762:Multidrug Resistance-Associated Protein 2; D027425:ATP-Binding Cassette, Sub-Family C Proteins; D000074011:Peptide Transporter 1; D004364:Pharmaceutical Preparations; D012333:RNA, Messenger; D027981:Symporters; C000715087:ABCC2 protein, human; C082186:SLC15A1 protein, human; C114462:multidrug resistance-associated protein 3; C104464:CYP3A protein, human; C510163:CYP3A4 protein, human; C421109:multidrug resistance-associated protein 1