title: Efflux kinetics and intracellular distribution of daunorubicin are not affected by major vault protein/lung resistance-related protein (vault) expression.
abstract: Vaults may contribute to multidrug resistance by transporting drugs away from their subcellular targets. To study the involvement of vaults in the extrusion of anthracyclines from the nucleus, we investigated the handling of daunorubicin by drug-sensitive and drug-resistant non-small lung cancer cells, including a green fluorescent protein (GFP)-tagged major vault protein (MVP)-overexpressing transfectant (SW1573/MVP-GFP). Cells were exposed to 1 microm daunorubicin for 60 min, after which the cells were allowed to efflux the accumulated drug. No significant differences in daunorubicin efflux kinetics were observed between the sensitive SW1573 and SW1573/MVP-GFP transfectant, whereas the drug-resistant SW1573/2R120 cells clearly demonstrated an increased efflux rate. It was noted that the redistribution of daunorubicin from the nucleus into distinct vesicular structures in the cytoplasm was not accompanied by changes in the intracellular localization of vaults. Similar experiments were performed using mouse embryonic fibroblasts derived from wild-type and MVP knockout mice, which were previously shown to be devoid of vault particles. Both cell lines showed comparable drug efflux rates, and the intracellular distribution of daunorubicin in time was identical. Reintroduction of a human MVP tagged with GFP in the MVP(-/-) cells results in the formation of vault particles but did not give rise an altered daunorubicin handling compared with MVP(-/-) cells expressing GFP. Our results indicate that vaults are not directly involved in the sequestration of anthracyclines in vesicles nor in their efflux from the nucleus.
authors.affiliation: Department of Hematology, Erasmus Medical Center, 3000 DR Rotterdam, The Netherlands.
authors.forename: Arend
authors.initials: A
authors.lastname: van Zon
authors.forename: Marieke H
authors.initials: MH
authors.lastname: Mossink
authors.forename: Martijn
authors.initials: M
authors.lastname: Schoester
authors.forename: Rik J
authors.initials: RJ
authors.lastname: Scheper
authors.forename: Pieter
authors.initials: P
authors.lastname: Sonneveld
authors.forename: Erik A C
authors.initials: EA
authors.lastname: Wiemer
mesh_terms: D000818:Animals; D000903:Antibiotics, Antineoplastic / Q000493:pharmacokinetics*; D002289:Carcinoma, Non-Small-Cell Lung / Q000188:drug therapy / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D003630:Daunorubicin / Q000493:pharmacokinetics*; D018432:Drug Resistance, Multiple; D019008:Drug Resistance, Neoplasm; D005260:Female; D005347:Fibroblasts / Q000378:metabolism; D049452:Green Fluorescent Proteins; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics / Q000493:pharmacokinetics; D008175:Lung Neoplasms / Q000188:drug therapy / Q000235:genetics / Q000378:metabolism*; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000493:pharmacokinetics; D014162:Transfection; D020394:Vault Ribonucleoprotein Particles / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D000099187:Major Vault Protein
chemical_list: D000903:Antibiotics, Antineoplastic; D003630:Daunorubicin; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D020394:Vault Ribonucleoprotein Particles; D000099187:Major Vault Protein