Cylindrospermopsin induced DNA damage and alteration in the expression of genes involved in the response to DNA damage, apoptosis and oxidative stress

Abstract

Cylindrospermopsin (CYN), a potent cyanobacterial cytototoxin produced by certain freshwater cyanobacteria, is regularly found in water supplies in many parts of the world, and has been associated with the intoxication of humans and livestock. The few genotoxicity studies available indicate that CYN is genotoxic, generally implying that it is pro-genotoxic. In human peripheral blood lymphocytes (HPBLs) CYN (0, 0.05, 0.1 and 0.5 μg/ml) induced the formation of DNA single strand breaks, applying the comet assay. Time and dose dependent significant increase in the frequency of micronuclei and nuclear buds was observed after the exposure of HPBLs to CYN, while there was only slight increase in the number of nucleoplasmic bridges. For the first time the modulation of gene expression in HPBLs was studied after the exposure to CYN (0.5 μg/ml), using the quantitative real-time PCR. The genes presumably involved in CYN metabolism (CYP1A1 and CYP1A2) were up-regulated after the exposure. CYN induced changes in the mRNA expression of P53 and its downstream regulated DNA damage responsive genes MDM2, GADD45α and apoptosis genes, BCL-2 and BAX, as well as oxidative stress responsive genes (GPX1, SOD1, GSR, GCLC), while no changes in the expression of genes CDKN1A and CAT were observed. These results provide strong evidence that CYN should be considered as genotoxic and that lymphocytes can also be a target of cylindrospermopsin induced genotoxicity.