ATP- and glutathione-dependent transport of chemotherapeutic drugs by the multidrug resistance protein MRP1.
The present study was performed to investigate the ability of the multidrug resistance protein (MRPI) to transport different cationic substrates in comparison with MDR1-P-glycoprotein (MDR1). Transport studies were performed with isolated membrane vesicles from in vitro selected multidrug resistant cell lines overexpressing MDR1 (A2780AD) or MRP1 (GLC4/Adr) and a MRP1-transfected cell line (S1(MRP)). As substrates we used 3H-labelled derivatives of the hydrophilic monoquaternary cation N-(4',4'-azo-in-pentyl)-21-deoxy-ajmalinium (APDA), the basic drug vincristine and the more hydrophobic basic drug daunorubicin. All three are known MDR1-substrates. MRP1 did not mediate transport of these substrates per se. In the presence of reduced glutathione (GSH), there was an ATP-dependent uptake of vincristine and daunorubicin, but not of APDA, into GLC4/Adr and S1(MRP) membrane vesicles which could be inhibited by the MRP1-inhibitor MK571. ATP- and GSH-dependent transport of daunorubicin and vincristine into GLC4/Adr membrane vesicles was inhibited by the MRP1-specific monoclonal antibody QCRL-3. MRP1-mediated daunorubicin transport rates were dependent on the concentration of GSH and were maximal at concentrations > or = 10 mM. The apparent KM value for GSH was 2.7 mM. Transport of daunorubicin in the presence of 10 mM GSH was inhibited by MK571 with an IC50 of 0.4 microM. In conclusion, these results demonstrate that MRP1 transports vincristine and daunorubicin in an ATP- and GSH-dependent manner. APDA is not a substrate for MRP1.
Renes
J
J
Groningen Institute for Drug Studies, Division of Gastroenterology and Hepatology, University Hospital Groningen, The Netherlands. j.w.renes@med.rug.nl
de Vries
E G
EG
Nienhuis
E F
EF
Jansen
P L
PL
Müller
M
M
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000378:metabolism; D018528:ATP-Binding Cassette Transporters / Q000276:immunology / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000494:pharmacology*; D000404:Ajmaline / Q000031:analogs & derivatives / Q000493:pharmacokinetics; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000494:pharmacology; D000970:Antineoplastic Agents / Q000494:pharmacology*; D001692:Biological Transport / Q000187:drug effects; D002412:Cations / Q000493:pharmacokinetics; D003630:Daunorubicin / Q000493:pharmacokinetics; D004305:Dose-Response Relationship, Drug; D005978:Glutathione / Q000494:pharmacology*; D006801:Humans; D015151:Immunoblotting; D017997:Leukotriene C4 / Q000493:pharmacokinetics; D008566:Membranes / Q000737:chemistry / Q000378:metabolism; D027425:ATP-Binding Cassette, Sub-Family C Proteins; D014316:Tritium; D014407:Tumor Cells, Cultured; D014750:Vincristine / Q000493:pharmacokinetics
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D018528:ATP-Binding Cassette Transporters; D000255:Adenosine Triphosphate; D000404:Ajmaline; D000911:Antibodies, Monoclonal; D000970:Antineoplastic Agents; D002412:Cations; D003630:Daunorubicin; D005978:Glutathione; D017997:Leukotriene C4; D027425:ATP-Binding Cassette, Sub-Family C Proteins; D014316:Tritium; D014750:Vincristine