Possible involvement of oxidative stress in potassium bromate-induced genotoxicity in human HepG2 cells

Abstract

Potassium bromate (KBrO(3), PB) is a by-product of ozone used as disinfectant in drinking water. And PB is also a widely used food additive. However, there is little known about its adverse effects, in particular those related to its genotoxicity in humans. The aim of this study was to investigate the genotoxic effects of PB and the underlying mechanisms, using human hepatoma cell line, HepG2. Exposure of the cells to PB caused a significant increase of DNA migration in single cell gel electrophoresis (SCGE) assay and micronuclei (MN) frequencies in micronucleus test (MNT) at all tested concentrations (1.56-12.5 mM and 0.12-1 mM), which suggested that PB-mediated DNA strand breaks and chromosome damage. To indicate the role of antioxidant in those effects, DNA migration was monitored by pre-treatment with hydroxytyrosol (HT) as an antioxidant in SCGE assay. It was found that DNA migration with pre-treatment of HT was dramatically decreased. To elucidate the genotoxicity mechanisms, the study monitored the levels of reactive oxygen species (ROS), glutathione (GSH) and 8-hydroxydeoxyguanosine (8-OHdG). PB was shown to induce ROS production (12.5 mM), GSH depletion (1.56-12.5 mM) and 8-OHdG formation (6.25-12.5 mM) in HepG2 cells. Moreover, lysosomal membrane stability and mitochondrial membrane potential were further studied for the mechanisms of PB-induced genotoxicity. A significant increase was found in the range of 6.25-12.5 mM in lysosomal membrane stability assay. However, under these PB concentrations, we were not able to detect the changes of mitochondrial membrane potential. These results suggest that PB exerts oxidative stress and genotoxic effects in HepG2 cells, possibly through the mechanisms of lysosomal damage, an earlier event preceding the oxidative DNA damage.