Active efflux of the free acid form of the fluorescent dye 2',7'-bis(2-carboxyethyl)-5(6)-carboxyfluorescein in multidrug-resistance-protein-overexpressing murine and human leukemia cells.
Murine and human cell lines overexpressing the multidrug-resistance protein (MRP) showed a marked decreased accumulation of the fluorescent dye 2',7'-bis(2-carboxyethyl)-5(6)-carboxyfluorescein (BCECF). In contrast, less altered accumulation was seen in the P-glycoprotein(P-gp)-overexpressing cell lines. The decreased drug accumulation was reversed by the energy inhibitors sodium azide/2-deoxyglucose and by the vinca alkaloid, vincristine, but not by the chemotherapeutic agents, etoposide and adriamycin. Decreased accumulation was linked to active efflux of the hydrophilic free acid form of BCECF from the MRP-overexpressing cell lines, indicating that dye extrusion occurs after the dye ester has been converted to the free acid form in the cytoplasm. The finding suggests that MRP mediates removal of substrates from a cytoplasmic location. Buthionine sulfoximine (BSO), an inhibitor of glutathione synthesis, decreased the vincristine and etoposide resistance displayed by the MRP-expressing murine cell lines, but did not affect the accumulation of BCECF. Thus, while glutathione may be involved in MRP-mediated resistance to some chemotherapeutic agents, it is not necessary for effiux of substrates such as BCECF.
Draper
M P
MP
Department of Molecular Biology, Tufts University School of Medicine, Boston, MA 02111, USA.
Martell
R L
RL
Levy
S B
SB
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000378:metabolism; D018528:ATP-Binding Cassette Transporters / Q000378:metabolism*; D000143:Acids; D000818:Animals; D001693:Biological Transport, Active / Q000187:drug effects; D019328:Buthionine Sulfoximine / Q000494:pharmacology; D002478:Cells, Cultured; D003593:Cytoplasm / Q000378:metabolism; D018432:Drug Resistance, Multiple; D005452:Fluoresceins / Q000737:chemistry / Q000378:metabolism*; D005978:Glutathione / Q000502:physiology; D006801:Humans; D007213:Indomethacin / Q000494:pharmacology; D051379:Mice; D027425:ATP-Binding Cassette, Sub-Family C Proteins; D014750:Vincristine / Q000494:pharmacology
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D018528:ATP-Binding Cassette Transporters; D000143:Acids; D019328:Buthionine Sulfoximine; D005452:Fluoresceins; D005978:Glutathione; D007213:Indomethacin; D027425:ATP-Binding Cassette, Sub-Family C Proteins; D014750:Vincristine; C043829:2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein