Smokeless Tobacco And Dna Fragmentation In Human Oral Keratinocytes

DNA fragmentation is a biological hallmark of oxidative DNA and cellular injury.44 The cyto-toxicity of STE was examined by determining DNA fragmentation (DNA damage)

24h after treatment of normal human oral keratinocytes (NHOK) in culture with 100, 250, and 350^g/ml of STE. STE induced significant DNA damage, as measured by DNA fragmentation.40 Effects of STE treatment on keratinocytes were reported as percent of fragmentation relative to untreated cells. DNA fragmentation in samples is expressed as percent of total DNA appearing in the supernatant fractions.40'44 The absorbencies were measured spectrometrically at 600 nm. A concentration-dependent increase in DNA fragmentation was observed with increases of 1.5-, 1.6-, and 2.9-fold following exposure of the cells to the three concentrations of STE, respectively. Pretreatment of the NHOK with vitamin C (75^M), vitamin E (75^M), a combination of vitamins C plus E (75^M each), and GSPE (100^g/ml) for 4h decreased STE (300^g/ml)-induced DNA fragmentation by 9, 12, 20, and 36%, respectively, as compared with the STE-treated cells. The results demonstrate that increased production of ROS with STE treatment causes oxidative DNA damage and cell death.

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