title: Assessment of drug transporter function using fluorescent cell imaging.
abstract: ATP-binding cassette (ABC) proteins, including the breast cancer resistance protein (BCRP) and multidrug resistance proteins (MDRs), actively transport structurally diverse chemicals from a number of tissues and are being increasingly cited as mediators of clinically relevant drug-drug interactions. The potential outcomes of concomitantly administering two drugs that interact at the same transporter include altered disposition and toxicity and/or efficacy of one or both of the drugs. Research demonstrating the role of transporters in clinical pharmacokinetics has shed light on the need for in vitro screening methods that detect drug-transporter interactions during preclinical development. This unit describes cell-based procedures for detecting functional inhibitors of BCRP and MDR1 by measuring fluorescent substrate accumulation in suspended cells using an automated cell counter, which offers convenience, sensitivity, and speed in measuring intracellular fluorescence and identifying new inhibitors. An alternative method is provided for making similar measurements using a spectrophotometer with fluorescence detection capabilities.
authors: Department of Pharmacology and Toxicology, Ernest Mario School of Pharmacy, Rutgers University, Piscataway, New Jersey.
authors: Kristin M
authors: KM
authors: Bircsak
authors: Christopher J
authors: CJ
authors: Gibson
authors: Robert W
authors: RW
authors: Robey
authors: Lauren M
authors: LM
authors: Aleksunes
mesh_terms: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000037:antagonists & inhibitors*; D000070997:ATP Binding Cassette Transporter, Subfamily G, Member 2; D018528:ATP-Binding Cassette Transporters / Q000037:antagonists & inhibitors*; D001692:Biological Transport / Q000187:drug effects; D002460:Cell Line; D055808:Drug Discovery / Q000379:methods; D005456:Fluorescent Dyes / Q000737:chemistry*; D006801:Humans; D009363:Neoplasm Proteins / Q000037:antagonists & inhibitors*; D061848:Optical Imaging / Q000379:methods*
chemical_list: C116614:ABCG2 protein, human; D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D000070997:ATP Binding Cassette Transporter, Subfamily G, Member 2; D018528:ATP-Binding Cassette Transporters; D005456:Fluorescent Dyes; D009363:Neoplasm Proteins
keywords: ABC transporter; ABCB1; ABCG2; BCRP; MDR1