MicroRNA-3064-3p Regulates the Differentiation of Cementoblasts Through Targeting DKK1
Journal of periodontal research(2018)
摘要
Background and ObjectiveMicroRNAs (miRNAs) are short, noncoding RNAs that interfere with translation of target mRNAs and thereby play a pivotal role in a variety of biological processes. Cementoblasts are the cells that build up cementum. They share a similar gene expression pattern with osteoblasts. Recent studies have suggested that miRNAs are able to control osteoblast-mediated bone formation. However, the effects of miRNA on cementoblast differentiation still remain unsolved. Herein, we wanted to elucidate the role of miR-3064-3p in cementoblast differentiation. Material and MethodsA miRNA microarray was operated to explore the miRNA expression patterns during cementoblast differentiation. miR-3064-3p agomir/antagomir was used to promote or inhibit, respectively, the expression of miR-3064-3p. In order to measure the differentiation level of cementoblasts, quantitative RT-PCR (qRT-PCR), Alizarin red staining, and assessment of alkaline phosphatase activity were performed. Luciferase assays, qRT-PCR, and western blotting were used to identify the target gene of miR-3064-3p. ResultsmiR-3064-3p showed persistently decreased expression during cementoblast differentiation. Overexpression of miR-3064-3p suppressed cementoblast differentiation, while inhibition of miR-3064-3p promoted cementoblast differentiation. Target prediction-analysis tools and dual-luciferase assay identified Dickkopf WNT signaling pathway inhibitor 1 (DKK1) as a direct target of miR-3064-3p. Results from qRT-PCR and western blotting showed that inhibition of miR-3064-3p led to a remarkable increase in DKK1/Dickkopf related protein 1 (Dkk-1) expression. In addition, pretreatment with recombinant Dickkopf related protein 1 (Dkk-1) rescued the miR-3064-3p-mediated suppression of cementoblast differentiation. ConclusionThis study demonstrates, for the first time, that miR-3064-3p suppresses cementoblast differentiation via the regulation of DKK1.
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关键词
DKK1,cementoblast,microRNA,differentiation
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