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Tobacco-related-compound-induced Nitrosative Stress Injury in the Hamster Cheek Pouch

Divisions of Oral & Maxillofacial Radiology and ¹ Oral & Maxillofacial Pathology, Department of Dentistry, Faculty of Medicine and Dentistry, University of Alberta, DPC 2085, Edmonton, AB T6G 2N8, Canada;

^* corresponding author, ernest.lam{at}ualberta.ca

The nitric oxide radical (^•NO) released from tobacco-related compounds induces DNA damage, protein modifications, and cellular toxicity through the formation of peroxynitrite (ONOO^–), the reaction product of ^•NO and the oxygen radical, superoxide. We hypothesize that tobacco-related compounds are cytotoxic and induce quantifiable DNA single-strand breaks in immortalized hamster cheek pouch (POII) cells, and that an amino acid marker of ONOO^– injury, namely, 3-nitrotyrosine (3-NT), is detectable in hamster cheek pouch tissues chronically exposed to these compounds. We observed a dose-dependent decrease in POII cell viability with increasing tobacco-related compound concentrations, as well as a dose-dependent increase in DNA strand breaks. Semi-quantitative immunohistochemistry showed intense 3-NT immunoreactivity in hamster tissues treated with tobacco-related compounds compared with controls (p < 0.005). Our results suggest that tobacco-related compounds, including nicotine, are genotoxic, and that 3-NT is a quantifiable marker of ONOO^– damage in intact hamster cheek pouch tissues.

KEY WORDS: tobacco-related compounds • protein nitrosation • DNA damage • oral mucosa