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The Effects of Sodium Fluoride (NaF) Treatment on the PI3K/Akt Signal Pathway in NRK-52E Cells

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Abstract

The effects of the element fluorine on the phosphoinositide-3-kinase–protein kinase B/Akt (PI3K/Akt) pathway has a significant role in regulation of intracellular molecular mechanisms. NRK-52E rat kidney epithelial cell line was selected as the material of the study. NaF was used as the fluorine source in the study. The NaF dose was determined with the MTT assay. The NaF concentrations were determined as the proliferation concentration of 10 μM and IC25 (2250 μM) and IC50 (4250 μM) for 24 h. In the study, the erb-b2 receptor tyrosine kinase 2 (ERBB2), phosphoinositide-3-kinase (PI3K), Protein kinase B (PKB,Akt), Mammalian target of rapamycin (mTOR), and the Tumor protein 53 (TP53) genes were considered as the target genes. NaF concentration was administered on the cells. Total mRNA was isolated. mRNAs were turned into cDNA. The expression levels of the target genes were determined by RT-qPCR method. According to the results obtained in the study, the low NaF concentration increased the expression levels of the ERBB2, PI3K, and Akt genes, while the higher concentrations did not significantly affect these levels. The expression of mTOR decreased at all given concentrations. The expression of the TP53 gene did not change at the low concentration, while it increased at the high concentrations. Based on the results, it may be stated that fluorine may inhibit the kinase enzymes in the PI3K/Akt pathway. In summary, in the pathogenesis of the cell damage caused by fluorine in the NRK-52E cell line, the PI3K/Akt/mTOR pathway is an important signal pathway.

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Data Availability

The datasets used or analyzed in this study are available from the corresponding author on reasonable request.

References

  1. Song GH, Gao JP, Wang CF, Chen CY, Yan XY, Guo M, Wang Y, Huang FB (2014) Sodium fluoride induces apoptosis in the kidney of rats through caspase-mediated pathways and DNA damage. J Physiol Biochem 70(3):857–868

    Article  CAS  Google Scholar 

  2. Ergin E, Eden E (2017) Does fluorine have a negative effect on human health? J Ege Uni Faculty of Dent 201738(1):13–20

    Google Scholar 

  3. Agalakova NI, Gusev GP (2012) Molecular mechanisms of cytotoxicity and apoptosis induced by inorganic fluoride. Int Schol Res 2012:Article ID 403835

  4. Song GH, Huang FB, Gao JP, Liu ML, Pang WB, Wb Li, Yan XY, Huo MJ, Yang X (2015) Effects of fluoride on DNA damage and caspase-mediated apoptosis in the liver of rats. Biol Trace Elem Res 166(2):173–182

    Article  CAS  Google Scholar 

  5. Caglayan C, Kandemir FM, Darendelioğlu E, Küçükler S, Ayna A (2021) Hesperidin protects liver and kidney against sodium fluoride-induced toxicity through anti-apoptotic and anti-autophagic mechanisms. Life Sci 281:119730

    Article  CAS  Google Scholar 

  6. Hemmings BA, Restuccia DF (2012) Pi3k-pkb/akt pathway. Cold Spring Harbor perspectives in biology 4(9):a011189

    Article  Google Scholar 

  7. Yüksek V, Dede S, Taşpınar M, Çetin S (2017) The effects of certain vitamins on apoptosis and DNA damage in sodium fluoride (NaF) administered renal and osteoblast cell lines. Fluoride 50(3):300–313

    Google Scholar 

  8. Chomczynski P, Mackey K (1995) Modification of the TRI reagent procedure for isolation of RNA from polysaccharide- and proteoglycan-rich sources. Biotechniques 19(6):942–945

    CAS  PubMed  Google Scholar 

  9. Livak KJ, Schmittgen TD (2011) Analysis of relative gene expression data using real-time quantitative PCR and the 2− ΔΔCT method. Methods 25(4):402–408

    Article  Google Scholar 

  10. Anuradha CD, Kanno S, Hirano S (2001) Oxidative damage to mitochondria is a preliminary step to caspase-3 activation in fluoride-induced apoptosis in HL-60 cells. Free Radical Biol Med 1:367–373

    Article  Google Scholar 

  11. Bai C, Chen T, Cui Y, Gong T, Peng X, Cui HM (2010) Effect of high fluorine on the cell cycle and apoptosis of renal cells in chickens. Biol Trace Elem Res 138(1–3):173–180

    Article  CAS  Google Scholar 

  12. Barbier O, Arreolar-Mendoza L, Del Roza LM (2010) Molecular mechanisms of fluoride toxicity. Chem-Biological Interactions 188:319–333

    Article  CAS  Google Scholar 

  13. Xu H, JinXQ JL, Li GS (2006) Effect of sodium fluoride on the expression of bcl2 family and osteopontin in rat renal tubular cells. Biol Trace Elem Res 109(1):55–60

    Article  CAS  Google Scholar 

  14. Guney M, Oral B, Demirin H, Karahan N, Mungan T, Delibas N (2007) Protective effects of vitamins C and E against endometrial damage and oxidative stress in fluoride intoxication. Clin Exp Pharmacol Physiol 34(5–6):467–674

    Article  CAS  Google Scholar 

  15. Çetin S, Yur F, Taşpinar M, Yüksek V (2019) The effects of some minerals on apoptosis and DNA damage in sodium fluoride-administered renal and osteoblast cell lines. Fluoride 52(3):362–378

    Google Scholar 

  16. Yüksek V, Dede S, Usta A, Çetın S, Taşpınar M (2020) DNA damage-induced by sodium fluoride (NaF) and the effect of cholecalciferol. Biocell 44(2):263–268

    Article  Google Scholar 

  17. Pompura SL, Dominguez-Villar M (2018) The PI3K/AKT signaling pathway in regulatory t-cell development, stability, and function. J Leukoc Biol 103:1065–1076

    Article  CAS  Google Scholar 

  18. He H, Wang H, Jiao Y, Ma C, Zhang H, Zhou Z (2015) Effect of sodium fluoride on the proliferation 352 and gene differential expression in human RPMI8226 Cells. Biol Trace Elem Res 167(1):11–17

    Article  CAS  Google Scholar 

  19. Otsuki S, Morshed SR, Chowdhury SA, Takayama F, Satoh T, Hashimoto K, Sugiyama K, Amano O, Yasui T, Yokote Y, Akahane K, Sakagami H (2005) Possible link between glycolysis and apoptosis induced by sodium fluoride. J Dent Res 84(10):919–923

    Article  CAS  Google Scholar 

  20. Gutowska I, Baranowska-Bosiacka I, Siwiec E, Szczuko M, Kolasa A, Kondarewicz A et al (2016) Lead enhances fluoride influence on apoptotic processes in the HepG2 liver cell line. Toxicol Ind Health 32(3):517–525

    Article  CAS  Google Scholar 

  21. Viñals F, Testar X, Palacín M, Zorzano A (1993) Inhibitory effect of fluoride on ınsulin receptor autophosphorylation and tyrosine kinase activity. Biochem J 291(Pt 2):615–622

    Article  Google Scholar 

  22. Urut F, Dede S, Yuksek V, Cetin S, Usta A, Taspinar M (2021) In vitro evaluation of the apoptotic, autophagic, and necrotic molecular pathways of fluoride. Biol Trace Elem Res 199(10):3700–3706

    Article  CAS  Google Scholar 

  23. Efe U, Dede S, Yüksek V, Çetin S (2021) Apoptotic and oxidative mechanisms in liver and kidney tissues of sheep with fluorosis. Biol Trace Elem Res 199:136–141

    Article  CAS  Google Scholar 

  24. Ameeramja J, Panneerselvam L, Govindarajan V, Jeyachandran S, Baskaralingam V, Perumal E (2016) Tamarind seed coat ameliorates fluoride induced cytotoxicity, oxidative stress, mitochondrial dysfunction and apoptosis in A549 cells. J Hazard Mater 15(301):554–565

    Article  Google Scholar 

  25. Penuel E, Schaefer G, Akita RW, Sliwkowski MX (2001) Structural requirements for ErbB2 transactivation. Semin Oncol 28(6 Suppl 18):36–42

    Article  CAS  Google Scholar 

  26. Wichmann H, Güttler A, Bache M, Taubert H, Vetter M, Würl P, Holzhausen HJ, Eckert AW, Kappler M, Vordermark D (2014) Inverse prognostic ımpact of ErbB2 mRNA and protein expression level in tumors of soft tissue sarcoma patients. Strahlenther Onkol 190(10):912–918

    Article  Google Scholar 

  27. Zhou BH, Tan PP, Jia LS, Zhao WP, Wang JC, Wang HW (2018) PI3K/AKT signaling pathway involvement in fluoride-induced apoptosis in C2C12 cells. Chemosphere 199:297–302

    Article  CAS  Google Scholar 

  28. Kuang P, Deng H, Liu H, Cui H, Fang J, Zuo Z, Deng J, Li Y, Wang X, Zhao L (2018) Sodium fluoride induces splenocyte autophagy via the mammalian targets of rapamycin (mTOR) signaling pathway in growing mice. Aging (Albany NY) 10(7):1649–1665

    Article  CAS  Google Scholar 

  29. Huang Y, Sun M, Li F, Li H, Jiang Z (2018) Preliminary study of mechanisms of fluoride-induced suppression of nitric oxide synthesis in human umbilical vein endothelial cells. Biol Trace Elem Res 185(2):311–315

    Article  CAS  Google Scholar 

  30. Fan B, Yu Y, Zhang Y (2015) PI3K-Akt1 expression and its significance in liver tissues with chronic fluorosis. Int J Clin Exp Pathol 8(2):1226–1236

    PubMed  PubMed Central  Google Scholar 

  31. Shenoy PS, Sen U, Kapoor S, Ranade AV, Chowdhury CR, Bose B (2019) Sodium fluoride induced skeletal muscle changes: degradation of proteins and signaling mechanism. Environ Pollut 244:534–548

    Article  CAS  Google Scholar 

  32. Guo Q, Sun Z, Niu R, ManthariRK Yuan M, Yang K, Cheng M, Gong Z, Wang J (2020) Effect of arsenic and/or fluoride gestational exposure on renal autophagy in offspring mice. Chemosphere 241:124861

    Article  CAS  Google Scholar 

  33. Zhang J, Zhu Y, Shi Y, Han Y, Liang C, Feng Z, Zheng H, Eng M, Wang J (2017) Fluoride-induced autophagy via the regulation of phosphorylation of mammalian targets of rapamycin in mice leydig cells. J Agric Food Chem 65(40):8966–8976

    Article  CAS  Google Scholar 

  34. Lei S, Zhang Y, Zhang K, Li J, Liu L (2015) Effects of fluoride on the expression of Beclin1 and mTOR in ameloblasts. Cells Tissues Organs 200(6):405–412

    Article  Google Scholar 

  35. Suzuki M, Ikeda A, Bartlett JD (2018) Sirt1 overexpression suppresses fluoride-induced p53 acetylation to alleviate fluoride toxicity in ameloblasts responsible for enamel formation. Arch Toxicol 92(3):1283–1293

    Article  CAS  Google Scholar 

  36. Tu W, Zhang Q, Liu Y, Han L, Wang Q, Chen P, Zhang S, Wang A, Zhou X (2018) Fluoride induces apoptosis via inhibiting SIRT1 activity to activate mitochondrial p53 pathway in human neuroblastoma SH-SY5Y cells. Toxicol Appl Pharmacol 347:60–69

    Article  CAS  Google Scholar 

  37. Luo Q, Guo H, Kuang P, Cui H, Deng H, Liu H, Lu Y, Wei Q, Chen L, Fang J, Zuo Z, Deng J, Li Y, Wang X, Zhao L (2018) Sodium fluoride arrests renal G2/M phase cell-cycle progression by activating ATM-Chk2-P53/Cdc25C signaling pathway in mice. Cell Physiol Biochem 51(5):2421–2433

    Article  CAS  Google Scholar 

  38. Wen P, Wei X, Liang G, Wang Y, Yang Y, Qin L, Pang W, Qin G, Li H, Jiang Y, Wu Q (2019) Long-term exposure to low level of fluoride induces apoptosis via p53 pathway in lymphocytes of aluminum smelter workers. Environ Sci Pollut Res Int 26(3):2671–2680

    Article  CAS  Google Scholar 

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Funding

This study was supported by a Grant from the Scientific Research Projects Presidency of Van Yüzüncü Yil University (TYL-2019–822.

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All authors contributed to the study’s conception and design. Material preparation, data collection, and analysis were performed by Rıskiye Korkmaz, Veysel Yüksek, and Semiha Dede. The first draft of the manuscript was written by Rıskiye Korkmaz, and all authors commented on the previous versions of the manuscript. All authors read and approved the final manuscript.

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Correspondence to Veysel Yüksek.

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Korkmaz, R., Yüksek, V. & Dede, S. The Effects of Sodium Fluoride (NaF) Treatment on the PI3K/Akt Signal Pathway in NRK-52E Cells. Biol Trace Elem Res 200, 3294–3302 (2022). https://doi.org/10.1007/s12011-021-02927-4

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