caa a22 4500 445313668 CHVBK 20180317142638.0 cr unu---uuuuu 170323e20110801xx s 000 0 eng 10.1007/s11095-011-0429-2 doi (NATIONALLICENCE)springer-10.1007/s11095-011-0429-2 The Role of Constitutive Androstane Receptor in Oxazaphosphorine-Mediated Induction of Drug-Metabolizing Enzymes in Human Hepatocytes [Elektronische Daten] [Duan Wang, Linhao Li, Jennifer Fuhrman, Stephen Ferguson, Hongbing Wang] ABSTRACT: Purpose: To investigate the roles of the constitutive androstane receptor (CAR) in cyclophosphamide (CPA)- and ifosfamide (IFO)-mediated induction of hepatic drug-metabolizing enzymes (DME). Methods: Induction of DMEs was evaluated using real-time RT-PCR and Western blotting analysis in human primary hepatocyte (HPH) cultures. Activation of CAR, pregnane X receptor (PXR), and aryl hydrocarbon receptor by CPA and IFO was assessed in cell-based reporter assays in HepG2 cells and/or nuclear translocation assays in HPHs. Results: CYP2B6 reporter activity was significantly enhanced by CPA and IFO in HepG2 cells co-transfected with CYP2B6 reporter plasmid and a chemical-responsive human CAR variant (CAR1 + A) construct. Real-time RT-PCR and Western blotting analysis in HPHs showed that both CPA and IFO induced the expressions of CYP2B6 and CYP3A4. Notably, treatment of HPHs with CPA but not IFO resulted in significant nuclear accumulation of CAR, which represents the initial step of CAR activation. Further studies in HPHs demonstrated that selective inhibition of PXR by sulforaphane preferentially repressed IFO- over CPA-mediated induction of CYP2B6. Conclusion: These results provide novel insights into the differential roles of CAR in the regulation of CPA- and IFO-induced DME expression and potential drug-drug interactions. Springer Science+Business Media, LLC, 2011 CAR nationallicence cyclophosphamide nationallicence CYP2B6 nationallicence ifosfamide nationallicence induction nationallicence 3MC : 3-methylcholanthrene nationallicence Ad/EYFP-hCAR : adenovirus expressing enhanced yellow fluorescent protein-tagged human CAR nationallicence AhR : aryl hydrocarbon receptor nationallicence CAR : constitutive androstane receptor nationallicence CITCO : 6-(4-chlorophenyl) imidazo[2,1-b][1,3]- thiazole-5-carbaldehyde-O-(3,4-dichlorobenzyl) oxime nationallicence CPA : cyclophosphamide nationallicence CYP : cytochrome P450 nationallicence DMSO : dimethyl sulfoxide nationallicence GAPDH : glyceraldehyde-3-phosphate dehydrogenase nationallicence HPH : human primary hepatocytes nationallicence IFO : ifosfamide nationallicence PB : phenobarbital nationallicence PK11195 : 1-(2-chlorophenyl-Nmethylpropyl)-3-isoquinoline-carboxamide nationallicence PXR : pregnane X receptor nationallicence RIF : rifampicin nationallicence RT-PCR : reverse transcription-polymerase chain reaction nationallicence SFN : sulforaphane nationallicence Wang Duan Department of Pharmaceutical Sciences, University of Maryland School of Pharmacy, 20 Penn Street, 21201, Baltimore, Maryland, USA aut Li Linhao Department of Pharmaceutical Sciences, University of Maryland School of Pharmacy, 20 Penn Street, 21201, Baltimore, Maryland, USA aut Fuhrman Jennifer Department of Pathology, University of Maryland School of Medicine, 21201, Baltimore, Maryland, USA aut Ferguson Stephen Life Technologies Corporation, 27707, Durham, North Carolina, USA aut Wang Hongbing Department of Pharmaceutical Sciences, University of Maryland School of Pharmacy, 20 Penn Street, 21201, Baltimore, Maryland, USA aut Pharmaceutical Research Springer US; http://www.springer-ny.com 28/8(2011-08-01), 2034-2044 0724-8741 28:8<2034 2011 28 11095 https://doi.org/10.1007/s11095-011-0429-2 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s11095-011-0429-2 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Wang Duan Department of Pharmaceutical Sciences, University of Maryland School of Pharmacy, 20 Penn Street, 21201, Baltimore, Maryland, USA aut NATIONALLICENCE 700 1- Li Linhao Department of Pharmaceutical Sciences, University of Maryland School of Pharmacy, 20 Penn Street, 21201, Baltimore, Maryland, USA aut NATIONALLICENCE 700 1- Fuhrman Jennifer Department of Pathology, University of Maryland School of Medicine, 21201, Baltimore, Maryland, USA aut NATIONALLICENCE 700 1- Ferguson Stephen Life Technologies Corporation, 27707, Durham, North Carolina, USA aut NATIONALLICENCE 700 1- Wang Hongbing Department of Pharmaceutical Sciences, University of Maryland School of Pharmacy, 20 Penn Street, 21201, Baltimore, Maryland, USA aut NATIONALLICENCE 773 0- Pharmaceutical Research Springer US; http://www.springer-ny.com 28/8(2011-08-01), 2034-2044 0724-8741 28:8<2034 2011 28 11095 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer