Activity and expression of the multidrug resistance proteins MRP1 and MRP2 in acute myeloid leukemia cells, tumor cell lines, and normal hematopoietic CD34+ peripheral blood cells.
To develop a functional assay for the activity of the multidrug resistance protein 1 (MRP1), we tested whether carboxyfluorescein (CF) was specifically transported by MRP and whether this transport pump could be specifically blocked by the leukotriene D4 receptor antagonist MK-571. The activity and expression of MRP1 were studied in several tumor cell lines and in leukemic blasts from patients with acute myeloid leukemia (AML). In the MRP1-overexpressing cell line GLC4/ADR and the MRP1-transfected cell line S1(MRP), MK-571 inhibited CF efflux with high factors [45.9+/-5.8 (mean+/-SD) and 14.4+/-3.2, respectively; n=3; efflux-blocking factors are defined as the ratio of the median fluorescence in presence or absence of MK-571] compared with their MRP1 low-expressing counterparts GLC4 (11.5+/-2.7) and S1 (2.8+/-0.4). In 15 AML cases, the CF efflux-blocking factors of MK-571 varied between 1.9 and 5.2. A good correlation was found between MRP1 protein expression and CF efflux-blocking factors of MK-571 (P=0.013, r=0.39). Besides MRP1, MRP2 was demonstrated with reverse transcription-PCR in 40% of the cases. In contrast to the cell lines, MK-571 also inhibited rhodamine 123 (Rh123) efflux in AML samples. On the other hand, PSC833, a P-glycoprotein specific inhibitor, did not modulate the CF efflux but is efficient in blocking Rh123 efflux. This study demonstrates that AML blasts express MRP1 and MRP2 and that MRP1 activity can be determined by a flow cytometric assay using CF and MK-571.
van der Kolk
D M
DM
Department of Internal Medicine, University Hospital of Groningen, The Netherlands.
de Vries
E G
EG
Koning
J A
JA
van den Berg
E
E
Müller
M
M
Vellenga
E
E
D018528:ATP-Binding Cassette Transporters / Q000378:metabolism*; D000208:Acute Disease; D019008:Drug Resistance, Neoplasm; D005434:Flow Cytometry; D006801:Humans; D007951:Leukemia, Myeloid / Q000378:metabolism*; D027425:ATP-Binding Cassette, Sub-Family C Proteins; D009363:Neoplasm Proteins / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000378:metabolism; D014407:Tumor Cells, Cultured / Q000187:drug effects
D018528:ATP-Binding Cassette Transporters; D027425:ATP-Binding Cassette, Sub-Family C Proteins; D009363:Neoplasm Proteins; D012333:RNA, Messenger