DLX6-AS1 regulates odonto/osteogenic differentiation in dental pulp cells under the control of BMP9 via the miR-128-3p/MAPK14 axis: A laboratory investigation

IF 5.4 1区医学 Q1 DENTISTRY, ORAL SURGERY & MEDICINE

International endodontic journal Pub Date : 2024-07-07 DOI:10.1111/iej.14120

Liu Liu, Tongfeng Fang, Cheng Miao, Xiangfen Li, Yanglin Zeng, Tianyi Wang, Yubin Cao, Dingming Huang, Dongzhe Song

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引用次数: 0

Abstract

Aim

The regenerative capacity of dental pulp relies on the odonto/osteogenic differentiation of dental pulp cells (DPCs), but dynamic microenvironmental changes hinder the process. Bone morphogenetic protein 9 (BMP9) promotes differentiation of DPCs towards an odonto/osteogenic lineage, forming dentinal-like tissue. However, the molecular mechanism underlying its action remains unclear. This study investigates the role of DLX6 antisense RNA 1 (DLX6-AS1) in odonto/osteogenic differentiation induced by BMP9.

Methodology

Custom RT² profiler PCR array, quantitative Real-Time PCR (qRT-PCR) and western blots were used to investigate the expression pattern of DLX6-AS1 and its potential signal axis. Osteogenic ability was evaluated using alkaline phosphatase and alizarin red S staining. Interactions between lncRNA and miRNA, as well as miRNA and mRNA, were predicted through bioinformatic assays, which were subsequently validated via RNA immunoprecipitation and dual luciferase reporter assays. Student's t-test or one-way ANOVA with post hoc Tukey HSD tests were employed for data analysis, with a p-value of less than .05 considered statistically significant.

Results

DLX6-AS1 was upregulated upon BMP9 overexpression in DPCs, thereby promoting odonto/osteogenic differentiation. Additionally, miR-128-3p participated in BMP9-induced odonto/osteogenic differentiation by interacting with the downstream signal MAPK14. Modifying the expression of miR-128-3p and transfecting pcMAPK14/siMAPK14 had a rescue impact on odonto/osteogenic differentiation downstream of DLX6-AS1. Lastly, miR-128-3p directly interacted with both MAPK14 and DLX6-AS1.

Conclusions

DLX6-AS1 could regulate the odonto/osteogenic differentiation of DPCs under the control of BMP9 through the miR-128-3p/MAPK14 axis.

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DLX6-AS1通过miR-128-3p/MAPK14轴调控牙髓细胞在BMP9控制下的牙髓/骨生成分化：实验室研究

目的：牙髓的再生能力依赖于牙髓细胞（DPCs）的牙髓/骨生成分化，但动态的微环境变化阻碍了这一过程。骨形态发生蛋白9（BMP9）可促进牙髓细胞向牙髓/骨生成系分化，形成牙本质样组织。然而，其作用的分子机制仍不清楚。本研究探讨了DLX6反义RNA 1（DLX6-AS1）在BMP9诱导的牙性/骨性分化中的作用：方法：使用定制的RT2 profiler PCR阵列、定量实时PCR（qRT-PCR）和Western印迹研究DLX6-AS1的表达模式及其潜在的信号轴。碱性磷酸酶和茜素红 S 染色法评估了成骨能力。通过生物信息分析预测了lncRNA与miRNA以及miRNA与mRNA之间的相互作用，随后通过RNA免疫沉淀和双荧光素酶报告实验进行了验证。数据分析采用学生 t 检验或单向方差分析，并进行事后 Tukey HSD 检验，P 值小于 0.05 为具有统计学意义：结果：当BMP9在DPCs中过表达时，DLX6-AS1被上调，从而促进畸形/骨化分化。此外，miR-128-3p 通过与下游信号 MAPK14 相互作用，参与了 BMP9 诱导的骨头/骨生成分化。改变 miR-128-3p 的表达和转染 pcMAPK14/siMAPK14 对 DLX6-AS1 下游的畸牙/骨生成分化有挽救作用。最后，miR-128-3p直接与MAPK14和DLX6-AS1相互作用：结论：DLX6-AS1可在BMP9的控制下通过miR-128-3p/MAPK14轴调控DPCs的骨性/骨生成分化。

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

International endodontic journal 医学-牙科与口腔外科

CiteScore

10.20

自引率

28.00%

发文量

195

审稿时长

4-8 weeks

期刊介绍： The International Endodontic Journal is published monthly and strives to publish original articles of the highest quality to disseminate scientific and clinical knowledge; all manuscripts are subjected to peer review. Original scientific articles are published in the areas of biomedical science, applied materials science, bioengineering, epidemiology and social science relevant to endodontic disease and its management, and to the restoration of root-treated teeth. In addition, review articles, reports of clinical cases, book reviews, summaries and abstracts of scientific meetings and news items are accepted. The International Endodontic Journal is essential reading for general dental practitioners, specialist endodontists, research, scientists and dental teachers.