Metabolism-mediated Ochratoxin A genotoxicity in the single-cell gel electrophoresis (Comet) assay

The mycotoxin Ochratoxin A (OTA) is a contaminant of food and feed commodities in many countries. Long-term exposure to OTA in humans has been associated with an increased incidence of a progressive nephropathy (BEN). Bio-activation of OTA has been implicated in the OTA-mediated toxicity, although inconsistent results have been reported. The aim of the present study was to investigate the genotoxic potency of OTA and its metabolites in NIH/3T3 cells stably expressing the human cytochrome P450 iso-enzymes CYP2C9 and CYP3A4, by using the single-cell gel electrophoresis (SCGE/Comet) assay, which detects single strand DNA breaks. The obtained results confirm the hypothesis that biotransformation processes mediate OTA toxicity, and differences in response were observed in CYP2C9-hOR and CYP3A4-hOR expressing cells, respectively. Results showed that biotransformation of OTA increased the genotoxicity. Measurement of reactive oxygen species (ROS) production showed that the OTA-induced ROS production corresponded to the OTA-induced genotoxicity in the used NIH/3T3 cell lines.