Comparative M-FISH and CGH analyses in sensitive and drug-resistant human T-cell acute leukemia cell lines

Abstract

Cell lines of human T-cell acute lymphoblastic leukemias (T-ALL) have gained high interest for study of mechanisms of cytostatic drug resistance. However, they should also be suited to examine the validity and reliability of molecular cytogenetic techniques in detecting genomic alterations in neoplastic cells. Therefore, comparative genomic hybridization (CGH) and 24-color-fluorescence-in-situ-hybridization (M-FISH) were applied to eight sublines of CCRF-CEM leukemia cells selected in vitro for drug resistance and to their drug-sensitive parental counterparts. All cell lines were characterized by altered chromosome numbers and by a variety of chromosomal structural aberrations as shown by M-FISH. The great majority of anomalies detected by this technique were confirmed by CGH. Interestingly, a considerable number of the rearrangements found were imbalanced. Amplifications of 5q13 in the six methotrexate-resistant cell lines, a del(9)(p21pter) in all lines examined, and a gain of chromosome 20 in 9 of the 10 lines examined were readily detected by both techniques. The same held true for losses of chromosomes 17 and 18 in the near tetraploid cell lines which could also be confirmed by CGH. Some imbalances of genomic material detected by CGH were, however, not observed by means of M-FISH, possibly due to the limited extension of the corresponding chromosomal segment involved or the small subpopulation of cells affected. On the other hand, reciprocal translocations, balanced isochromosomes, and small deletions remained mainly undetected by CGH. A comparison of chromosomal alterations in drug-resistant and parental cell lines showed not only amplifications of chromosomal segments harboring well-known drug resistance genes, e.g., the dihydrofolate reductase gene, but also chromosomal changes which may involve novel genes associated with drug resistance. Thus, the present study has clearly unveiled the strengths and weaknesses of both techniques which can excellently complement each other. Their combination allowed a distinct improvement of the definition of the complex karyotypes of drug-resistant cell lines.

Autoren

A Weise
T Liehr
T Efferth
A Kuechler
E Gebhart

DOI

10.1159/000069808

eISSN

1424-859X

ISSN

1424-8581

Ausgabe der Veröffentlichung

2-3

Zeitschrift

Cytogenetic and Genome Research

Sprache

en

Online publication date

2003

Paginierung

118 - 125

Datum der Veröffentlichung

2002

Status

Published

Herausgeber

S. Karger AG

Herausgeber URL

http://dx.doi.org/10.1159/000069808

Datum der Datenerfassung

2022

Titel

Comparative M-FISH and CGH analyses in sensitive and drug-resistant human T-cell acute leukemia cell lines

Ausgabe der Zeitschrift

98

Data source: Crossref

Abstract

Cell lines of human T-cell acute lymphoblastic leukemias (T-ALL) have gained high interest for study of mechanisms of cytostatic drug resistance. However, they should also be suited to examine the validity and reliability of molecular cytogenetic techniques in detecting genomic alterations in neoplastic cells. Therefore, comparative genomic hybridization (CGH) and 24-color-fluorescence-in-situ-hybridization (M-FISH) were applied to eight sublines of CCRF-CEM leukemia cells selected in vitro for drug resistance and to their drug-sensitive parental counterparts. All cell lines were characterized by altered chromosome numbers and by a variety of chromosomal structural aberrations as shown by M-FISH. The great majority of anomalies detected by this technique were confirmed by CGH. Interestingly, a considerable number of the rearrangements found were imbalanced. Amplifications of 5q13 in the six methotrexate-resistant cell lines, a del(9)(p21pter) in all lines examined, and a gain of chromosome 20 in 9 of the 10 lines examined were readily detected by both techniques. The same held true for losses of chromosomes 17 and 18 in the near tetraploid cell lines which could also be confirmed by CGH. Some imbalances of genomic material detected by CGH were, however, not observed by means of M-FISH, possibly due to the limited extension of the corresponding chromosomal segment involved or the small subpopulation of cells affected. On the other hand, reciprocal translocations, balanced isochromosomes, and small deletions remained mainly undetected by CGH. A comparison of chromosomal alterations in drug-resistant and parental cell lines showed not only amplifications of chromosomal segments harboring well-known drug resistance genes, e.g., the dihydrofolate reductase gene, but also chromosomal changes which may involve novel genes associated with drug resistance. Thus, the present study has clearly unveiled the strengths and weaknesses of both techniques which can excellently complement each other. Their combination allowed a distinct improvement of the definition of the complex karyotypes of drug-resistant cell lines.

Addresses

Institute of Human Genetics and Anthropology, Friedrich Schiller University, Jena, Germany.

Autoren

A Weise
T Liehr
T Efferth
T Efferth
A Kuechler
E Gebhart

DOI

10.1159/000069808

eISSN

1424-859X

Externe Identifier

PubMed Identifier: 12697993

Open access

false

ISSN

1424-8581

Ausgabe der Veröffentlichung

2-3

Zeitschrift

Cytogenetic and genome research

Schlüsselwörter

Tumor Cells, Cultured
Chromosomes, Human
Humans
Chromosome Aberrations
In Situ Hybridization, Fluorescence
Nucleic Acid Hybridization
Drug Resistance, Neoplasm
Leukemia-Lymphoma, Adult T-Cell

Sprache

eng

Medium

Print

Paginierung

118 - 125

Datum der Veröffentlichung

2002

Status

Published

Datum der Datenerfassung

2003

Titel

Comparative M-FISH and CGH analyses in sensitive and drug-resistant human T-cell acute leukemia cell lines.

Sub types

Comparative Study
Research Support, Non-U.S. Gov't
Evaluation Study
Journal Article

Ausgabe der Zeitschrift

98

Data source: Europe PubMed Central

Abstract

Cell lines of human T-cell acute lymphoblastic leukemias (T-ALL) have gained high interest for study of mechanisms of cytostatic drug resistance. However, they should also be suited to examine the validity and reliability of molecular cytogenetic techniques in detecting genomic alterations in neoplastic cells. Therefore, comparative genomic hybridization (CGH) and 24-color-fluorescence-in-situ-hybridization (M-FISH) were applied to eight sublines of CCRF-CEM leukemia cells selected in vitro for drug resistance and to their drug-sensitive parental counterparts. All cell lines were characterized by altered chromosome numbers and by a variety of chromosomal structural aberrations as shown by M-FISH. The great majority of anomalies detected by this technique were confirmed by CGH. Interestingly, a considerable number of the rearrangements found were imbalanced. Amplifications of 5q13 in the six methotrexate-resistant cell lines, a del(9)(p21pter) in all lines examined, and a gain of chromosome 20 in 9 of the 10 lines examined were readily detected by both techniques. The same held true for losses of chromosomes 17 and 18 in the near tetraploid cell lines which could also be confirmed by CGH. Some imbalances of genomic material detected by CGH were, however, not observed by means of M-FISH, possibly due to the limited extension of the corresponding chromosomal segment involved or the small subpopulation of cells affected. On the other hand, reciprocal translocations, balanced isochromosomes, and small deletions remained mainly undetected by CGH. A comparison of chromosomal alterations in drug-resistant and parental cell lines showed not only amplifications of chromosomal segments harboring well-known drug resistance genes, e.g., the dihydrofolate reductase gene, but also chromosomal changes which may involve novel genes associated with drug resistance. Thus, the present study has clearly unveiled the strengths and weaknesses of both techniques which can excellently complement each other. Their combination allowed a distinct improvement of the definition of the complex karyotypes of drug-resistant cell lines.

Date of acceptance

2002

Autoren

A Weise
T Liehr
T Efferth
A Kuechler
E Gebhart

Autoren-URL

https://www.ncbi.nlm.nih.gov/pubmed/12697993

DOI

10.1159/000069808

ISSN

1424-8581

Ausgabe der Veröffentlichung

2-3

Zeitschrift

Cytogenet Genome Res

Schlüsselwörter

Chromosome Aberrations
Chromosomes, Human
Drug Resistance, Neoplasm
Humans
In Situ Hybridization, Fluorescence
Leukemia-Lymphoma, Adult T-Cell
Nucleic Acid Hybridization
Tumor Cells, Cultured

Sprache

eng

Country

Switzerland

Paginierung

118 - 125

PII

69808

Datum der Veröffentlichung

2002

Status

Published

Datum, an dem der Datensatz öffentlich gemacht wurde

2003

Titel

Comparative M-FISH and CGH analyses in sensitive and drug-resistant human T-cell acute leukemia cell lines.

Sub types

Comparative Study
Evaluation Study
Journal Article
Research Support, Non-U.S. Gov't

Ausgabe der Zeitschrift

98

Data source: PubMed

Comparative M-FISH and CGH analyses in sensitive and drug-resistant human T-cell acute leukemia cell lines

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