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.
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 humanCAR 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.
Authors: T Kerbusch; J de Kraker; H J Keizer; J W van Putten; H J Groen; R L Jansen; J H Schellens; J H Beijnen Journal: Clin Pharmacokinet Date: 2001-01 Impact factor: 6.447
Authors: L B Moore; D J Parks; S A Jones; R K Bledsoe; T G Consler; J B Stimmel; B Goodwin; C Liddle; S G Blanchard; T M Willson; J L Collins; S A Kliewer Journal: J Biol Chem Date: 2000-05-19 Impact factor: 5.157
Authors: W Xie; J L Barwick; C M Simon; A M Pierce; S Safe; B Blumberg; P S Guzelian; R M Evans Journal: Genes Dev Date: 2000-12-01 Impact factor: 11.361
Authors: J Sugatani; H Kojima; A Ueda; S Kakizaki; K Yoshinari; Q H Gong; I S Owens; M Negishi; T Sueyoshi Journal: Hepatology Date: 2001-05 Impact factor: 17.425
Authors: Daniella Lowenberg; Caroline F Thorn; Zeruesenay Desta; David A Flockhart; Russ B Altman; Teri E Klein Journal: Pharmacogenet Genomics Date: 2014-02 Impact factor: 2.089
Authors: Duan Wang; Linhao Li; Hui Yang; Stephen S Ferguson; Maria R Baer; Ronald B Gartenhaus; Hongbing Wang Journal: Blood Date: 2012-11-16 Impact factor: 22.113
Authors: William D Hedrich; Jingwei Xiao; Scott Heyward; Yao Zhang; Junran Zhang; Maria R Baer; Hazem E Hassan; Hongbing Wang Journal: Mol Cancer Ther Date: 2016-01-28 Impact factor: 6.261