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.



Similar content being viewed by others
References
Sarkis KS, PinheiroMde M, Szejnfeld VL, et al. (2012) High bone density and bone health. Endocrinol Nutr 59(3):207–214
Kanis JA (1993) Treatment of symptomatic osteoporosis with fluoride. Am J Med 95(5A):53S–61S
Kurdi MS (2016) Chronic fluorosis: the disease and its anaesthetic implications. Indian J Anaesth 60(3):157–162
Gupta SK, Gambhir S, Mithal A, et al. (1993) Skeletal scintigraphic findings in endemic skeletal fluorosis.Nucl. Med Commun 14(5):384–390
Sun F, Li X, Yang C, et al. (2014) Role for PERK in the mechanism underlying fluoride-induced bone turnover. Toxicology 325:52–66
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
Chen CZ, Li L, Lodish HF, et al. (2004) MicroRNAs modulate hematopoietic lineage differentiation. Science 303(5654):83–86
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
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
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
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
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
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
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
Sun Z, Zhang W, Li S, et al. (2016) Altered miRNAs expression profiling in sperm of mice induced by fluoride. Chemosphere 155:109–114
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
Deng C, Yu Y (2014) Roles of hedgehog signaling pathway on injury of bone with fluorosis. Zhonghua Bing Li XueZaZhi 43(1):68–70
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
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
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].
Author information
Authors and Affiliations
Corresponding author
Electronic Supplementary Material
ESM 1
(XLSX 14 kb)
Rights and permissions
About this article
Cite this article
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
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12011-016-0833-x


