Cytotoxic impact of DNA single vs double strand breaks in oxidatively injured cells

Hydrogen peroxide is a potent inducer of DNA single strand breaks (SSBs) in cultured mammalian cells. These lesions, however, are efficiently repaired and do not appear to mediate the cytotoxic response. This inference is based on the observations that a) inhibiting the rate of SSB-removal does not result in an increased cytotoxicity; b) using different experimental conditions it is possible to dissociate the formation of DNA SSBs from the cytotoxic response; c) the induction/loss of the oxidant-resistant phenotype in cell variants characterized by different levels of resistance to the lethal effect of the oxidant does not correlate with resistance to DNA SSB-induction; d) a much larger accumulation of DNA SSBs can be observed following treatment with H2O2 at 4 degrees C, as compared to 37 degrees C, although the opposite is true in terms of cytotoxicity. In the presence of micromolar levels of L-Histidine, H2O2 also induces DNA double strand breaks (DSBs), a type of lesion which we suggest may mediate the lethal event. This conclusion finds experimental support in the following observations: a) DNA DSBs are generated at survival-range concentrations, and a linear correlation exists between the level of this lesion and cytotoxicity; b) this correlation curve overlaps with the curves generated under similar experimental conditions using different cell lines with different sensitivity to the oxidant alone, or different clones derived from the same cell line, some of which showed a high degree of resistance to H2O2. Finally, the formation of DNA DSBs appears to enhance both apoptotic and necrotic cell deat