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Incidence of Mutation and Deletion in Topoisomerase IIα mRNA of Etoposide and mAMSA–resistant Cell Lines

The efficacy of all chemotherapeutic agents is limited by the occurrence of drug resistance. To further understand resistance to topoisomerase II inhibitors, 50 sublines were isolated as single clones from parental cells by exposure to VP–16 (etoposide) or mAMSA (m–amsacrine). Subsequently, a popula...

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Autores principales: Matsumoto, Yoshihito, Takano, Hiroshi, Kunishio, Katsuzo, Nagao, Seigo, Fojo, Tito
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Blackwell Publishing Ltd 2001
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5926608/
https://www.ncbi.nlm.nih.gov/pubmed/11676865
http://dx.doi.org/10.1111/j.1349-7006.2001.tb01069.x
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author Matsumoto, Yoshihito
Takano, Hiroshi
Kunishio, Katsuzo
Nagao, Seigo
Fojo, Tito
author_facet Matsumoto, Yoshihito
Takano, Hiroshi
Kunishio, Katsuzo
Nagao, Seigo
Fojo, Tito
author_sort Matsumoto, Yoshihito
collection PubMed
description The efficacy of all chemotherapeutic agents is limited by the occurrence of drug resistance. To further understand resistance to topoisomerase II inhibitors, 50 sublines were isolated as single clones from parental cells by exposure to VP–16 (etoposide) or mAMSA (m–amsacrine). Subsequently, a population of cells from each subline was exposed to three–fold higher drug concentrations allowing 16 stable sublines to be established at higher extracellular drug concentration. Finally, 66 sublines were picked up. The frequency and nature of mutations in the topoisomerase II gene in the drug–selected cell lines were evaluated. In order to screen a large number of cell lines, an RNAse protection assay was developed and mismatches were observed in 13.6% of resistant cell lines (12% of resistant cell lines exposed to lower drug concentrations and 18.8% of resistant cell lines exposed to higher drug concentrations). Some of these mutations are located in vital regions of topoisomerase II (phosphorylation sites in the C–terminal or N–terminal, and nuclear localizing signal of topoisomerase II). Our findings suggest that mutations of topoisomerase II gene are an important and frequent mechanism of resistance to topoisomerase II inhibitors.
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spelling pubmed-59266082018-05-11 Incidence of Mutation and Deletion in Topoisomerase IIα mRNA of Etoposide and mAMSA–resistant Cell Lines Matsumoto, Yoshihito Takano, Hiroshi Kunishio, Katsuzo Nagao, Seigo Fojo, Tito Jpn J Cancer Res Article The efficacy of all chemotherapeutic agents is limited by the occurrence of drug resistance. To further understand resistance to topoisomerase II inhibitors, 50 sublines were isolated as single clones from parental cells by exposure to VP–16 (etoposide) or mAMSA (m–amsacrine). Subsequently, a population of cells from each subline was exposed to three–fold higher drug concentrations allowing 16 stable sublines to be established at higher extracellular drug concentration. Finally, 66 sublines were picked up. The frequency and nature of mutations in the topoisomerase II gene in the drug–selected cell lines were evaluated. In order to screen a large number of cell lines, an RNAse protection assay was developed and mismatches were observed in 13.6% of resistant cell lines (12% of resistant cell lines exposed to lower drug concentrations and 18.8% of resistant cell lines exposed to higher drug concentrations). Some of these mutations are located in vital regions of topoisomerase II (phosphorylation sites in the C–terminal or N–terminal, and nuclear localizing signal of topoisomerase II). Our findings suggest that mutations of topoisomerase II gene are an important and frequent mechanism of resistance to topoisomerase II inhibitors. Blackwell Publishing Ltd 2001-10 /pmc/articles/PMC5926608/ /pubmed/11676865 http://dx.doi.org/10.1111/j.1349-7006.2001.tb01069.x Text en
spellingShingle Article
Matsumoto, Yoshihito
Takano, Hiroshi
Kunishio, Katsuzo
Nagao, Seigo
Fojo, Tito
Incidence of Mutation and Deletion in Topoisomerase IIα mRNA of Etoposide and mAMSA–resistant Cell Lines
title Incidence of Mutation and Deletion in Topoisomerase IIα mRNA of Etoposide and mAMSA–resistant Cell Lines
title_full Incidence of Mutation and Deletion in Topoisomerase IIα mRNA of Etoposide and mAMSA–resistant Cell Lines
title_fullStr Incidence of Mutation and Deletion in Topoisomerase IIα mRNA of Etoposide and mAMSA–resistant Cell Lines
title_full_unstemmed Incidence of Mutation and Deletion in Topoisomerase IIα mRNA of Etoposide and mAMSA–resistant Cell Lines
title_short Incidence of Mutation and Deletion in Topoisomerase IIα mRNA of Etoposide and mAMSA–resistant Cell Lines
title_sort incidence of mutation and deletion in topoisomerase iiα mrna of etoposide and mamsa–resistant cell lines
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5926608/
https://www.ncbi.nlm.nih.gov/pubmed/11676865
http://dx.doi.org/10.1111/j.1349-7006.2001.tb01069.x
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