The investigation of cytotoxic effect of sodium fluoride (NAF) in renal cell line

This study was planned to investigate the possible roles of apoptotic, autophagic and necrotic mechanisms for cytotoxicity by NaF administration in the NRK-52 renal cell line, at different doses and times. Cells were replicated in regular passages in vitro conditions. The IC50 value of NaF was determined with MTT. Cell samples were taken at 3, 12 and 24 hours from samples prepared according to the IC50 values of the cells. For biochemical assays, cells were harvested by trypsinization and lysed by freeze / thaw method. MRNA was isolated by manual method. cDNA was obtained for apoptotic, autophagic and necrotic genes and the target gene DNA was amplified by real time-PCR method. These selected genes' expression were evaluated and determined important changes of depends on time in NaF induced molecular pathways. Apoptosis was induced by using apoptotic pathway of cells at 3 h, complete death of autophagic pathway at At the 3rd hour, cells were seen to enter apoptosis using apoptosis. The 12th hour deaths were completely autophagic. At the 24th hour, cell death was found to be complete by means of genes in the necrotic signal and apoptosis pathways in non-autophagous cells. As a result, it was determined that the molecular mechanisms of the NRF-52 renal cell lineage showed activity in NaF dependent cell deaths. It was concluded that the mechanisms of cytotoxic action were activated from the 3rd hour after the addition of NaF and different mechanisms accelerated cell death at 12th and 24th hours. Key words: NaF, kidney, cytotoxicity, cell culture