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Involvement of MDR 1 P‐glycoprotein in multifactorial resistance to methotrexate
Author(s) -
Norris Murray D.,
Graaf David De,
Haber Michelle,
Kavallaris Maria,
Madafiglio Janice,
Gilbert Jayne,
Kwan Edward,
Stewart Bernard W.,
Mechetner Eugene B.,
Gudkov Andrei V.,
Roninson Igor B.
Publication year - 1996
Publication title -
international journal of cancer
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.475
H-Index - 234
eISSN - 1097-0215
pISSN - 0020-7136
DOI - 10.1002/(sici)1097-0215(19960301)65:5<613::aid-ijc10>3.0.co;2-8
Subject(s) - methotrexate , glycoprotein , medicine , p glycoprotein , resistance (ecology) , immunology , cancer research , biology , oncology , genetics , drug resistance , multiple drug resistance , ecology
Cellular resistance to methotrexate (MTX) is believed to be unaffected by expression of MDRI P‐glycoprotein (Pgp), a pleiotropic efflux pump acting on different hydrophobic compounds that enter cells by passive diffusion. A series of human leukemic CCRF‐CEM sublines, isolated by multi‐step selection for very high resistance to MTX, exhibit multiple mechanisms of MTX resistance, including decreased carrier‐mediated uptake of MTX and DHFR gene amplification. These sublines show cross‐resistance to drugs of the multi‐drug resistance (MDR) family, which is correlated with relative resistance to MTX. The MTX‐selected sublines show increased expression and function of the MDRI gene, based on the measurement of MDRI mRNA, Pgp and rhodamine 123 accumulation. Sequence analysis of the MDRI cDNA from MTX‐selected CCRF‐CEM cells revealed no mutations in the protein coding region. MTX resistance in these cell lines is partially reversible by a Pgp‐specific monoclonal antibody (MAb) UIC2 and a monovalent Fab fragment of UIC2. Our results indicate that Pgp can contribute to multifactorial resistance to MTX. © 1996 Wiley‐Liss, Inc.