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Rapid method for determination of DNA repair capacity in human peripheral blood lymphocytes amongst smokers

BACKGROUND: DNA repair capacity is an important determinant of susceptibility to cancer. The hOGG1 enzyme is crucial for repairing the 8-oxoguanine lesion that occurs either as a byproduct of oxidative metabolism or as a result of exogenous sources such as exposure to cigarette smoke. It has been pr...

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Autores principales: El-Zein, Randa A, Monroy, Claudia M, Cortes, Andrea, Spitz, Margaret R, Greisinger, Anthony, Etzel, Carol J
Formato: Texto
Lenguaje:English
Publicado: BioMed Central 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2933626/
https://www.ncbi.nlm.nih.gov/pubmed/20718982
http://dx.doi.org/10.1186/1471-2407-10-439
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author El-Zein, Randa A
Monroy, Claudia M
Cortes, Andrea
Spitz, Margaret R
Greisinger, Anthony
Etzel, Carol J
author_facet El-Zein, Randa A
Monroy, Claudia M
Cortes, Andrea
Spitz, Margaret R
Greisinger, Anthony
Etzel, Carol J
author_sort El-Zein, Randa A
collection PubMed
description BACKGROUND: DNA repair capacity is an important determinant of susceptibility to cancer. The hOGG1 enzyme is crucial for repairing the 8-oxoguanine lesion that occurs either as a byproduct of oxidative metabolism or as a result of exogenous sources such as exposure to cigarette smoke. It has been previously reported that smokers with low hOGG1 activity had significantly higher risk of developing lung cancer as compared to smokers with high hOGG1 activity. METHODS: In the current study we elucidate the association between plasma levels of 8-OHdG and the OGG1 repair capacity. We used the commercially available 8-OHdG ELISA (enzyme-linked immunosorbent assay), the Comet assay/FLARE hOGG1 (Fragment Length Analysis by Repair Enzymes) assay for quantification of the levels of 8-OHdG and measured the constitutive, induced and unrepaired residual damage, respectively. We compared the DNA repair capacity in peripheral blood lymphocytes following H(2)O(2 )exposure in 30 lung cancer patients, 30 non-, 30 former and 30 current smoker controls matched by age and gender. RESULTS: Our results show that lung cancer cases and current smoker controls have similar levels of 8-OHdG lesions that are significantly higher compared to the non-smokers controls. However, lung cancer cases showed significantly poorer repair capacity compared to all controls tested, including the current smokers controls. After adjustment for age, gender and family history of smoking-related cancer using linear regression, we observed a 5-fold increase in risk of lung cancer associated with high levels of residual damage/reduced repair capacity. Reduced OGG1 activity could be expected to be a risk factor in other smoking-related cancers. CONCLUSION: Our study shows that the Comet/FLARE assay is a relatively rapid and useful method for determination of DNA repair capacity. Using this assay we could identify individuals with high levels of residual damage and hence poor repair capacity who would be good candidates for intensive follow-up and screening.
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spelling pubmed-29336262010-09-08 Rapid method for determination of DNA repair capacity in human peripheral blood lymphocytes amongst smokers El-Zein, Randa A Monroy, Claudia M Cortes, Andrea Spitz, Margaret R Greisinger, Anthony Etzel, Carol J BMC Cancer Research Article BACKGROUND: DNA repair capacity is an important determinant of susceptibility to cancer. The hOGG1 enzyme is crucial for repairing the 8-oxoguanine lesion that occurs either as a byproduct of oxidative metabolism or as a result of exogenous sources such as exposure to cigarette smoke. It has been previously reported that smokers with low hOGG1 activity had significantly higher risk of developing lung cancer as compared to smokers with high hOGG1 activity. METHODS: In the current study we elucidate the association between plasma levels of 8-OHdG and the OGG1 repair capacity. We used the commercially available 8-OHdG ELISA (enzyme-linked immunosorbent assay), the Comet assay/FLARE hOGG1 (Fragment Length Analysis by Repair Enzymes) assay for quantification of the levels of 8-OHdG and measured the constitutive, induced and unrepaired residual damage, respectively. We compared the DNA repair capacity in peripheral blood lymphocytes following H(2)O(2 )exposure in 30 lung cancer patients, 30 non-, 30 former and 30 current smoker controls matched by age and gender. RESULTS: Our results show that lung cancer cases and current smoker controls have similar levels of 8-OHdG lesions that are significantly higher compared to the non-smokers controls. However, lung cancer cases showed significantly poorer repair capacity compared to all controls tested, including the current smokers controls. After adjustment for age, gender and family history of smoking-related cancer using linear regression, we observed a 5-fold increase in risk of lung cancer associated with high levels of residual damage/reduced repair capacity. Reduced OGG1 activity could be expected to be a risk factor in other smoking-related cancers. CONCLUSION: Our study shows that the Comet/FLARE assay is a relatively rapid and useful method for determination of DNA repair capacity. Using this assay we could identify individuals with high levels of residual damage and hence poor repair capacity who would be good candidates for intensive follow-up and screening. BioMed Central 2010-08-18 /pmc/articles/PMC2933626/ /pubmed/20718982 http://dx.doi.org/10.1186/1471-2407-10-439 Text en Copyright ©2010 El-Zein et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
El-Zein, Randa A
Monroy, Claudia M
Cortes, Andrea
Spitz, Margaret R
Greisinger, Anthony
Etzel, Carol J
Rapid method for determination of DNA repair capacity in human peripheral blood lymphocytes amongst smokers
title Rapid method for determination of DNA repair capacity in human peripheral blood lymphocytes amongst smokers
title_full Rapid method for determination of DNA repair capacity in human peripheral blood lymphocytes amongst smokers
title_fullStr Rapid method for determination of DNA repair capacity in human peripheral blood lymphocytes amongst smokers
title_full_unstemmed Rapid method for determination of DNA repair capacity in human peripheral blood lymphocytes amongst smokers
title_short Rapid method for determination of DNA repair capacity in human peripheral blood lymphocytes amongst smokers
title_sort rapid method for determination of dna repair capacity in human peripheral blood lymphocytes amongst smokers
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2933626/
https://www.ncbi.nlm.nih.gov/pubmed/20718982
http://dx.doi.org/10.1186/1471-2407-10-439
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