Evaluation and comparison of MRP1 activity with three fluorescent dyes and three modulators in leukemic cell lines.
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.
Dogan
A Lale
AL
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.
Legrand
Ollivier
O
Faussat
Anne-Marie
AM
Perrot
Jean-Yves
JY
Marie
Jean-Pierre
JP
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
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