Effects of rifampicin on the expressions of drug transporter OATP1B1, drug metabolizing enzymes CYP3A4 and CYP3A5, and nuclear receptors PXR and AhR in human HepG2 and BeWo cells
Article Sidebar
Main Article Content
Abstract
Introduction: Rifampicin, an anti-tuberculosis drug, has been extensively reported for drug interaction due to its ability to induce several metabolizing enzymes. Drug transporter OATP1B1, drug metabolizing enzymes CYP3A4 and CYP3A5, and nuclear receptors PXR and AhR, are involved in metabolism of a clinical drug, especially in hepatocytes. However, the information of rifampicin on those genes in human placenta is still less. This study aimed to investigate the effects of rifampicin on the expressions of OATP1B1, CYP3A4 CYP3A5, PXR and AhR, in hepatocellular carcinoma (HepG2) and choriocarcinoma (BeWo) cells. Materials and method: HepG2 and BeWo cells (5x105 cells per well) were cultured in DMEM with 10% FBS and incubated with rifampicin at the final concentrations of 0.5, 5 and 10 mM for 24 hours. Total RNAs were extracted and the mRNA expression of each gene was determined by RT-qPCR. Results: Rifampicin induced the expression of CYP3A4 and CYP3A5 mRNA in both HepG2 and BeWo cells for 4-6 fold. Likewise, the levels of OATP1B1 mRNA were significantly elevated by rifampicin, compared to control group, in both HepG2 (18-23 fold) and BeWo (2-4 fold). However, rifampicin did not modify the expression of PXR in HepG2 while it extensively suppressed the PXR mRNA in BeWo. On the other hand, the expression of AhR was significantly induced by rifampicin in HepG2 but not in BeWo, only rifampicin at the lowest concentration test (0.5 mM) down-regulated the AhR mRNA expression (p-value < 0.05). Conclusion: The regulatory mechanism of rifampicin was different between HepG2 and BeWo cells. Rifampicin up-regulated the expression of OATP1B1, CYP3A4 and CYP3A5 in HepG2 cells, at least in part, via the AhR pathway. However, the expressions of OATP1B1, CYP3A4 and CYP3A5 in BeWo cells might be regulated by other receptors, beside PXR and AhR.
Article Details
In the case that some parts are used by others The author must Confirm that obtaining permission to use some of the original authors. And must attach evidence That the permission has been included
References
Abe T, Kakyo M, Tokui T, Nakagomi R, Nishio T, Nakai D, et al. Identification of a novel gene family encoding human liver-specific organic anion transporter LST-1. J Biol Chem 1999;274:17159–63.
Albrecht C, Soumian S, Tetlow N, Patel P, Sullivan MHF, Lakasing L, et al. Placental ABCA1 expression is reduced in primary antiphospholipid syndrome compared to pre-eclampsia and controls. Placenta 2007;28:701–8.
Benirschke K, Driscoll SG. The Pathology of the Human Placenta BT- Placenta. In: Strauss F, Benirschke K, Driscoll SG, editors., Berlin, Heidelberg: Springer Berlin Heidelberg; 1967, p. 97–571.
Bertilsson G, Heidrich J, Svensson K, Åsman M, Jendeberg L, Sydow-Bäckman M, et al. Identification of a human nuclear receptor defines a new signaling pathway for CYP3A4 induction. Proc Natl Acad Sci 1998;95:12208-13.
Blumberg B, Evans RM. Orphan nuclear receptors-new ligands and new possibilities. Genes Dev 1998;12:3149–55.
Blumberg B, Sabbagh W, Juguilon H, Bolado J, van Meter CM, Ong ES, et al. SXR, a novel steroid and xenobioticsensing nuclear receptor. Genes Dev 1998;12:3195–205.
Burk O, Koch I, Raucy J, Hustert E, Eichelbaum M, Brockmöller J, et al. The induction of cytochrome P450 3A5 (CYP3A5) in the human liver and intestine is mediated by the xenobiotic sensors pregnane X receptor (PXR) and constitutively activated receptor (CAR). J Biol Chem 2004;279:38379–85.
Chomczynski P, Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 1987;162:156–9.
Hagenbuch B, Gui C. Xenobiotic transporters of the human organic anion transporting polypeptides (OATP) family. Xenobiotica 2008;38:778–801.
Hakkola J, Pelkonen O, Pasanen M, Raunio H. Xenobiotic-metabolizing cytochrome P450 enzymes in the human feto-placental unit: role in intrauterine toxicity. Crit Rev Toxicol 1998;28:35–72.
Justenhoven C, Schaeffeler E, Winter S, Baisch C, Hamann U, Harth V, et al. Polymorphisms of the nuclear receptor pregnane X receptor and organic anion transporter polypeptides 1A2, 1B1, 1B3, and 2B1 are not associated with breast cancer risk. Breast Cancer Res Treat 2011;125:563–9.
König J, Cui Y, Nies AT, Keppler D. Localization and genomic organization of a new hepatocellular organic anion transporting polypeptide. J Biol Chem 2000;275:23161–8.
Lehmann JM, McKee DD, Watson MA, Willson TM, Moore JT, Kliewer SA. The human orphan nuclear receptor PXR is activated by compounds that regulate CYP3A4 gene expression and cause drug interactions. J Clin Invest 1998;102:1016–23.
Li T, Chiang JYL. Rifampicin induction of cyp3a4 requires pregnane x receptor cross talk with hepatocyte nuclear factor 4α and coactivators, and suppression of small heterodimer partner gene expression. Drug Metab Dispos 2006;34:756-64.
Luo G, Cunningham M, Kim S, Burn T, Lin J, Sinz M, et al. CYP3A4 Induction by drugs: Correlation between a Pregnane X Receptor reporter gene assay and CYP3A4 expression in human hepatocytes. Drug Metab Dispos 2002;30:795–804.
Lutz JD, Kirby BJ, Wang L, Song Q, Ling J, Massetto B, et al. Cytochrome P450 3A induction predicts p-glycoprotein induction; part 1: establishing induction relationships using ascending dose rifampin. Clin Pharmacol Ther 2018. doi:10.1002/cpt.1073.
McKinnon RA, Burgess WM, Hall PM, Roberts-Thomson SJ, Gonzalez FJ, McManus ME. Characterisation of CYP3A gene subfamily expression in human gastrointestinal tissues. Gut 1995;36:259-67.
Megan R, Amanda O, Bruno H. OATPs, OATs and OCTs: the organic anion and cation transporters of the SLCO and SLC22A gene superfamilies. Br J Pharmacol 2011;165:1260–87.
Meyer zu Schwabedissen HE, Kerstin B, Amarjit C, K. KH, G. SE, B. KR. Liver X receptor α and farnesoid X receptor are major transcriptional regulators of OATP1B1. Hepatology 2010;52:1797–807.
Moore JT, Kliewer SA. Use of the nuclear receptor PXR to predict drug interactions. Toxicology 2000;153:1–10.
Niemi M, Backman JT, Fromm MF, Neuvonen PJ, Kivistö KT. Pharmacokinetic interactions with rifampicin. Clin Pharmacokinet 2003;42:819–50.
Pichard-Garcia L, Hyland R, Baulieu J, Fabre JM, Milton A, Maurel P. Human hepatocytes in primary culture predict lack of cytochrome P-450 3A4 induction by eletriptan in vivo. Drug Metab Dispos 2000;28:51-7.
Poulsen MS, Rytting E, Mose T, Knudsen LE. Modeling placental transport: correlation of in vitro BeWo cell permeability and ex vivo human placental perfusion. Toxicol Vitr 2009;23:1380–6.
Rae JM, Johnson MD, Lippman ME, Flockhart DA. Rifampin is a selective, pleiotropic inducer of drug metabolism genes in human hepatocytes: studies with cdna and oligonucleotide expression arrays. J Pharmacol Exp Ther 2001;299:849-57.
Ramadoss P, Marcus C, Perdew GH. Role of the aryl hydrocarbon receptor in drug metabolism. Expert Opin Drug Metab Toxicol 2005;1:9–21.
Rasmussen MK, Daujat-Chavanieu M, Gerbal-Chaloin S. Activation of the aryl hydrocarbon receptor decreases rifampicin-induced CYP3A4 expression in primary human hepatocytes and HepaRG. Toxicol Lett 2017;277:1–8.
Schuetz EG, Schinkel AH, Relling M V, Schuetz JD. P-glycoprotein: a major determinant of rifampicin-inducible expression of cytochrome P4503A in mice and humans. Proc Natl Acad Sci 1996;93:4001-5.
Smythe W, Khandelwal A, Merle C, Rustomjee R, Gninafon M, Bocar Lo M, et al. A Semimechanistic pharmacokinetic-enzyme turnover model for rifampin autoinduction in adult tuberculosis patients. Antimicrob Agents Chemother 2012;56:2091–8.
Tirona RG, Leake BF, Wolkoff AW, Kim RB. Human organic anion transporting polypeptide-C (SLC21A6) is a major determinant of rifampin-mediated pregnane X receptor activation. J Pharmacol Exp Ther 2003;304:223-8.
Unadkat JD, Dahlin A, Vijay S. Placental Drug Transporters. Curr Drug Metab 2004;5:125–31.
Wang H, LeCluyse EL. Role of orphan nuclear receptors in the regulation of drug-metabolising enzymes. Clin Pharmacokinet 2003;42:1331–57.
Williamson B, Soars AC, Owen A, White P, Riley RJ, Soars MG. Dissecting the relative contribution of OATP1B1-mediated uptake of xenobiotics into human hepatocytes using siRNA. Xenobiotica 2013;43:920–31.
Xu C, Li CYT, Kong ANT. Induction of phase I, II and III drug metabolism/transport by xenobiotics. Arch Pharm Res 2005;28:249.
Yohei N, Ichiro I, Hiroshi S, Miyuki K, Kiyoshi K, Takeshi H, et al. Polymorphisms of OATP‐C (SLC21A6) and OAT3 (SLC22A8) genes: consequences for pravastatin pharmacokinetics. Clin Pharmacol Ther 2003;73:554–65.
Zhao L, Bin S, He HL, Yang JM, Pu YC, Gao CH, et al. Sodium butyrate increases P-gp expression in lung cancer by upregulation of STAT3 and mRNA stabilization of ABCB1. Anticancer Drugs 2018;29:227-33.