LncRNA XIST facilitates the odontogenic differentiation of dental pulp stem cells via the FUS/ZBTB16.

IF 2.6 3区医学 Q2 DENTISTRY, ORAL SURGERY & MEDICINE

Journal of Applied Oral Science Pub Date : 2024-07-08 eCollection Date: 2024-01-01 DOI:10.1590/1678-7757-2023-0444

Ruiqing Cheng, Honglei Sun, Xiaotong Qiao, Xuefang Chen

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引用次数: 0

Abstract

Objective: This study aims to explore the regulatory mechanism of long noncoding RNA X inactive specific transcript (lncRNA XIST) in the odontogenic differentiation of human dental pulp stem cells (hDPSCs). hDPSCs were obtained from freshly extracted third molars and identified by flow cytometry.

Methodology: Odontogenic differentiation was induced in mineralized culture medium, and hDPSCs were infected with shRNA lentivirus targeting XIST or fused in sarcoma (FUS), followed by detection of alkaline phoshpatase (ALP) activity, alizarin red staining of mineralized nodules, Real-Time Quantitative Reverse Transcription PCR (qRT-PCR) quantification of XIST expression, and Western blot analysis of FUS, ZBTB16, and odontogenic differentiation markers (DSPP and DMP1). IF-FISH was performed to detect the cellular localization of XIST and FUS. RIP assay validated the XIST and FUS binding. ZBTB16 mRNA stability was tested after actinomycin D treatment. hDPSCs were infected with oe-ZBTB16 lentivirus and further treated with sh-XIST for a combined experiment.

Results: LncRNA XIST was highly expressed in hDPSCs with odontogenic differentiation. Downregulation of XIST or FUS weakened the ALP activity of hDPSCs, reduced mineralized nodules, diminished DSPP and DMP1 expressions. XIST binds to FUS to stabilize ZBTB16 mRNA and promote ZBTB16 expression. ZBTB16 overexpression partially reversed the inhibitory effect of XIST silencing on odontogenic differentiation of hDPSCs.

Conclusion: In conclusion, XIST stabilizes ZBTB16 mRNA and promotes ZBTB16 expression by binding to FUS, thereby facilitating the odontogenic differentiation of hDPSCs.

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LncRNA XIST通过FUS/ZBTB16促进牙髓干细胞的成牙性分化。

目的：本研究旨在探讨长链非编码RNA X失活特异性转录物（lncRNA XIST）在人牙髓干细胞（hDPSCs）成牙分化中的调控机制。从新鲜提取的第三磨牙中获得hdpsc，并通过流式细胞术进行鉴定。方法：在矿化培养基中诱导成牙分化，用靶向XIST或融合肉瘤（FUS）的shRNA慢病毒感染hdpsc，检测碱性磷酸酶（ALP）活性，对矿化结节进行茜素红染色，实时定量反转录PCR （qRT-PCR）定量检测XIST表达，Western blot分析FUS、ZBTB16和成牙分化标志物（DSPP和DMP1）。IF-FISH检测XIST和FUS的细胞定位。RIP实验证实了XIST和FUS的结合。放线菌素D处理后检测ZBTB16 mRNA的稳定性。用e- zbtb16慢病毒感染hDPSCs，再用sh-XIST进行联合实验。结果：LncRNA XIST在牙源性分化的hdpsc中高表达。XIST或FUS的下调使hdpsc的ALP活性减弱，矿化结节减少，DSPP和DMP1表达降低。XIST结合FUS稳定ZBTB16 mRNA，促进ZBTB16表达。ZBTB16过表达部分逆转了XIST沉默对hdpsc成牙分化的抑制作用。结论：综上所述，XIST通过与FUS结合稳定ZBTB16 mRNA，促进ZBTB16的表达，从而促进hdpsc的成牙性分化。

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

Journal of Applied Oral Science 医学-牙科与口腔外科

CiteScore

4.80

自引率

3.70%

发文量

审稿时长

4-8 weeks

期刊介绍： The Journal of Applied Oral Science is committed in publishing the scientific and technologic advances achieved by the dental community, according to the quality indicators and peer reviewed material, with the objective of assuring its acceptability at the local, regional, national and international levels. The primary goal of The Journal of Applied Oral Science is to publish the outcomes of original investigations as well as invited case reports and invited reviews in the field of Dentistry and related areas.