PMID: 8600572Nov 1, 1995Paper

Acquisition of multiple copies of a mutant topoisomerase IIalpha allele by chromosome 17 aneuploidy is associated with etoposide resistance in human melanoma cell lines

Somatic Cell and Molecular Genetics
J A CampainI Pastan

Abstract

Mutants of the human melanoma cell line, FEM-X, selected in multiple steps with VP- 16 (etoposide), are cross resistant to the epipodophyllotoxins and doxorubicin. Complementary DNA's for topoisomerase IIalpha were cloned from both FEM-X and FVP3, the most resistant mutant. Deletion of nucleotides 1320-1322 (or Ala429 from the resulting topoisomerase IIalpha protein) was unique to the cDNA from the drug resistant cell line. Expression of the mutant mRNA increases in parallel with VP-16 resistance in this series of cell lines. Restriction analysis and Southern analysis with allele-specific oligonucleotide probes were used to quantify the ratio of wild-type to mutant topoisomerase IIalpha alleles present in DNA amplified by PCR from both FEM-X and the drug resistant sublines. This analysis shows that in cell lines of increasing drug resistance, the number of mutant topoisomerase IIalpha alleles increases incrementally along with a concomitant decrease in the number of wild-type alleles. By quantitative Southern analysis of genomic DNA the total number of topoisomerase IIalpha alleles in FVP3 is approximately 2-fold that in the parental cells. Fluorescence in situ hybridization with a chromosome l7 paint reveals that amplification...Continue Reading

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Citations

May 4, 2005·Biochemical and Biophysical Research Communications·Anette Duensing, Stefan Duensing
Mar 28, 1997·International Journal of Cancer. Journal International Du Cancer·W BergerM Micksche
Apr 27, 1999·Environmental and Molecular Mutagenesis·M S AlyS M El Nahas
Jul 31, 2001·International Journal of Cancer. Journal International Du Cancer·H HelmbachD Schadendorf
Jun 6, 2003·Oncogene·María S Soengas, Scott W Lowe
Aug 17, 2012·The Journal of Pharmacology and Experimental Therapeutics·Maarten E A ReithFranklin A Davis
Sep 4, 2015·PloS One·Mario BonatoMarco Zorzi
Nov 10, 2010·Journal of Speech, Language, and Hearing Research : JSLHR·Penny Anderson Gosselin, Jean-Pierre Gagné
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Aug 11, 2001·Drug Resistance Updates : Reviews and Commentaries in Antimicrobial and Anticancer Chemotherapy·William T. BeckUppoor G. Bhat

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