Fluoride (F) is capable of promoting abnormal proliferation and differentiation in primary cultured mouse osteoblasts(OB cells), although; the underlying mechanism responsible remain rare. This study aimed to explore the roles… Click to show full abstract
Fluoride (F) is capable of promoting abnormal proliferation and differentiation in primary cultured mouse osteoblasts(OB cells), although; the underlying mechanism responsible remain rare. This study aimed to explore the roles of Wingless and INT-1(Wnt) signaling pathways and screen appropriate doses of calcium (Ca2+) to alleviate the sodium fluoride (NaF)-induced OB cells toxicity. For this, we evaluated the effect of Dickkopf-related protein 1 (DKK1) and Ca2+ on mRNA levels of wingless/integrated 3a (Wnt3a), low-density lipoprotein receptor-related protein 5 (LRP5), Dishevelled 1 (Dv1), glycogen synthase kinase 3β (GSK3β), β-catenin, lymphoid enhancer binding factor 1 (LEF1), and cellular myelocytomatosis oncogene (cMYC), as well as Ccnd1 (Cyclin D1) in OB cells challenged with 10-6 moL/L NaF for 24 h. Data demonstrated showed that F significantly increased the OB cells proliferation rate. Ectogenic 0.5 mg/L DKK1 significantly inhibited the proliferation of OB cells induced by F. The mRNA expression levels of Wnt 3a, LRP5, Dv1, LEF1, β-catenin, cMYC and Ccnd1 were significantly increased in F group, while significantly decreased in 10-6 moL/L NaF+0.5 mg/L DKK1 (FY) group. The mRNA expression levels of Wnt3a, LRP5, β-catenin and cMYC were significantly decreased in 10-6 moL/L NaF+2 mmoL/L CaCl2 (F+CaII) group. The proteins expression levels of Wnt3a, Cyclin D1, cMYC and β-catenin were significantly increased in F group whereas decreased in the F+CaII group. However, the mRNA and protein expression levels of GSK3β were significantly decreased in the F group while significantly increased in the F+CaII group. In summary, F activated the canonical Wnt/β-catenin pathway, changed the related genes expression and β-catenin protein location in OB cells, promoting cell proliferation. 2 mmoL/L Ca2+ supplementation reversed the expression levels of genes and proteins related to the canonical Wnt/β-catenin pathway.
               
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