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Removal of benzo(a)pyrene diol epoxide (BPDE)-DNA adducts as a measure of DNA repair capacity in lymphoblastoid cell lines from sisters discordant for breast cancer.
Environ Mol Mutagen. 2002; 40(2):93-100.EM

Abstract

The mutagen sensitivity assay is one of the approaches used to investigate individual DNA repair capacity. This method is based on the premise that after in vitro treatment with a test mutagen, DNA from subjects with defective repair will be more damaged than DNA from those with an efficient repair system. However, very little is known about unmeasured processes that occur between cell treatment and final assessment of DNA damage. To develop a more precise assay, we modified the traditional mutagen sensitivity assay to also include measurement of DNA damage after culturing cells in the absence of mutagen. First, we treated apparently normal and xeroderma pigmentosum lymphoblastoid cell lines with various doses of benzo(a)pyrene diol epoxide (BPDE) and harvested cells at different time points. A polyclonal antiserum against BPDE-DNA was used to quantitate levels of adducts by immunoslot-blot and immunohistochemistry. Selected conditions included treatment with 10 microM BPDE, a 4-hr culture in mutagen-free medium, and immunohistochemical measurement of BPDE-DNA adducts. The method was then applied in a pilot study to 50 lymphoblastoid lines from sisters discordant for breast cancer. There was no significant difference between cases and controls in the level of BPDE-DNA adducts in lymphoblasts harvested immediately after BPDE treatment. However, after a 4-hr culture in mutagen-free medium, the level of adducts was significantly higher (P = 0.006) among cases than in controls. There was a two-fold increase in mean adduct removal in lines from nonaffected as compared to affected sisters (44% and 22% decrease, respectively). DNA repair capacity was predictive of case status (P = 0.04) in logistic regression analysis. This method, which can be easily applied to large numbers of samples, should be useful in studies to investigate the role of DNA repair in cancer risk.

Authors+Show Affiliations

Motykiewicz G
Department of Environmental Health Sciences, Mailman School of Public Health, Columbia University, New York, New York 10032, USA. gm54@columbia.edu
Faraglia B
No affiliation info available
Wang LW
No affiliation info available
Terry MB
No affiliation info available
Senie RT
No affiliation info available
Santella RM
No affiliation info available

MeSH

7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxideAdultAge of OnsetBreast NeoplasmsCell LineCulture Media, Serum-FreeDNA AdductsDNA RepairFemaleHumansLymphocytesMiddle AgedMutagensNuclear Family

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

12203401

Citation

Motykiewicz, Grazyna, et al. "Removal of Benzo(a)pyrene Diol Epoxide (BPDE)-DNA Adducts as a Measure of DNA Repair Capacity in Lymphoblastoid Cell Lines From Sisters Discordant for Breast Cancer." Environmental and Molecular Mutagenesis, vol. 40, no. 2, 2002, pp. 93-100.
Motykiewicz G, Faraglia B, Wang LW, et al. Removal of benzo(a)pyrene diol epoxide (BPDE)-DNA adducts as a measure of DNA repair capacity in lymphoblastoid cell lines from sisters discordant for breast cancer. Environ Mol Mutagen. 2002;40(2):93-100.
Motykiewicz, G., Faraglia, B., Wang, L. W., Terry, M. B., Senie, R. T., & Santella, R. M. (2002). Removal of benzo(a)pyrene diol epoxide (BPDE)-DNA adducts as a measure of DNA repair capacity in lymphoblastoid cell lines from sisters discordant for breast cancer. Environmental and Molecular Mutagenesis, 40(2), 93-100.
Motykiewicz G, et al. Removal of Benzo(a)pyrene Diol Epoxide (BPDE)-DNA Adducts as a Measure of DNA Repair Capacity in Lymphoblastoid Cell Lines From Sisters Discordant for Breast Cancer. Environ Mol Mutagen. 2002;40(2):93-100. PubMed PMID: 12203401.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Removal of benzo(a)pyrene diol epoxide (BPDE)-DNA adducts as a measure of DNA repair capacity in lymphoblastoid cell lines from sisters discordant for breast cancer. AU - Motykiewicz,Grazyna, AU - Faraglia,Beatrice, AU - Wang,Lian Wen, AU - Terry,Mary Beth, AU - Senie,Ruby T, AU - Santella,Regina M, PY - 2002/8/31/pubmed PY - 2002/10/9/medline PY - 2002/8/31/entrez SP - 93 EP - 100 JF - Environmental and molecular mutagenesis JO - Environ Mol Mutagen VL - 40 IS - 2 N2 - The mutagen sensitivity assay is one of the approaches used to investigate individual DNA repair capacity. This method is based on the premise that after in vitro treatment with a test mutagen, DNA from subjects with defective repair will be more damaged than DNA from those with an efficient repair system. However, very little is known about unmeasured processes that occur between cell treatment and final assessment of DNA damage. To develop a more precise assay, we modified the traditional mutagen sensitivity assay to also include measurement of DNA damage after culturing cells in the absence of mutagen. First, we treated apparently normal and xeroderma pigmentosum lymphoblastoid cell lines with various doses of benzo(a)pyrene diol epoxide (BPDE) and harvested cells at different time points. A polyclonal antiserum against BPDE-DNA was used to quantitate levels of adducts by immunoslot-blot and immunohistochemistry. Selected conditions included treatment with 10 microM BPDE, a 4-hr culture in mutagen-free medium, and immunohistochemical measurement of BPDE-DNA adducts. The method was then applied in a pilot study to 50 lymphoblastoid lines from sisters discordant for breast cancer. There was no significant difference between cases and controls in the level of BPDE-DNA adducts in lymphoblasts harvested immediately after BPDE treatment. However, after a 4-hr culture in mutagen-free medium, the level of adducts was significantly higher (P = 0.006) among cases than in controls. There was a two-fold increase in mean adduct removal in lines from nonaffected as compared to affected sisters (44% and 22% decrease, respectively). DNA repair capacity was predictive of case status (P = 0.04) in logistic regression analysis. This method, which can be easily applied to large numbers of samples, should be useful in studies to investigate the role of DNA repair in cancer risk. SN - 0893-6692 UR - https://www.unboundmedicine.com/medline/citation/12203401/Removal_of_benzo_a_pyrene_diol_epoxide__BPDE__DNA_adducts_as_a_measure_of_DNA_repair_capacity_in_lymphoblastoid_cell_lines_from_sisters_discordant_for_breast_cancer_ DB - PRIME DP - Unbound Medicine ER -