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Preliminary Analysis of MicroRNAs Expression Profiling in MC3T3-E1 Cells Exposed to Fluoride

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Abstract

Overexposure to fluoride from environmental sources can cause serious public health problems. Disrupted osteoblast function and impaired bone formation were found to be associated with excessive fluoride exposure. A massive analysis of microRNAs (miRNAs) was used to figure out the possible pathways in which fluoride affects osteoblast function. MC3T3-E1 cells were treated with 8 mg/L of fluorine for 7 days. Total RNA of cells was extracted, and their integrity and purity were tested. RNA samples were analyzed by using miRNA array, including miRNA labeling, hybridization, scanning, and expression data analysis to compare the profiling of miRNA expression between control and fluoride-treated group. Transcriptome analysis console and enrichment analysis calculated by miRSystem were used to predict target genes and collect miRNAs pathway maps. Forty-five upregulated and 31 downregulated miRNAs expression were found in the fluoride-treated group, and most of the verified miRNAs were mature. The KEGG pathway enrichment analysis searched out 36 pathways that scored more than 0.1. These pathways mainly included intracellular signaling, cytokines, metabolism, and cytoskeleton-related pathways. Among them, the Wnt, insulin, TGF-beta, hedgehog, VEGF, and notch pathways in osteoblasts were those mainly affected by fluoride treatment. These results have shown a number of higher level systemic pathways activated by overexposure of fluoride in osteoblastic cells and verified that fluoride affected the molecular crosstalk in the osteoblasts.

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References

  1. Sarkis KS, PinheiroMde M, Szejnfeld VL, et al. (2012) High bone density and bone health. Endocrinol Nutr 59(3):207–214

    Article  PubMed  Google Scholar 

  2. Kanis JA (1993) Treatment of symptomatic osteoporosis with fluoride. Am J Med 95(5A):53S–61S

    Article  CAS  PubMed  Google Scholar 

  3. Kurdi MS (2016) Chronic fluorosis: the disease and its anaesthetic implications. Indian J Anaesth 60(3):157–162

    Article  PubMed  PubMed Central  Google Scholar 

  4. Gupta SK, Gambhir S, Mithal A, et al. (1993) Skeletal scintigraphic findings in endemic skeletal fluorosis.Nucl. Med Commun 14(5):384–390

    Article  CAS  Google Scholar 

  5. Sun F, Li X, Yang C, et al. (2014) Role for PERK in the mechanism underlying fluoride-induced bone turnover. Toxicology 325:52–66

    Article  CAS  PubMed  Google Scholar 

  6. Li XN, Lv P, Sun Z, et al. (2014) Role of unfolded protein response in affecting osteoblast differentiation induced by fluoride. Biol Trace Elem Res 158(1):113–121

    Article  CAS  PubMed  Google Scholar 

  7. Chen CZ, Li L, Lodish HF, et al. (2004) MicroRNAs modulate hematopoietic lineage differentiation. Science 303(5654):83–86

    Article  CAS  PubMed  Google Scholar 

  8. Tuddenham L, Wheeler G, Ntounia-Fousara S, et al. (2006) The cartilage specific microRNA-140 targets histone deacetylase 4 in mouse cells. FEBS Lett 580(17):4214–4217

    Article  CAS  PubMed  Google Scholar 

  9. Sun M, Zhou X, Chen L, et al. (2016) The regulatory roles of microRNAs in bone remodeling and perspectives as biomarkers in osteoporosis. Biomed Res Int 2016:1652417

    PubMed  PubMed Central  Google Scholar 

  10. Fan J, Li J, Fan Q (2015) Naringin promotes differentiation of bone marrow stem cells into osteoblasts by upregulating the expression levels of microRNA-20a and downregulating the expression levels of PPARγ. Mol Med Rep 12(3):4759–4765

    CAS  PubMed  Google Scholar 

  11. Li Z, Hassan MQ, Jafferji M, et al. (2009) Biological functions of miR-29b contribute to positive regulation of osteoblast differentiation. J BiolChem 284(23):15676–15684

    CAS  Google Scholar 

  12. Eskildsen T, Taipaleenmäki H, Stenvang J, et al. (2011) MicroRNA-138 regulates osteogenic differentiation of human stromal (mesenchymal) stem cells in vivo. Proc Natl Acad Sci U S A 108(15):6139–6144

    Article  PubMed  PubMed Central  Google Scholar 

  13. TP L, Lee CY, Tsai MH, et al. (2012) miRSystem: an integrated system for characterizing enriched functions and pathways of microRNA targets. PLoS One 7(8):e42390

    Article  Google Scholar 

  14. 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

    CAS  PubMed  PubMed Central  Google Scholar 

  15. Sun Z, Zhang W, Li S, et al. (2016) Altered miRNAs expression profiling in sperm of mice induced by fluoride. Chemosphere 155:109–114

    Article  CAS  PubMed  Google Scholar 

  16. CY H, Ren LQ, Li XN, et al. (2012) Effect of fluoride on insulin level of rats and insulin receptor expression in the MC3T3-E1 cells. Biol Trace Elem Res 150(1–3):297–305

    Google Scholar 

  17. Deng C, Yu Y (2014) Roles of hedgehog signaling pathway on injury of bone with fluorosis. Zhonghua Bing Li XueZaZhi 43(1):68–70

    CAS  Google Scholar 

  18. Yorgan T, Vollersen N, Riedel C, et al. (2016) Osteoblast-specific Notch2 inactivation causes increased trabecular bone mass at specific sites of the appendicular skeleton. Bone 87:136–146

    Article  CAS  PubMed  Google Scholar 

  19. Dicarlo M, Bianchi N, Ferretti C, et al. (2016) Evidence supporting a paracrine effect of IGF-1/VEGF on human mesenchymal stromal cell commitment. Cells Tissues Organs 201(5):333–341

    Article  PubMed  Google Scholar 

Download references

Acknowledgments

This work was supported by grant for skeletal fluorosis research from National Natural Science Foundation of China [81673111], the Norman Bethune Program of Jilin University [2012222], and Specialized Research Fund for the Doctoral Program of Higher Education [20130061110084].

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Correspondence to Hui Xu.

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Wang, Y., Zhang, X., Zhao, Z. et al. Preliminary Analysis of MicroRNAs Expression Profiling in MC3T3-E1 Cells Exposed to Fluoride. Biol Trace Elem Res 176, 367–373 (2017). https://doi.org/10.1007/s12011-016-0833-x

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  • DOI: https://doi.org/10.1007/s12011-016-0833-x

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