BACKGROUND AND PURPOSE: The organic cation transporter 1 (OCT1) transports cationic drugs into hepatocytes. The high hepatic expression of OCT1 is controlled by the HNF4α and USF transcription factors. Pregnane X receptor (PXR) mediates induction of the principal xenobiotic metabolizing enzymes and transporters in the liver. Here, we have assessed the down-regulation of OCT1 expression by PXR activation. EXPERIMENTAL APPROACH: We used primary human hepatocytes and related cell lines to measure OCT1 expression and activity, by assaying MPP(+) accumulation. Western blotting, qRT-PCR, the OCT1 promoter gene reporter constructs and chromatin immunoprecipitation assays were also used. KEY RESULTS: OCT1 mRNA in human hepatocytes was down-regulated along with reduced [(3) H]MPP(+) accumulation in differentiated HepaRG cells after treatment with rifampicin. Rifampicin and hyperforin as well as the constitutively active PXR mutant T248D suppressed activity of the 1.8 kb OCT1 promoter construct in gene reporter assays. Silencing of both PXR and HNF4α in HepaRG cells blocked the PXR ligand-mediated down-regulation of OCT1 expression. The mutation of HNF4α and USF1 (E-box) responsive elements reversed the PXR-mediated inhibition in gene reporter assays. Chromatin immunoprecipitation assays indicated that PXR activation sequestrates the SRC-1 coactivator from the HNF4α response element and E-box of the OCT1 promoter. Consistent with these findings, exogenous overexpression of the SRC-1, but not the PGC1α coactivator, relieved the PXR-mediated repression of OCT1 transactivation. CONCLUSIONS AND IMPLICATIONS: PXR ligands reduced the HNF4α-mediated and USF-mediated transactivation of OCT1 gene expression by competing for SRC-1 and decreased delivery of a model OCT1 substrate into hepatocytes.

Department of Pharmacology and Toxicology Faculty of Pharmacy in Hradec Kralove Charles University Prague Hradec Kralove Czech Republic

INSERM U1183 Institute for Regenerative Medicine and Biotherapy Montpellier France

Université de Montpellier Montpellier France

Zobrazit více v PubMed

Alexander SP, Kelly E, Marrion N, Peters JA, Benson HE, Faccenda E et al. (2015a). The Concise Guide to PHARMACOLOGY 2015/16: Transporters. Br J Pharmacol 172: 6110–6202. PubMed PMC

Alexander SP, Cidlowski JA, Kelly E, Marrion N, Peters JA, Benson HE et al. (2015b). The Concise Guide to PHARMACOLOGY 2015/16: Nuclear hormone receptors. Br J Pharmacol 172: 5956–5978. PubMed PMC

Austin G, Holcroft A, Rinne N, Wang L, Clark RE (2015). Evidence that the pregnane X and retinoid receptors PXR, RAR and RXR may regulate transcription of the transporter hOCT1 in chronic myeloid leukaemia cells. Eur J Haematol 94: 74–78. PubMed

Badolo L, Bundgaard C, Garmer M, Jensen B (2013). The role of hepatic transport and metabolism in the interactions between pravastatin or repaglinide and two rOatp inhibitors in rats. Eur J Pharm Sci 49: 767–772. PubMed

Becker ML, Visser LE, van Schaik RH, Hofman A, Uitterlinden AG, Stricker BH (2009). Genetic variation in the organic cation transporter 1 is associated with metformin response in patients with diabetes mellitus. Pharmacogenomics J 9: 242–247. PubMed

Boxberger KH, Hagenbuch B, Lampe JN (2014). Common drugs inhibit human organic cation transporter 1 (OCT1)‐mediated neurotransmitter uptake. Drug Metab Dispos 42: 990–995. PubMed PMC

Cahill MA, Ernst WH, Janknecht R, Nordheim A (1994). Regulatory squelching. FEBS Lett 344: 105–108. PubMed

Curtis MJ, Bond RA, Spina D, Ahluwalia A, Alexander SP, Giembycz MA et al. (2015). Experimental design and analysis and their reporting: new guidance for publication in BJP. Br J Pharmacol 172: 3461–3471. PubMed PMC

Doricakova A, Novotna A, Vrzal R, Pavek P, Dvorak Z (2013). The role of residues T248, Y249 and T422 in the function of human pregnane X receptor. Arch Toxicol 87: 291–301. PubMed

Gripon P, Rumin S, Urban S, Le Seyec J, Glaise D, Cannie I et al. (2002). Infection of a human hepatoma cell line by hepatitis B virus. Proc Natl Acad Sci U S A 99: 15655–15660. PubMed PMC

Hilgendorf C, Ahlin G, Seithel A, Artursson P, Ungell AL, Karlsson J (2007). Expression of thirty‐six drug transporter genes in human intestine, liver, kidney, and organotypic cell lines. Drug Metab Dispos 35: 1333–1340. PubMed

Huang S, Li X, Yusufzai TM, Qiu Y, Felsenfeld G (2007). USF1 recruits histone modification complexes and is critical for maintenance of a chromatin barrier. Mol Cell Biol 27: 7991–8002. PubMed PMC

Chen Y, Tang Y, Guo C, Wang J, Boral D, Nie D (2012). Nuclear receptors in the multidrug resistance through the regulation of drug‐metabolizing enzymes and drug transporters. Biochem Pharmacol 83: 1112–1126. PubMed PMC

Cho SK, Yoon JS, Lee MG, Lee DH, Lim LA, Park K et al. (2011). Rifampin enhances the glucose‐lowering effect of metformin and increases OCT1 mRNA levels in healthy participants. Clin Pharmacol Ther 89: 416–421. PubMed

Jigorel E, Le Vee M, Boursier‐Neyret C, Parmentier Y, Fardel O (2006). Differential regulation of sinusoidal and canalicular hepatic drug transporter expression by xenobiotics activating drug‐sensing receptors in primary human hepatocytes. Drug Metab Dispos 34: 1756–1763. PubMed

Jonker JW, Schinkel AH (2004). Pharmacological and physiological functions of the polyspecific organic cation transporters: OCT1, 2, and 3 (SLC22A1‐3). J Pharmacol Exp Ther 308: 2–9. PubMed

Jonker JW, Wagenaar E, Van Eijl S, Schinkel AH (2003). Deficiency in the organic cation transporters 1 and 2 (Oct1/Oct2 [Slc22a1/Slc22a2]) in mice abolishes renal secretion of organic cations. Mol Cell Biol 23: 7902–7908. PubMed PMC

Kajiwara M, Terada T, Asaka J, Aoki M, Katsura T, Ikai I et al. (2008). Regulation of basal core promoter activity of human organic cation transporter 1 (OCT1/SLC22A1). Am J Physiol Gastrointest Liver Physiol 295: G1211–G1216. PubMed

Kamiyama Y, Matsubara T, Yoshinari K, Nagata K, Kamimura H, Yamazoe Y (2007). Role of human hepatocyte nuclear factor 4alpha in the expression of drug‐metabolizing enzymes and transporters in human hepatocytes assessed by use of small interfering RNA. Drug Metab Pharmacokinet 22: 287–298. PubMed

Kliewer SA, Goodwin B, Willson TM (2002). The nuclear pregnane X receptor: a key regulator of xenobiotic metabolism. Endocr Rev 23: 687–702. PubMed

Koepsell H, Lips K, Volk C (2007). Polyspecific organic cation transporters: structure, function, physiological roles, and biopharmaceutical implications. Pharm Res 24: 1227–1251. PubMed

Le Vee M, Jigorel E, Glaise D, Gripon P, Guguen‐Guillouzo C, Fardel O (2006). Functional expression of sinusoidal and canalicular hepatic drug transporters in the differentiated human hepatoma HepaRG cell line. Eur J Pharm Sci 28: 109–117. PubMed

Mandikova J, Volkova M, Pavek P, Cesnek M, Janeba Z, Kubicek V et al. (2013). Interactions with selected drug renal transporters and transporter‐mediated cytotoxicity in antiviral agents from the group of acyclic nucleoside phosphonates. Toxicology 311: 135–146. PubMed

Miao J, Fang S, Bae Y, Kemper JK (2006). Functional inhibitory cross‐talk between constitutive androstane receptor and hepatic nuclear factor‐4 in hepatic lipid/glucose metabolism is mediated by competition for binding to the DR1 motif and to the common coactivators, GRIP‐1 and PGC‐1alpha. J Biol Chem 281: 14537–14546. PubMed

Min G, Kim H, Bae Y, Petz L, Kemper JK (2002). Inhibitory cross‐talk between estrogen receptor (ER) and constitutively activated androstane receptor (CAR). CAR inhibits ER‐mediated signaling pathway by squelching p160 coactivators. J Biol Chem 277: 34626–34633. PubMed

Nies AT, Koepsell H, Winter S, Burk O, Klein K, Kerb R et al. (2009). Expression of organic cation transporters OCT1 (SLC22A1) and OCT3 (SLC22A3) is affected by genetic factors and cholestasis in human liver. Hepatology 50: 1227–1240. PubMed

Pawson AJ, Sharman JL, Benson HE, Faccenda E, Alexander SP, Buneman OP et al. (2014). The IUPHAR/BPS Guide to PHARMACOLOGY: an expert‐driven knowledgebase of drug targets and their ligands. Nucleic Acids Res 42: D1098–D1106. PubMed PMC

Rulcova A, Krausova L, Smutny T, Vrzal R, Dvorak Z, Jover R et al. (2013). Glucocorticoid receptor regulates organic cation transporter 1 (OCT1, SLC22A1) expression via HNF4alpha upregulation in primary human hepatocytes. Pharmacol Rep 65: 1322–1335. PubMed

Saborowski M, Kullak‐Ublick GA, Eloranta JJ (2006). The human organic cation transporter‐1 gene is transactivated by hepatocyte nuclear factor‐4alpha. J Pharmacol Exp Ther 317: 778–785. PubMed

Saini SP, Mu Y, Gong H, Toma D, Uppal H, Ren S et al. (2005). Dual role of orphan nuclear receptor pregnane X receptor in bilirubin detoxification in mice. Hepatology 41: 497–505. PubMed

Shu Y, Sheardown SA, Brown C, Owen RP, Zhang S, Castro RA et al. (2007). Effect of genetic variation in the organic cation transporter 1 (OCT1) on metformin action. J Clin Invest 117: 1422–1431. PubMed PMC

Smutny T, Bitman M, Urban M, Dubecka M, Vrzal R, Dvorak Z et al. (2014). U0126, a mitogen‐activated protein kinase kinase 1 and 2 (MEK1 and 2) inhibitor, selectively up‐regulates main isoforms of CYP3A subfamily via a pregnane X receptor (PXR) in HepG2 cells. Arch Toxicol 88: 2243–2259. PubMed

Wada T, Kang HS, Jetten AM, Xie W (2008). The emerging role of nuclear receptor RORalpha and its crosstalk with LXR in xeno‐ and endobiotic gene regulation. Exp Biol Med 233: 1191–1201. PubMed PMC

Wang DS, Jonker JW, Kato Y, Kusuhara H, Schinkel AH, Sugiyama Y (2002). Involvement of organic cation transporter 1 in hepatic and intestinal distribution of metformin. J Pharmacol Exp Ther 302: 510–515. PubMed

Wang DS, Kusuhara H, Kato Y, Jonker JW, Schinkel AH, Sugiyama Y (2003). Involvement of organic cation transporter 1 in the lactic acidosis caused by metformin. Mol Pharmacol 63: 844–848. PubMed

Wang JC, Stafford JM, Granner DK (1998). SRC‐1 and GRIP1 coactivate transcription with hepatocyte nuclear factor 4. J Biol Chem 273: 30847–30850. PubMed PMC

Watt AJ, Garrison WD, Duncan SA (2003). HNF4: a central regulator of hepatocyte differentiation and function. Hepatology 37: 1249–1253. PubMed

Zhai Y, Wada T, Zhang B, Khadem S, Ren S, Kuruba R et al. (2010). A functional cross‐talk between liver X receptor‐alpha and constitutive androstane receptor links lipogenesis and xenobiotic responses. Mol Pharmacol 78: 666–674. PubMed PMC

The hypolipidemic effect of MI-883, the combined CAR agonist/ PXR antagonist, in diet-induced hypercholesterolemia model

Discovery of Novel Human Constitutive Androstane Receptor Agonists with the Imidazo[1,2-a]pyridine Structure

Rifampicin Induces Gene, Protein, and Activity of P-Glycoprotein (ABCB1) in Human Precision-Cut Intestinal Slices

PLGA Based Nanospheres as a Potent Macrophage-Specific Drug Delivery System

The 3'-untranslated region contributes to the pregnane X receptor (PXR) expression down-regulation by PXR ligands and up-regulation by glucocorticoids

Teriflunomide Is an Indirect Human Constitutive Androstane Receptor (CAR) Activator Interacting With Epidermal Growth Factor (EGF) Signaling

Pregnane X Receptor (PXR)-Mediated Gene Repression and Cross-Talk of PXR with Other Nuclear Receptors via Coactivator Interactions