Smokeless Tobacco And Dna Ladder Analysis In Human Oral Keratinocytes

The status of genomic DNA (qualitative and quantitative) during STE treatment of keratinocytes was assessed. Agarose gel electrophoresis was used to provide evidence that NHOK treated with increasing concentrations of STE (0 to 300^g/ml) for 24h exhibited DNA damage. STE induced a ladderlike pattern of endonucleolytic DNA degradation at all concentrations. The characteristic ladder pattern of internucleosomal DNA cleavage that was observed is consistent with activation of an endogenous endonuclease and is readily evident in STE-treated cells.28'45

The fragmentation pattern for DNA from cells treated with STE indicated significant DNA damage caused by STE' while DNA from the control cells remained totally unfragmented. At a concentration of 100^g STE/ml, the ladder formation was not very prominent.28 However, at higher concentrations, the loss of higher-molecular-weight genomic DNA was observed, and a concentration-dependent response occurred. Partial reversal of STE-induced oligonucleosome length DNA-laddering was observed in cells treated with the various antioxidants. Greatest protection against STE-induced DNA fragmentation was provided by grape seed proanthocyanidin extract (GSPE).28

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