Amplification of 4q21-q22 and the MXR gene in independently derived mitoxantrone-resistant cell lines

Turid Knutsen, V. Koneti Rao, Thomas Ried, Lyn Mickley, Erasmus Schneider, Keisuke Miyake, B. Michael Ghadimi, Hesed Padilla-Nash, Svetlana Pack, Lee Greenberger, Kenneth Cowan, Michael Dean, Tito Fojo, Susan Bates

Research output: Contribution to journalArticlepeer-review

78 Scopus citations

Abstract

Molecular cytogenetic studies were conducted on three multidrug- resistant cancer sublines which are highly resistant to the chemotherapeutic agent mitoxantrone, an anthracenedione. The three independently selected sublines were derived by exposure to mitoxantrone or Adriamycin and do not overexpress MDRI or MRP. Two sublines, MCF-7 AdVp3000 and MCF-7 MX, showed an amplification peak at 4q21-q22, as demonstrated by comparative genomic hybridization (CGH), while the third, SI-MI-80, did not. FISH using a whole chromosome 4 paint demonstrated multiple rearrangements involving chromosome 4 in MCF-7 AdVp3000 and MCF-7 MX, while SI-MI-80 contained only a simple reciprocal translocation. The parental cell lines had no chromosome 4 rearrangements and no copy number gain or amplification of chromosome 4. Spectral karyotyping (SKY) analysis revealed a balanced translocation, t(4;17)(q21-q22;p13) in SI-MI-80 and multiple clonal translocations involving chromosome 4 in MCF-7 AdVp3000 and MCF-7 MX. A novel cDNA, designated MXR, which encodes an ABC half-transporter and is highly overexpressed in the three sublines, was localized to chromosome 4 by somatic cell hybrid analysis. Southern blot analysis demonstrated amplification of the MXR gene in MCF-7 AdVp3000 and MCF-7 MX, but not in SI-MI-80. FISH studies with a BAC probe for MXR localized the gene to 4q21-22 in the normal chromosome 4 and revealed in both MCF-7 AdVp3000 and MCF-7 MX amplification of MXR at one translocation juncture, shown by SKY to be t(4;5)(4qter→4cen→4q21-22:: 5q13→5qter) in MCF-7 AdVp3000 and t(6;4;6;3)(6pter→6q15::4q21- q22::hsr::6q?::3q?27→3qter) in MCF MX; neither of the breakpoints in the partner chromosomes showed amplification by CGH. The data are consistent with the hypothesis of a transporter, presumably that encoded by the MXR gene, mediating mitoxantrone resistance. The MXR gene encodes a half-transporter and the absence of cytogenetic evidence of coamplification of other regions suggests that a partner may not be overexpressed, and instead the MXR half- transporter homodimerizes to mediate drug transport.

Original languageEnglish (US)
Pages (from-to)110-116
Number of pages7
JournalGenes Chromosomes and Cancer
Volume27
Issue number1
DOIs
StatePublished - Jan 2000
Externally publishedYes

ASJC Scopus subject areas

  • Genetics
  • Cancer Research

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