Genotoxic effects of doxorubicin in cultured human lymphocytes with different glutathione S-transferase genotypes

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

Doxorubicin (Dox) is a widely used drug in oncology with a broad spectrum of interactions with various cellular components; therefore, it is likely to act through different mechanisms. In clinical practice there is inter-individual variability in cytotoxic drug response and in the occurrence of adverse reactions. Glutathione S-transferases (GSTM1, GSTT1 and GSTP1) are thought to be involved in the detoxification of endogenous and exogenous genotoxicants. The aim of this work is the assessment of a possible influence of polymorphisms in GSTs on the levels of genetic damage induced in vitro by Dox in cultured human lymphocytes. For this purpose, whole blood cultures from individuals with different genotypes for GSTM1. GSTT1 and GSTP1 were exposed to Dox and the cytokinesis-blocked micronucleus (CBMN) assay was used as the endpoint for chromosomal damage in the lymphocytes. Genotyping of GSTM1 and GSTT1 was carried out by multiplex PCR and the GSTP1-Ile105Val polymorphism was determined by PCR/RFLP. The total number of micronuclei present in 1000 binucleated cells and the frequency of micronucleated binucleated lymphocytes in the different individuals were analyzed considering the GSTM1, GSTT1 and GSTP1 genotypes. The results obtained suggest that GSTM1 and GSTT1 deletion polymorphisms do not modify significantly the genotoxic potential of Dox. However, the GSTP1 Ile105Val polymorphism was associated with an increase of micronucleated binucleated cells induced by Dox. Lymphocytes from homozygous individuals for the variant form (Val/Val) presented a significant increase in micronucleated binucleated cells (approximately 1.5-fold; p < 0.05) when compared with individuals with at least one wild-type allele. These results suggest a possible role for GSTP1 on the modulation of the genotoxicity induced by Dox, which may be considered in cancer therapy. (C) 2011 Elsevier By. All rights reserved.
Original languageEnglish
Pages (from-to)28-34
Number of pages7
JournalMutation Research-Genetic Toxicology And Environmental Mutagenesis
Volume724
Issue number1-2
DOIs
Publication statusPublished - 2011

Fingerprint

Glutathione Transferase
Doxorubicin
Genotype
Lymphocytes
Micronucleus Tests
Cytokinesis
Multiplex Polymerase Chain Reaction
Restriction Fragment Length Polymorphisms
Pharmaceutical Preparations
Alleles
Polymerase Chain Reaction
Neoplasms

Cite this

@article{a9b45d974cf54a2ebdec28dd1441ba74,
title = "Genotoxic effects of doxorubicin in cultured human lymphocytes with different glutathione S-transferase genotypes",
abstract = "Doxorubicin (Dox) is a widely used drug in oncology with a broad spectrum of interactions with various cellular components; therefore, it is likely to act through different mechanisms. In clinical practice there is inter-individual variability in cytotoxic drug response and in the occurrence of adverse reactions. Glutathione S-transferases (GSTM1, GSTT1 and GSTP1) are thought to be involved in the detoxification of endogenous and exogenous genotoxicants. The aim of this work is the assessment of a possible influence of polymorphisms in GSTs on the levels of genetic damage induced in vitro by Dox in cultured human lymphocytes. For this purpose, whole blood cultures from individuals with different genotypes for GSTM1. GSTT1 and GSTP1 were exposed to Dox and the cytokinesis-blocked micronucleus (CBMN) assay was used as the endpoint for chromosomal damage in the lymphocytes. Genotyping of GSTM1 and GSTT1 was carried out by multiplex PCR and the GSTP1-Ile105Val polymorphism was determined by PCR/RFLP. The total number of micronuclei present in 1000 binucleated cells and the frequency of micronucleated binucleated lymphocytes in the different individuals were analyzed considering the GSTM1, GSTT1 and GSTP1 genotypes. The results obtained suggest that GSTM1 and GSTT1 deletion polymorphisms do not modify significantly the genotoxic potential of Dox. However, the GSTP1 Ile105Val polymorphism was associated with an increase of micronucleated binucleated cells induced by Dox. Lymphocytes from homozygous individuals for the variant form (Val/Val) presented a significant increase in micronucleated binucleated cells (approximately 1.5-fold; p < 0.05) when compared with individuals with at least one wild-type allele. These results suggest a possible role for GSTP1 on the modulation of the genotoxicity induced by Dox, which may be considered in cancer therapy. (C) 2011 Elsevier By. All rights reserved.",
keywords = "genetic, p1, susceptibility, Genetic, breast-cancer, Doxorubicin, polymorphisms, cell-survival, Micronuclei, S-transferases, gamma-radiation, drug-resistance, risk, anthracyclines, micronuclei, Glutathione",
author = "Gaspar, {Jorge F.} and Marta Pingarilho and J. Rueff and Fernandes, {Ana Sofia}",
year = "2011",
doi = "10.1016/j.mrgentox.2011.04.013",
language = "English",
volume = "724",
pages = "28--34",
journal = "Mutation Research-Genetic Toxicology And Environmental Mutagenesis",
issn = "1383-5718",
publisher = "Elsevier Science B.V., Inc",
number = "1-2",

}

TY - JOUR

T1 - Genotoxic effects of doxorubicin in cultured human lymphocytes with different glutathione S-transferase genotypes

AU - Gaspar, Jorge F.

AU - Pingarilho, Marta

AU - Rueff, J.

AU - Fernandes, Ana Sofia

PY - 2011

Y1 - 2011

N2 - Doxorubicin (Dox) is a widely used drug in oncology with a broad spectrum of interactions with various cellular components; therefore, it is likely to act through different mechanisms. In clinical practice there is inter-individual variability in cytotoxic drug response and in the occurrence of adverse reactions. Glutathione S-transferases (GSTM1, GSTT1 and GSTP1) are thought to be involved in the detoxification of endogenous and exogenous genotoxicants. The aim of this work is the assessment of a possible influence of polymorphisms in GSTs on the levels of genetic damage induced in vitro by Dox in cultured human lymphocytes. For this purpose, whole blood cultures from individuals with different genotypes for GSTM1. GSTT1 and GSTP1 were exposed to Dox and the cytokinesis-blocked micronucleus (CBMN) assay was used as the endpoint for chromosomal damage in the lymphocytes. Genotyping of GSTM1 and GSTT1 was carried out by multiplex PCR and the GSTP1-Ile105Val polymorphism was determined by PCR/RFLP. The total number of micronuclei present in 1000 binucleated cells and the frequency of micronucleated binucleated lymphocytes in the different individuals were analyzed considering the GSTM1, GSTT1 and GSTP1 genotypes. The results obtained suggest that GSTM1 and GSTT1 deletion polymorphisms do not modify significantly the genotoxic potential of Dox. However, the GSTP1 Ile105Val polymorphism was associated with an increase of micronucleated binucleated cells induced by Dox. Lymphocytes from homozygous individuals for the variant form (Val/Val) presented a significant increase in micronucleated binucleated cells (approximately 1.5-fold; p < 0.05) when compared with individuals with at least one wild-type allele. These results suggest a possible role for GSTP1 on the modulation of the genotoxicity induced by Dox, which may be considered in cancer therapy. (C) 2011 Elsevier By. All rights reserved.

AB - Doxorubicin (Dox) is a widely used drug in oncology with a broad spectrum of interactions with various cellular components; therefore, it is likely to act through different mechanisms. In clinical practice there is inter-individual variability in cytotoxic drug response and in the occurrence of adverse reactions. Glutathione S-transferases (GSTM1, GSTT1 and GSTP1) are thought to be involved in the detoxification of endogenous and exogenous genotoxicants. The aim of this work is the assessment of a possible influence of polymorphisms in GSTs on the levels of genetic damage induced in vitro by Dox in cultured human lymphocytes. For this purpose, whole blood cultures from individuals with different genotypes for GSTM1. GSTT1 and GSTP1 were exposed to Dox and the cytokinesis-blocked micronucleus (CBMN) assay was used as the endpoint for chromosomal damage in the lymphocytes. Genotyping of GSTM1 and GSTT1 was carried out by multiplex PCR and the GSTP1-Ile105Val polymorphism was determined by PCR/RFLP. The total number of micronuclei present in 1000 binucleated cells and the frequency of micronucleated binucleated lymphocytes in the different individuals were analyzed considering the GSTM1, GSTT1 and GSTP1 genotypes. The results obtained suggest that GSTM1 and GSTT1 deletion polymorphisms do not modify significantly the genotoxic potential of Dox. However, the GSTP1 Ile105Val polymorphism was associated with an increase of micronucleated binucleated cells induced by Dox. Lymphocytes from homozygous individuals for the variant form (Val/Val) presented a significant increase in micronucleated binucleated cells (approximately 1.5-fold; p < 0.05) when compared with individuals with at least one wild-type allele. These results suggest a possible role for GSTP1 on the modulation of the genotoxicity induced by Dox, which may be considered in cancer therapy. (C) 2011 Elsevier By. All rights reserved.

KW - genetic

KW - p1

KW - susceptibility

KW - Genetic

KW - breast-cancer

KW - Doxorubicin

KW - polymorphisms

KW - cell-survival

KW - Micronuclei

KW - S-transferases

KW - gamma-radiation

KW - drug-resistance

KW - risk

KW - anthracyclines

KW - micronuclei

KW - Glutathione

U2 - 10.1016/j.mrgentox.2011.04.013

DO - 10.1016/j.mrgentox.2011.04.013

M3 - Article

VL - 724

SP - 28

EP - 34

JO - Mutation Research-Genetic Toxicology And Environmental Mutagenesis

JF - Mutation Research-Genetic Toxicology And Environmental Mutagenesis

SN - 1383-5718

IS - 1-2

ER -