Metabolism-mediated cytotoxicity of ochratoxin A

Ochratoxin A (OTA) is produced by various strains of Aspergillus and Penicillium and is a common contaminant of food commodities. OTA is metabolised by cytochrome P450 (CYP450) enzymes resulting in hydroxylated metabolites, 4R-OH-OTA and 4S-OH-OTA, and possibly in other minor metabolites including OTA-quinones. However, until now conflicting data have been presented regarding the role of biotransformation products in the adverse effects of OTA. Hence, the aim of this study was to further assess the metabolism-mediated cytotoxicity of OTA in an in vitro model encompassing NIH/3T3 cells, stably expressing the human CYP450 enzymes CYP2C9 and CYP3A4, respectively. In addition, modulation of the cellular glutathione (GSH) content was used to identify a role of GSH in OTA-induced cytotoxicity. Following exposure to OTA, cells expressing CYP2C9 showed a significant reduction in neutral red (NR) uptake but not in Alamar blue (AB) reduction, as compared to the control LNCX cells which do not express CYP450 enzymes. CYP3A4-expressing cells showed no difference in viability from control LNCX cells. When pretreated with L-buthionine S,R-sulphoximine (BSO) to deplete GSH, CYP2C9-expressing cells showed also a loss of cell viability as compared to LNCX cells, although to a lesser extent as compared to non-depleted CYP2C9-expressing cells. Data presented in this study support previous findings, indicating that different biotransformation pathways contribute to the cytotoxicity induced by OTA. (C) 2003 Elsevier Ltd. All rights reserved.