Abstract
U-937 human leukemia cells were selected for resistance to doxorubicin in the presence or absence of a specific drug modulator that inhibits the activity of P-glycoprotein (Pgp), encoded by the multidrug-resistance gene (MDR1). Parental cells expressed low basal levels of the multidrug-resistance-associated gene (MRP1) and major vault protein (MVP) mRNAs and no MDR1 mRNA. Two doxorubicin-resistant cell lines were selected. Both drug-resistant cell lines upregulated the MVP mRNA level 1.5-fold within 1 cell passage. The MVP mRNA level continued to increase over time as the doxorubicin selection pressure was increased. MVP protein levels generally paralleled the mRNA levels. The 2 high molecular weight vault protein mRNAs were always expressed at constitutive levels. Fully formed vault particles consisting of the MVP, the 2 high molecular weight proteins and the vault RNA assembled and accumulated to increased levels in drug-selected cells. MVP induction is therefore the rate-limiting step for vault particle formation in U-937 cells. By passage 25 and thereafter, the selected cells were resistant to doxorubicin, etoposide, mitoxantrone and 5-fluorouracil by a pathway that was independent of MDR1, MRP1, MRP2 and breast cancer resistance protein. In summary, U-937 doxorubicin-selected cells are programmed to rapidly upregulate MVP mRNA levels, to accumulate vault particles and to become multidrug resistant.
Copyright 2002 Wiley-Liss, Inc.
MeSH terms
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ATP Binding Cassette Transporter, Subfamily B, Member 1 / genetics
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ATP Binding Cassette Transporter, Subfamily B, Member 1 / metabolism
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ATP Binding Cassette Transporter, Subfamily G, Member 2
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ATP-Binding Cassette Transporters / genetics
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ATP-Binding Cassette Transporters / metabolism
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Antibiotics, Antineoplastic / pharmacology*
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Antineoplastic Agents / pharmacology
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Cell Compartmentation
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Doxorubicin / pharmacology*
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Drug Resistance, Neoplasm*
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Heat-Shock Response
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Hot Temperature
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Humans
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Mitochondrial Proteins*
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Multidrug Resistance-Associated Proteins / genetics
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Multidrug Resistance-Associated Proteins / metabolism
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Neoplasm Proteins*
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Precipitin Tests
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RNA, Messenger / metabolism
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RNA, Neoplasm / metabolism
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Reverse Transcriptase Polymerase Chain Reaction
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Ribosomal Proteins / genetics
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Ribosomal Proteins / metabolism
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Saccharomyces cerevisiae Proteins*
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U937 Cells / drug effects*
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U937 Cells / metabolism
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Up-Regulation
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Vault Ribonucleoprotein Particles / biosynthesis*
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Vault Ribonucleoprotein Particles / genetics
Substances
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ABCG2 protein, human
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ATP Binding Cassette Transporter, Subfamily B, Member 1
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ATP Binding Cassette Transporter, Subfamily G, Member 2
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ATP-Binding Cassette Transporters
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Antibiotics, Antineoplastic
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Antineoplastic Agents
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MRP2 protein, S cerevisiae
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Mitochondrial Proteins
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Multidrug Resistance-Associated Proteins
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Neoplasm Proteins
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RNA, Messenger
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RNA, Neoplasm
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Ribosomal Proteins
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Saccharomyces cerevisiae Proteins
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Vault Ribonucleoprotein Particles
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major vault protein
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Doxorubicin
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multidrug resistance-associated protein 1