MicroRNA-204 regulates osteogenic induction in dental follicle cells.
MicroRNA-204 regulates osteogenic induction in dental follicle cells.
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DOI：
10.1016/j.jds.2019.11.004
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发表时间：
2020-12
影响因子：
3.5
通讯作者：
Kondoh T
中科院分区：
文献类型：
--
作者：
Ito K;Tomoki R;Ogura N;Takahashi K;Eda T;Yamazaki F;Kato Y;Goss A;Kondoh T
牙囊是一种外胚间充质组织，包围着发育中的牙胚，含有成骨细胞系定向的干/祖细胞。MicroRNAs(MiRNAs)是一种小的非编码RNA，在干细胞生长、增殖和分化过程中调节基因表达。本研究旨在探讨miRNA在人牙囊细胞成骨分化过程中的关键调控作用。我们分析了成骨细胞分化过程中hDFC中miRNA的表达谱。基因芯片分析显示，在成骨诱导过程中，hDFC中miR-204的表达降低。实时荧光定量和RT-PCR分析还显示，在成骨分化过程中，miR-204在三种HDFC中的表达均降低。为了研究miR-204是否对成骨分化有影响，通过电子分析预测miR-204靶向碱性磷酸酶(ALP)、富含半胱氨酸的酸性分泌蛋白(SPARC)和3‘-UTRs中的Runx2。将miR-204基因导入HDFC后，细胞内ALP活性降低，SPARC和Runx2蛋白表达水平降低。MiR-204基因表达下调了ALP、SPARC和Runx2基因的表达水平。我们的结果提示miR-204通过靶向骨特异性转录因子Runx2、矿化标志物ALP和骨细胞外基质蛋白SPARC来负向调节HDFC的成骨分化。
The dental follicle is an ectomesenchymal tissue surrounding developing tooth germ that contains osteoblastic-lineage-committed stem/progenitor cells. MicroRNAs (miRNAs) are small non-coding RNAs that regulate gene expression during stem cell growth, proliferation, and differentiation. The aim of this study was to investigate the key regulators of miRNA during osteogenic differentiation in human dental follicle cells (hDFC). We analyzed miRNA expression profiles in hDFC during osteoblastic differentiation. Expression of miR-204 was decreased in hDFC during osteogenic induction on microarray analysis. Real-time and RT-PCR analysis also showed that the expression of miR-204 was decreased in all three hDFC during osteogenic differentiation. To investigate whether miR-204 has an effect on osteogenic differentiation, miR-204 was predicted to target alkaline phosphatase (ALP), secreted protein acidic and rich in cysteine (SPARC), and Runx2 in the in the 3'-UTRs by in silico analysis. When miR-204 was transfected into hDFC, the activity of ALP and protein levels of SPARC and Runx2 were decreased. mRNA levels of ALP, SPARC and Runx2 were also decreased by miR-204 transfection. Our data suggest that miR-204 negatively regulates the osteogenic differentiation of hDFC by targeting the bone-specific transcription factor Runx2, the mineralization maker ALP and the bone extracellular matrix protein SPARC.
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