Isolation and characterization of DNA sequences amplified in multidrug-resistant hamster cells

P. Gros, James Croop, I. Roninson, A. Varshavsky, D. E. Housman

Research output: Contribution to journalArticle

155 Citations (Scopus)

Abstract

The mechanism by which mammalian cells acquire resistance to chemotherapeutic agents has been investigated by using molecular genetic techniques. LZ and C5, two independently derived multidrug-resistant Chinese hamster cell lines, share specific amplified DNA sequences. We demonstrate that commonly amplified DNA sequences reside in a contiguous domain of ≃120 kilobases (kb). We report the isolation of this DNA domain in cosmid clones and show that the level of amplification of the domain is correlated with the level of resistance in multidrug-resistant cell lines. The organization of the amplified domain was deduced by a unique approach utilizing in-gel hybridization of cloned DNA with amplified genomic DNA. We show that the entire cloned region is amplified in adriamycin-resistant LZ cells and independently derived, colchicine-resistant C5 cells. A mRNA species of ≃5 kb is encoded by a gene located within the boundaries of this region. Genomic sequences homologous to the 5-kb mRNA span over 75 kb of the amplified DNA segment. The level of expression of this mRNA in multidrug-resistant cells is correlated with the degree of gene amplification and the degree of drug resistance. Our results strongly suggest that the 5-kb mRNA species plays a role in the mechanism of multidrug resistance common to the LZ and C5 cell lines.

Original languageEnglish (US)
Pages (from-to)337-341
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume83
Issue number2
StatePublished - 1986
Externally publishedYes

Fingerprint

Cricetinae
Messenger RNA
DNA
Multiple Drug Resistance
Cell Line
Genetic Techniques
Cosmids
Gene Amplification
Colchicine
Sequence Homology
Cricetulus
Drug Resistance
Doxorubicin
Molecular Biology
Clone Cells
Gels
Genes

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

Isolation and characterization of DNA sequences amplified in multidrug-resistant hamster cells. / Gros, P.; Croop, James; Roninson, I.; Varshavsky, A.; Housman, D. E.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 83, No. 2, 1986, p. 337-341.

Research output: Contribution to journalArticle

@article{222c846827e24d3a89c4a150a2a886d9,
title = "Isolation and characterization of DNA sequences amplified in multidrug-resistant hamster cells",
abstract = "The mechanism by which mammalian cells acquire resistance to chemotherapeutic agents has been investigated by using molecular genetic techniques. LZ and C5, two independently derived multidrug-resistant Chinese hamster cell lines, share specific amplified DNA sequences. We demonstrate that commonly amplified DNA sequences reside in a contiguous domain of ≃120 kilobases (kb). We report the isolation of this DNA domain in cosmid clones and show that the level of amplification of the domain is correlated with the level of resistance in multidrug-resistant cell lines. The organization of the amplified domain was deduced by a unique approach utilizing in-gel hybridization of cloned DNA with amplified genomic DNA. We show that the entire cloned region is amplified in adriamycin-resistant LZ cells and independently derived, colchicine-resistant C5 cells. A mRNA species of ≃5 kb is encoded by a gene located within the boundaries of this region. Genomic sequences homologous to the 5-kb mRNA span over 75 kb of the amplified DNA segment. The level of expression of this mRNA in multidrug-resistant cells is correlated with the degree of gene amplification and the degree of drug resistance. Our results strongly suggest that the 5-kb mRNA species plays a role in the mechanism of multidrug resistance common to the LZ and C5 cell lines.",
author = "P. Gros and James Croop and I. Roninson and A. Varshavsky and Housman, {D. E.}",
year = "1986",
language = "English (US)",
volume = "83",
pages = "337--341",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
number = "2",

}

TY - JOUR

T1 - Isolation and characterization of DNA sequences amplified in multidrug-resistant hamster cells

AU - Gros, P.

AU - Croop, James

AU - Roninson, I.

AU - Varshavsky, A.

AU - Housman, D. E.

PY - 1986

Y1 - 1986

N2 - The mechanism by which mammalian cells acquire resistance to chemotherapeutic agents has been investigated by using molecular genetic techniques. LZ and C5, two independently derived multidrug-resistant Chinese hamster cell lines, share specific amplified DNA sequences. We demonstrate that commonly amplified DNA sequences reside in a contiguous domain of ≃120 kilobases (kb). We report the isolation of this DNA domain in cosmid clones and show that the level of amplification of the domain is correlated with the level of resistance in multidrug-resistant cell lines. The organization of the amplified domain was deduced by a unique approach utilizing in-gel hybridization of cloned DNA with amplified genomic DNA. We show that the entire cloned region is amplified in adriamycin-resistant LZ cells and independently derived, colchicine-resistant C5 cells. A mRNA species of ≃5 kb is encoded by a gene located within the boundaries of this region. Genomic sequences homologous to the 5-kb mRNA span over 75 kb of the amplified DNA segment. The level of expression of this mRNA in multidrug-resistant cells is correlated with the degree of gene amplification and the degree of drug resistance. Our results strongly suggest that the 5-kb mRNA species plays a role in the mechanism of multidrug resistance common to the LZ and C5 cell lines.

AB - The mechanism by which mammalian cells acquire resistance to chemotherapeutic agents has been investigated by using molecular genetic techniques. LZ and C5, two independently derived multidrug-resistant Chinese hamster cell lines, share specific amplified DNA sequences. We demonstrate that commonly amplified DNA sequences reside in a contiguous domain of ≃120 kilobases (kb). We report the isolation of this DNA domain in cosmid clones and show that the level of amplification of the domain is correlated with the level of resistance in multidrug-resistant cell lines. The organization of the amplified domain was deduced by a unique approach utilizing in-gel hybridization of cloned DNA with amplified genomic DNA. We show that the entire cloned region is amplified in adriamycin-resistant LZ cells and independently derived, colchicine-resistant C5 cells. A mRNA species of ≃5 kb is encoded by a gene located within the boundaries of this region. Genomic sequences homologous to the 5-kb mRNA span over 75 kb of the amplified DNA segment. The level of expression of this mRNA in multidrug-resistant cells is correlated with the degree of gene amplification and the degree of drug resistance. Our results strongly suggest that the 5-kb mRNA species plays a role in the mechanism of multidrug resistance common to the LZ and C5 cell lines.

UR - http://www.scopus.com/inward/record.url?scp=0022515908&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0022515908&partnerID=8YFLogxK

M3 - Article

C2 - 3455770

AN - SCOPUS:0022515908

VL - 83

SP - 337

EP - 341

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

SN - 0027-8424

IS - 2

ER -