Invitro investigation of theeffect of sodium fluoride (NAF) administration on PI3K/AKT signaling pathway

Korkmaz R, Invitro investigation of theeffect of sodium fluoride (NaF) administration on PI3K/AKT signaling pathway. Health Sciences Institute, Department of Biochemistry (Veterinary Medicine), M.Sc. Thesis, Van, 2020. This thesis study was planned to determine the effect of fluoride has high electronegativity on the PI3K/AKT pathway, which has an important role in the regulation of intracellula rmolecular mechanisms. For this purpose, NRK-52E rat kidney epithelial cell was chosen as the study material, since kidney tissue is the leading tissue the most affected by fluoride. NaF was used as a source of fluorine in thestudy. The NaF concentration to be applied was determined byt he MTT viability test. NaF concentrations were determined as concentrations (10 μM) which are proliferation enhancing, IC25 (2250 μM) and IC50 (4250 μM) cytotoxic concentrations for 24 hours, and were applied. In the study, genes ERBB2, PI3K, AKT and mTOR genes , an important role in PI3K/ AKT pathways , and TP53 genes that have pathway were considered as target genes. The determined NaF concentration was applied to the cells and at the end of 24 hours. Total mRNA was isolated in the study groups. The obtained mRNAs were translated into cDNA. Expression levels of target genes were determined by real time-qPCR method by using the cDNAs. According to the results obtained at the end of the study, it was determined that the low concentration of NaF increased the expression of ERBB2, PI3K, AKT genes, but did not change in high concentrations. It was found that mTOR decreased in all NaF concentrations given. TP53 gene was found to be unchanged at low concentration, but increased in highs concentration. According to the results obtained from the study and the literature, it was concluded that fluoride can inhibit the kinase enzymes in the PI3K/AKT pathway and as a result, it may cause a decrease or increase in the expression of somegenes. In summary, the PI3K/AKT/mTOR is an important signaling pathway in the pathogenesis of fluorine-induced cell damage in the NRK-52E renal epithelial cell line.