title: Establishment and use of new MDCK II cells overexpressing both UGT1A1 and MRP2 to characterize flavonoid metabolism via the glucuronidation pathway.
abstract: The purpose of this study is to characterize how overexpression of an efflux transporter and an UDP-glucuronosyltransferase (UGT) affects the cellular kinetics of glucuronidation processes.
  
  
  
  A new MDCK II cell line overexpressing both MRP2 and UGT1A1 (MDCKII-UGT1A1/MRP2 cells) was developed and used to determine how overexpression of an efflux transporter affects the kinetics of cellular flavonoid glucuronide production. The results showed that most model flavonoids (from a total of 13) were mainly metabolized into glucuronides in the MDCKII-UGT1A1/MRP2 cells and the glucuronides were rapidly excreted. Flavonoids with three or fewer hydroxyl group at 7, 3' or 6 hydroxyl group were also metabolized into sulfates. Mechanistic studies using 7-hydroxylflavone showed that its glucuronide was mainly (90%) effluxed by BCRP with a small (10%) but significant contribution from MRP2. Maximal velocity of glucuronide production MDCK-MRP2/UGT1A1 cells showed a fairly good correlation (R(2) >0.8) with those derived using UGT1A1 microsomes, but other kinetic parameters (e.g., Km ) did not correlate.
  
  
  
  Overexpression of a second efficient efflux transporter did not significantly change the fact that BCRP is the dominant transporter for flavonoid glucuronide nor did it diminish the influence of the efflux transporter as the "gate keeper" of glucuronidation process.
authors.affiliation: Hubei University of Medicine and University-Affiliated Taihe Hospital, Shiyan, China.|Department of Pharmacological and Pharmaceutical Sciences, College of Pharmacy, University of Houston, Houston, TX, USA.
authors.forename: Meifang
authors.initials: M
authors.lastname: Wang
authors.affiliation: Hubei University of Medicine and University-Affiliated Taihe Hospital, Shiyan, China.|Department of Pharmacological and Pharmaceutical Sciences, College of Pharmacy, University of Houston, Houston, TX, USA.|Hubei Provincial Technology and Research Center for Comprehensive Development of Medicinal Herbs, Hubei University of Medicine, Shiyan, Hubei, China.
authors.forename: Guangyi
authors.initials: G
authors.lastname: Yang
authors.affiliation: Department of Pharmacological and Pharmaceutical Sciences, College of Pharmacy, University of Houston, Houston, TX, USA.
authors.forename: Yu
authors.initials: Y
authors.lastname: He
authors.affiliation: Department of Pharmacological and Pharmaceutical Sciences, College of Pharmacy, University of Houston, Houston, TX, USA.
authors.forename: Beibei
authors.initials: B
authors.lastname: Xu
authors.affiliation: Hubei University of Medicine and University-Affiliated Taihe Hospital, Shiyan, China.|Department of Pharmacological and Pharmaceutical Sciences, College of Pharmacy, University of Houston, Houston, TX, USA.
authors.forename: Min
authors.initials: M
authors.lastname: Zeng
authors.affiliation: Department of Pharmacological and Pharmaceutical Sciences, College of Pharmacy, University of Houston, Houston, TX, USA.
authors.forename: Shufan
authors.initials: S
authors.lastname: Ge
authors.affiliation: Department of Pharmacological and Pharmaceutical Sciences, College of Pharmacy, University of Houston, Houston, TX, USA.
authors.forename: Taijun
authors.initials: T
authors.lastname: Yin
authors.affiliation: Hubei University of Medicine and University-Affiliated Taihe Hospital, Shiyan, China.|Department of Pharmacological and Pharmaceutical Sciences, College of Pharmacy, University of Houston, Houston, TX, USA.
authors.forename: Song
authors.initials: S
authors.lastname: Gao
authors.affiliation: Hubei University of Medicine and University-Affiliated Taihe Hospital, Shiyan, China. mhu@uh.edu.|Department of Pharmacological and Pharmaceutical Sciences, College of Pharmacy, University of Houston, Houston, TX, USA. mhu@uh.edu.
authors.forename: Ming
authors.initials: M
authors.lastname: Hu
mesh_terms: D000070997:ATP Binding Cassette Transporter, Subfamily G, Member 2 / Q000235:genetics / Q000378:metabolism; D000818:Animals; D004285:Dogs; D005419:Flavonoids / Q000378:metabolism / Q000493:pharmacokinetics*; D020719:Glucuronides / Q000378:metabolism*; D014453:Glucuronosyltransferase / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008658:Inactivation, Metabolic; D007700:Kinetics; D061985:Madin Darby Canine Kidney Cells; D000089762:Multidrug Resistance-Associated Protein 2; D027425:ATP-Binding Cassette, Sub-Family C Proteins / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D013431:Sulfates / Q000378:metabolism; D000099148:UGT1A1 Enzyme
chemical_list: D000070997:ATP Binding Cassette Transporter, Subfamily G, Member 2; D005419:Flavonoids; D020719:Glucuronides; D014453:Glucuronosyltransferase; D000089762:Multidrug Resistance-Associated Protein 2; D027425:ATP-Binding Cassette, Sub-Family C Proteins; D011994:Recombinant Proteins; D013431:Sulfates; D000099148:UGT1A1 Enzyme; C000715087:ABCC2 protein, human
keywords: Efflux transporters; Flavonoids; Glucuronide; UGT; “Revolving door”
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