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An immunoperoxidase method, using a monoclonal antibody which recognizes 4-aminobiphenyl (4-ABP)-DNA adducts, was developed for the detection and quantitation of DNA damage in bladder tissue and applied to stored paraffin blocks of transurethral resection specimens of 46 patients with T1 bladder cancer. Mean relative staining intensity for 4-ABP-DNA adducts was significantly higher in current smokers (275 +/- 81, n = 24) compared to nonsmokers (113 +/- 71, n = 22) (P &lt; 0.0001). There was a linear relationship between mean levels of relative staining and number of cigarettes smoked with lower levels in the 1-19 cig/day group (205 +/- 30, n = 5), compared to the 20-40 (289 +/- 40, n = 7) and the &gt;40 cig/day group (351 +/- 57, n = 3)(P &lt; 0.001). Nuclear overexpression of p53, analyzed by immunoperoxidase staining, was observed in 27 (59%) of the 45 stage T1 tumors analyzed. There was a significant correlation between p53 overexpression and recurrence of disease (odds ratio = 12.3, P &lt; 0.01). Nuclear staining of p53 was also correlated with smoking status, cig/day and 4-ABP-DNA adducts. This work demonstrates that the immunohistochemical method has sufficient sensitivity for detection of 4-ABP-DNA adducts in human bladder samples. The method has several advantages including small sample size, the possibility of retrospective analysis of stored paraffin blocks, the ability to analyze binding in specific cell types, and a relatively low cost.

Immunohistochemical quantitation of 4-aminobiphenyl-DNA adducts and p53 nuclear overexpression in T1 bladder cancer of smokers and nonsmokers