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MULTIPLE DNA ADDUCTS IN LYMPHOCYTES OF SMOKERS AND NONSMOKERS DETERMINED BY 32P-POSTLABELING ANALYSIS
Citation:
Jahnke, G., C. Thompson, M. Walker, J. Gallagher, G. Lucier, AND R. DiAugustine. MULTIPLE DNA ADDUCTS IN LYMPHOCYTES OF SMOKERS AND NONSMOKERS DETERMINED BY 32P-POSTLABELING ANALYSIS. U.S. Environmental Protection Agency, Washington, D.C., EPA/600/J-90/177 (NTIS PB91116897).
Description:
Identification of DNA adducts in peripheral lymphocytes could serve as a means of monitoring human exposure to potential genotoxic agents. n this study, DNA from peripheral lymphocytes of smokers and nonsmokers was examined for adducts by the P1 nuclease 32P-postlabeling technique. hin layer chromatography (TLC) maps from both groups revealed multiple DNA adducts which ranged from no adducts for one individual to six adducts for a different individual. he total DNA adduct concentrations were approximately one adduct in 10 7-10 8 normal nucleotides. omparison of the adduct TLC profiles revealed individual variation in both pattern and level of DNA adducts. he type and amount of adduct was not influenced by smoking history and remained unchanged in four out of six subjects who were resampled after a one month interval. ne adduct detected in lymphocyte DNA co-migrated on TLC with an adduct derived by in incubation of lymphocytes with benzo(a)pyrene [B(a)P]. he capacity of lymphocytes to form B(a)P-derived DNA adducts after a 72 h incubation ith 10-6M [3H]-B(a)P was measured by both HPLC and 32P-postlabeling analysis. he 3H-nucleoside alues were consistent ith values obtained by 32P-postlabeling of the same sample (correlation coefficient of 0.88). o relationship was apparent between the capacity of lymphocytes to form a [3H]-B(a)P-derived adduct in vitro and the concentration of this adduct, or total adducts present in untreated lymphocytes. hese results suggest that multiple DNA adducts are present in lymphocytes from nonsmokers as well as smokers, although the profile and extent of these adducts can vary among individuals. he relationship of the lymphocyte DNA adducts detected in this study to human cancer susceptibility remains to be determined.