32P-Postlabelling of aromatic DNA adducts in white blood cells and alveolar macrophages of smokers: saturation at high exposures

Abstract

DNA adducts may serve as a molecular dosimeter of exposure to cigarette smoke-associated carcinogens such as polycyclic aromatic hydrocarbons (PAH). Target tissues for cigarette smoke-induced carcinogenesis are rarely accessible; therefore, peripheral blood cells or cells obtained by bronchoalveolar lavage (BAL) may be used as surrogate sources of exposed DNA. However, the relationship between cigarette smoke exposure and aromatic–DNA adducts in white blood cells and BAL cells is still unclear. In this study, we examined DNA adduct formation in lymphocytes and BAL cells in several populations of smoking individuals by means of 32P-postlabelling. Significant correlations between the amount of cigarettes smoked per day and the level of aromatic–DNA adducts were found in lymphocytes. In BAL cells, DNA adduct levels were associated with age ( p=0.05) and gender ( p=0.10) after adjustment for smoking behaviour. Adduct formation levelled off at higher exposure levels, suggesting less efficient adduct formation; decreases in the formation of adducts per unit of exposure were found in lymphocytes ( r s=−0.80, p<0.001) and BAL cells ( r s=−0.72, p<0.001). To assess intra-individual variation in adduct levels at constant smoking behaviour, sampling was repeated after a period of 2 and 6 months. In lymphocytes, repeated measurements with an interval of 2 months were highly correlated ( r=0.84, p=0.009, n=8), whereas repeated measurements with an interval of 6 months showed no correlation ( r=0.30, p=0.27, n=16). Repeated measurements in BAL cells showed a significant correlation after 6 months ( r=0.68, p=0.03, n=10). Furthermore, in a group of occupationally exposed aluminium workers, adduct levels in total white blood cells were correlated with the average concentrations of PAH in the ambient air of workers who smoked cigarettes, whereas in non-smokers, no such relationship was found. We conclude that cigarette smoking may directly or indirectly influence DNA adduct levels and saturation of DNA adduct formation may occur, leading to non-linear dose–response relationships.