Binding and uptake of N-nitrosonornicotine by oral epithelial cells

N-nitrosonornicotine (NNN), a significant nitrosamine component of tobacco binds to and is taken up by cells prior to its activation by intracellular enzymes. A variety of factors may affect the binding of substances to a cell including the cell type, medium composition and the cell membrane lipid composition. This study examines the binding and uptake of NNN to various cell types and relates these to the cells' lipid composition. Hamster buccal pouch keratinocytes were exposed to NNN and the amount of bound NNN determined. This was compared with cells pretreated with 12-0-tetradecanoylphorbol-13-acetate (TPA). While the keratinocytes bound significant amounts of NNN, this binding was not specific. Pretreatment of the cells with TPA significantly increased the amount of NNN bound. Cultures of human gingival fibroblasts and a human liver cell line also bound NNN, in quantities greater than that bound to the keratinocytes. The TPA caused changes in long chain unsaturated fatty acids although major changes in lipid metabolism were not detected. The fatty acid composition of the liver cells more closely resembled that of the TPA treated keratinocytes than the untreated ones. The data suggest that NNN binds to the cells non-specifically and the binding may be related to the amount of long-chain, unsaturated fatty acids present.