Reactive nitrogen oxygen species metabolize N-acetylbenzidine

A close association has been reported for certain types of cancers influenc ed by aromatic amines and infection/inflammation. Reactive nitric oxygen sp ecies (RNOS), components of the inflammatory response, are bactericidal and tumoricidal, and contribute to the deleterious effects attributed to infla mmation on normal tissues. This study assessed the possible transformation of the aromatic amine N-acetylbenzidine (ABZ) by RNOS. RNOS were generated by various conditions to react with ABZ, and samples were evaluated by HPLC . Conditions which generate nitrogen dioxide radical (NO2- + myeloperoxidas e + H2O2, ONOO-, and NO2- + HOCl) produced primarily a single new product t ermed 3'-nitro-ABZ. The myeloperoxidase-catalyzed reaction with 0.3 mM NO2- was completely inhibited by 1 mM cyanide, and not effected by 100 mM chlor ide with or without 1 mM taurine. In contrast, conditions which generate N2 O3, such as spermine NONOate, did not produce 3'-nitro-ABZ, but rather two compounds termed 4'-OH-AABP and AABP. H-1 NMR and mass spectrometry identif ied 3'-nitro-ABZ as 3'-nitro-N-acetylbenzidine, 4'-OH-AABP as 4'-OH-4-acety laminobiphenyl, and AABP as 4-acetylaminobiphenyl. Human polymorphonuclear neutrophils incubated with [H-3]-ABZ and stimulated with beta -phorbol 12-m yristate 13-acetate produced 3'-nitro-ABZ in the presence of NO2- (0.1-1 mM ). Neutrophil 3'-nitro-ABZ formation was verified by mass spectrometry and was consistent with myeloperoxidase oxidation of NO2-. The results demonstr ate that ABZ forms unique products in the presence of nitrosating and nitra ting RNOS, which could influence the carcinogenic process and serve as biom arkers for these reactive species.