Mutagenesis, Vol. 16, No. 1, 25-30,
January 2001
© 2001 UK Environmental Mutagen Society/Oxford University Press
Rapid screening assay for mutagen sensitivity and DNA repair capacity in human peripheral blood lymphocytes
1 Division of Toxicology and Cancer Risk Factors, German Cancer Research Center, Heidelberg, 2 Biostatistics Unit, German Cancer Research Center, Heidelberg and 3 Thoraxklinik Heidelberg-Rohrbach, Germany
Individual susceptibility to carcinogens, an important determinant of disease risk, is influenced by host factors such as the ability to repair DNA lesions. In order to identify subjects who are at high risk, we have developed a microgel electrophoresis assay for use in molecular epidemiological studies. The assay was validated in a pilot casecontrol study: Peripheral blood lymphocytes were collected from 100 patients with lung cancer and 110 control patients without cancer and from the same hospital, and stored at 80°C. After thawing, phytohaemagglutinin-stimulated cells were treated with bleomycin at 20 µg/ml for 30 min and the extent of DNA damage and DNA repair capacity were determined by microgel electrophoresis. Peripheral blood lymphocytes from patients with lung cancer were significantly more sensitive to mutagens than those from controls and showed reduced DNA repair capacity (both P < 0.001). Both endpoints were independent risk factors for smoking-related lung cancer. Repeated analysis of peripheral blood lymphocytes from the same individual demonstrated good reproducibility of the assay. Cryopreservation of the lymphocytes for
12 months did not significantly affect their sensitivity. Our standardized microgel electrophoresis assay is suitable for determining individual sensitivity to mutagens and DNA repair capacity: it is sensitive and faster than cytogenetic assays, and can be applied to native and cryopreserved peripheral blood lymphocytes.
4 To whom correspondence should be addressed. E-mail: p.schmezer{at}dkfz-heidelberg.de
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