EFFECTS OF GLYPHOSATE EXPOSURE ON NON- TUMOR HUMAN PROSTATIC CELLS.

and crops; it reaches animals, including humans, through the food chain, posing serious risks to their health. Gly is considered a potential endocrine disruptor (EDC), able of altering estrogen homeostasis by modifying the expression/localization of estrogen receptors (ERα, ERβ)1. This study aims to verify the action of Gly on the PNT1A cell line, a non-tumor human prostate epithelial cell line. Cells were treated with different concentrations of Gly at dif- ferent time intervals. Cell viability and toxicity were assessed using MTT and LDH assays; cell viability decreased as the Gly concentration increased, while cell toxicity increased. The tests allowed us to establish two doses of Gly to be used for further investigations (3.5×10-4 M and 3.5×10-3 M). Comet assay and Western blot showed that Gly-induced cell death occurred through apoptosis. Indeed, cells treated for 24 h showed a higher rate of DNA fragmentation and increased levels of the pro-apoptotic pro- teins Bax and Bak and the concomitant decrease in the anti-apop- totic protein Bcl-2 and inactivated Caspase 3; the changes were more evident in cells treated with the higher dose of Gly. Apoptosis could be due to alterations in mitochondrial metabolism, therefore the efficiency of mitochondria was studied using the Seahorse analyser and the Cell Mito Stress Test kit. Impaired mitochondrial function was observed in Gly-treated cells. It is likely that Gly-treated cells with are struggling and try- ing to compensate by working at high efficiency, as confirmed by the marked reduction in mitochondrial proton leakage and spare respiratory capacity, both indicative of the cells inability to adapt bioenergetically in response to conditions of stress. Finally, the results of immunofluorescence analysis demonstrated that Gly acted as an EDC resulting in the activation and nuclear transloca- tion of both ERs; the latter occurred regardless of dose, faster than the specific hormone, and persisted throughout treatment. In con- clusion, the results collected show that in non-tumor prostate cells the herbicide causes cell apoptosis, mitochondria dysfunction and activation of ERs.