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A modified host-cell reactivation assay to measure repair of alkylating DNA damage for assessing risk of lung adenocarcinoma.

Publication ,  Journal Article
Wang, L; Wei, Q; Shi, Q; Guo, Z; Qiao, Y; Spitz, MR
Published in: Carcinogenesis
July 2007

The nicotine-derived nitrosamine 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) induces lung adenocarcinoma through formation of DNA adducts. Our previous research on susceptibility to tobacco-induced carcinogenesis focused on benzo[a]pyrene diol epoxide (BPDE) as the in vitro mutagen for phenotype measurements of DNA repair capacity (DRC) in mammalian cells. Here, we present a modified host-cell reactivation (HCR) assay to measure lymphocytic DRC for alkylating DNA damage as is induced by the tobacco-specific nitrosamine, NNK. We substituted dimethyl sulfate (DMS) to create alkylating damage in pCMVluc plasmid DNA and established the damage-repair dose-response curves in both normal and nucleotide excision repair-deficient lymphoblastoid cell lines and in phytohemagglutinin (PHA)-stimulated primary lymphocytes. We then successfully measured the DRC in PHA-stimulated lymphocytes from 48 patients with lung adenocarcinoma and 45 cancer-free controls and tested our hypothesis that lower DRC for alkylating damage is associated with an increased risk of lung adenocarcinoma. The cases exhibited a lower mean DRC than did the controls. A >3-fold increased risk (odds ratio = 3.21; 95% confidence interval = 1.25-8.21) was found for those with DRC levels below the control median. There was no correlation between the DRC measured with this DMS-HCR assay and that from the parallel BPDE-HCR assay. Interestingly, risk increased to >10-fold for those with sub-optimal DRC measured by both DMS- and BPDE-HCR assays. We conclude that variability in DRC is a risk factor for lung cancer and our results provide proof of principle for a new assay that can assess DRC for NNK-induced DNA damage.

Duke Scholars

Published In

Carcinogenesis

DOI

ISSN

0143-3334

Publication Date

July 2007

Volume

28

Issue

7

Start / End Page

1430 / 1436

Location

England

Related Subject Headings

  • Sulfuric Acid Esters
  • Risk Factors
  • Plasmids
  • Pilot Projects
  • Phytohemagglutinins
  • Oncology & Carcinogenesis
  • Nitrosamines
  • Middle Aged
  • Male
  • Lymphocytes
 

Citation

APA
Chicago
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MLA
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Wang, L., Wei, Q., Shi, Q., Guo, Z., Qiao, Y., & Spitz, M. R. (2007). A modified host-cell reactivation assay to measure repair of alkylating DNA damage for assessing risk of lung adenocarcinoma. Carcinogenesis, 28(7), 1430–1436. https://doi.org/10.1093/carcin/bgm029
Wang, Luo, Qingyi Wei, Qiuling Shi, Zhaosheng Guo, Yawei Qiao, and Margaret R. Spitz. “A modified host-cell reactivation assay to measure repair of alkylating DNA damage for assessing risk of lung adenocarcinoma.Carcinogenesis 28, no. 7 (July 2007): 1430–36. https://doi.org/10.1093/carcin/bgm029.
Wang L, Wei Q, Shi Q, Guo Z, Qiao Y, Spitz MR. A modified host-cell reactivation assay to measure repair of alkylating DNA damage for assessing risk of lung adenocarcinoma. Carcinogenesis. 2007 Jul;28(7):1430–6.
Wang, Luo, et al. “A modified host-cell reactivation assay to measure repair of alkylating DNA damage for assessing risk of lung adenocarcinoma.Carcinogenesis, vol. 28, no. 7, July 2007, pp. 1430–36. Pubmed, doi:10.1093/carcin/bgm029.
Wang L, Wei Q, Shi Q, Guo Z, Qiao Y, Spitz MR. A modified host-cell reactivation assay to measure repair of alkylating DNA damage for assessing risk of lung adenocarcinoma. Carcinogenesis. 2007 Jul;28(7):1430–1436.
Journal cover image

Published In

Carcinogenesis

DOI

ISSN

0143-3334

Publication Date

July 2007

Volume

28

Issue

7

Start / End Page

1430 / 1436

Location

England

Related Subject Headings

  • Sulfuric Acid Esters
  • Risk Factors
  • Plasmids
  • Pilot Projects
  • Phytohemagglutinins
  • Oncology & Carcinogenesis
  • Nitrosamines
  • Middle Aged
  • Male
  • Lymphocytes