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Preliminary Evaluation of DNA Damage Related with the Smoking Habit Measured by the Comet Assay in Whole Blood Cells

¹ Laboratorio de Microbiología y Bioensayos, Centro Nacional del Medio Ambiente (CENMA), Universidad de Chile, Santiago, Chile and ² Toxic Substances Research and Teaching Program, University of California, Davis, California

Requests for reprints: Manuel Ellahueñe, Centro Nacional del Medio Ambiente (CENMA), Av. Larrain 9975, La Reina, Santiago, Chile. Phone: 562-2994151; Fax: 562-2751688. E-mail: mellahuene{at}cenma.cl

The alkaline single-cell gel electrophoresis (SCGE) assay, also called the comet assay, is a rapid and simple method for the detection of DNA damage in individual cells. The objective of this study was to establish if the alkaline SCGE assay in whole blood cells gives similar results as the same method in isolated lymphocytes, because whole blood cells are simpler and more economical to use, specifically in human genotoxic biomonitoring. To validate the method, we first used mouse blood cells, because mouse is one of the most commonly used animals in genetic toxicology testing. Groups of seven CF1 male mice were given i.p. injections of relatively low doses of methyl methanesulfonate (25 mg/kg body weight), a direct acting genotoxic agent, or cyclophosphamide (50 mg/kg body weight), which requires metabolic activation. Three, 6, 8, 12, 16, 20, and 65 hours after treatment, 5 µL of blood were collected from each animal and were processed for the alkaline SCGE assay. On the basis of an analysis of tail moment, the results showed that this assay can detect DNA damage induced by both kinds of alkylating mutagens. We then did a preliminary study to assess the status of DNA damage in a young (19 to 23 years old) healthy population of male smokers (n = 6) and nonsmokers (n = 6) using the comet assay in whole blood cells. A significant difference was observed between the two groups, showing that the method is able to detect DNA damage in the smoking group despite the short time that the volunteers had actually been smoking.

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