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Evaluation and comparison of MRP1 activity with three fluorescent dyes and three modulators in leukemic cell lines
A Lale Dogan1, Ollivier Legrand, Anne-Marie Faussat
1Laboratoire 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.
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.
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