title: Evaluation and comparison of MRP1 activity with three fluorescent dyes and three modulators in leukemic cell lines.
abstract: MRP1 activity was evaluated and compared in 11 cell lines with different levels of MRP1 expression using functional assays of calcein acetoxymethyl ester (calcein-AM), carboxyfluorescein diacetate (CFDA) and Rhodamine 123 (Rh123) in combination with the modulators cyclosporin A (CsA), probenecid and MK571. A good correlation was found between MRP1 expression and the modulatory effect of MK571 on calcein-AM uptake (P = 0.01 and probenecid effect on CFDA uptake (P = 0.02). Additionally, the combined modulatory effect of MK571 and probenecid on CFDA uptake (P < 0.0001) and on calcein-AM uptake (P = 0.0001) were highly significant. No correlation was found between MRP1 expression and the effects of three modulators on Rh123 uptake or efflux. In conclusion, calcein-AM and CFDA uptake assays are the best choices to probe MRP1 activity and combination of two modulators may improve the efficiency of these assays.
authors.affiliation: Laboratoire INSERM E03-55/EA 1529, Département d'Onco-Hématologie, Hôpital Hôtel-Dieu AP-HP, Universite Paris 6, 1 Place du Parvis Notre Dame, Paris 75004, France.
authors.forename: A Lale
authors.initials: AL
authors.lastname: Dogan
authors.forename: Ollivier
authors.initials: O
authors.lastname: Legrand
authors.forename: Anne-Marie
authors.initials: AM
authors.lastname: Faussat
authors.forename: Jean-Yves
authors.initials: JY
authors.lastname: Perrot
authors.forename: Jean-Pierre
authors.initials: JP
authors.lastname: Marie
mesh_terms: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000378:metabolism; D001692:Biological Transport / Q000187:drug effects; D001993:Bronchodilator Agents / Q000494:pharmacology*; D016572:Cyclosporine / Q000494:pharmacology; D018432:Drug Resistance, Multiple; D019008:Drug Resistance, Neoplasm; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005452:Fluoresceins / Q000493:pharmacokinetics; D005456:Fluorescent Dyes / Q000493:pharmacokinetics*; D006801:Humans; D007938:Leukemia / Q000378:metabolism* / Q000473:pathology*; D027425:ATP-Binding Cassette, Sub-Family C Proteins / Q000378:metabolism*; D011339:Probenecid / Q000494:pharmacology; D011422:Propionates / Q000494:pharmacology; D011804:Quinolines / Q000494:pharmacology; D020112:Rhodamine 123 / Q000493:pharmacokinetics; D014407:Tumor Cells, Cultured
chemical_list: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D001993:Bronchodilator Agents; D016572:Cyclosporine; D004791:Enzyme Inhibitors; D005452:Fluoresceins; D005456:Fluorescent Dyes; D027425:ATP-Binding Cassette, Sub-Family C Proteins; D011339:Probenecid; D011422:Propionates; D011804:Quinolines; D020112:Rhodamine 123; C085925:calcein AM; C024098:6-carboxyfluorescein; C059141:verlukast; C421109:multidrug resistance-associated protein 1