piR-368 promotes odontoblastic differentiation of dental papilla cells via the Smad1/5 signaling pathway by targeting Smurf1.

IF 2.8 4区医学 Q3 CELL BIOLOGY

Connective Tissue Research Pub Date : 2024-01-01 Epub Date: 2024-02-01 DOI:10.1080/03008207.2023.2281319

Xinhui Xing, Yawei She, Guohua Yuan, Guobin Yang

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引用次数: 0

Abstract

Purpose: The important role of non-coding RNAs in odontoblastic differentiation of dental tissue-derived stem cells has been widely demonstrated; however, whether piRNA (a subclass of non-coding RNA) involved in the course of odontoblastic differentiation is not yet available. This study aimed to investigate the expression profile of piRNA during odontogenic differentiation of mDPCs and the potential molecular mechanism in vitro.

Materials and methods: The primary mouse dental papilla cells (mDPCs) were isolated from the first molars of 1-day postnatal Kunming mice. Then, they were cultured in odontogenic medium for 9 days. The expression profile of piRNA was detected by Small RNA sequencing. RT-qPCR was used to verify the elevation of piR-368. The mRNA and protein levels of mineralization markers were examined by qRT-PCR and Western blot analysis. Alkaline phosphatase (ALP) activity and alizarin red S staining were conducted to assess the odontoblastic differentiation ability.

Results: We validated piR-368 was significantly upregulated and interference with piR-368 markedly inhibited the odontogenic differentiation of mDPCs. In addition, the relationship between Smad1/5 signaling pathway and piR-368-induced odontoblastic differentiation has been discovered. Finally, we demonstrated Smurf1 as a target gene of piR-368 using dual-luciferase assays.

Conclusion: This study was the first to illustrate the participation of piRNA in odontoblastic differentiation. We proved that piR-368 promoted odontoblastic differentiation of mouse dental papilla cells via the Smad1/5 signaling pathway by targeting Smurf1.

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piR-368通过Smad1/5信号通路靶向Smurf1，促进牙乳头细胞成牙细胞分化。

目的:非编码rna在牙组织源性干细胞成牙细胞分化中的重要作用已被广泛证实;然而，piRNA(非编码RNA的一个亚类)是否参与成牙细胞分化过程尚不清楚。本研究旨在探讨piRNA在mDPCs成牙分化过程中的表达谱及其可能的体外分子机制。材料与方法:从昆明小鼠出生1 d后的第一磨牙中分离小鼠原代牙乳头细胞(mDPCs)。然后在成牙培养基中培养9 d。通过小RNA测序检测piRNA的表达谱。RT-qPCR检测mir -368的升高。采用qRT-PCR和Western blot检测矿化标志物mRNA和蛋白水平。以碱性磷酸酶(ALP)活性和茜素红S染色评价成牙细胞分化能力。结果:我们证实了piR-368显著上调，干扰piR-368可显著抑制mDPCs的成牙分化。此外，还发现了Smad1/5信号通路与pir -368诱导的成牙细胞分化之间的关系。最后，我们通过双荧光素酶测定证实了Smurf1是piR-368的靶基因。结论:本研究首次阐明了piRNA在成牙细胞分化中的作用。我们通过靶向Smurf1，证明了piR-368通过Smad1/5信号通路促进小鼠牙乳头细胞成牙细胞分化。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Connective Tissue Research 生物-细胞生物学

CiteScore

6.60

自引率

3.40%

发文量

审稿时长

2 months

期刊介绍： The aim of Connective Tissue Research is to present original and significant research in all basic areas of connective tissue and matrix biology. The journal also provides topical reviews and, on occasion, the proceedings of conferences in areas of special interest at which original work is presented. The journal supports an interdisciplinary approach; we present a variety of perspectives from different disciplines, including Biochemistry Cell and Molecular Biology Immunology Structural Biology Biophysics Biomechanics Regenerative Medicine The interests of the Editorial Board are to understand, mechanistically, the structure-function relationships in connective tissue extracellular matrix, and its associated cells, through interpretation of sophisticated experimentation using state-of-the-art technologies that include molecular genetics, imaging, immunology, biomechanics and tissue engineering.