Hsa_circ_0007334通过海绵miR-144-3p促进人骨髓间充质干细胞成骨分化和增殖

IF 1.5 4区 医学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Critical Reviews in Eukaryotic Gene Expression Pub Date : 2023-01-01 DOI:10.1615/CritRevEukaryotGeneExpr.2022044929
Meng-Jun Liu, Bin Du, Jin-Song Yu, Ji Zhao, Hao Chen, Xing-Sheng Xiang, Yu-Zhu Wang, Wei Chen
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引用次数: 0

摘要

本研究旨在确定hsa_circ_0007334在人骨髓间充质干细胞(hBMSCs)成骨分化中的可能功能及其分子机制。采用实时荧光定量pcr (RT-qPCR)检测hsa_circ_0007334的水平。检测碱性磷酸酶(ALP)、RUNX2、成骨酶(OSX)、骨钙素(OCN),分析常规培养和hsa_circ_0007334控制下成骨分化程度。采用细胞计数试剂盒-8 (CCK-8)检测hBMSCs的增殖。采用Transwell法检测hBMSCs的迁移。利用生物信息学分析预测hsa_circ_0007334或miR-144-3p的可能靶标。采用双荧光素酶报告系统分析hsa_circ_0007334与miR-144-3p的结合。Hsa_circ_0007334在hBMSCs成骨分化中表达上调。通过ALP和骨标志物(RUNX2, OCN, OSX)水平证实hsa_circ_0007334体外成骨分化增强。hsa_circ_0007334过表达促进hBMSCs成骨分化、增殖和迁移,而敲低hsa_circ_0007334则具有相反的作用。miR-144-3p被确定为hsa_circ_0007334的靶标。miR-144-3p的靶基因参与成骨分化相关的生物学过程(如骨发育、上皮细胞增殖、间充质细胞凋亡过程)和通路(包括FoxO和VEGF信号通路)。因此,Hsa_circ_0007334是一种很有前途的成骨分化生物。
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Hsa_circ_0007334 Promotes the Osteogenic Differentiation and Proliferation of Human Bone Marrow Mesenchymal Stem Cells by Sponging miR-144-3p.

This study aimed to identify the possible function and the molecular mechanism of hsa_circ_0007334 in human bone marrow mesenchymal stem cells (hBMSCs) osteogenic differentiation. The level of hsa_circ_0007334 was detected by means of quantitative real-time polymerase chain reaction (RT-qPCR). Alkaline phosphatase (ALP), RUNX2, osterix (OSX), and osteocalcin (OCN) were monitored to analyze the degree of osteogenic differentiation under routine culture or under the control of hsa_circ_0007334. The proliferation of hBMSCs was tested with a cell counting kit-8 (CCK-8) assay. The migration of hBMSCs was tested using the Transwell assay. Bioinformatics analysis was used to predict the possible targets of hsa_circ_0007334 or miR-144-3p. Dual-luciferase reporter assay system was used to analyze the combination between hsa_circ_0007334 and miR-144-3p. Hsa_circ_0007334 was upregulated in osteogenic differentiation of hBMSCs. Osteogenic differentiation increased by hsa_circ_0007334 in vitro was confirmed with levels of ALP and bone markers (RUNX2, OCN, OSX). hsa_circ_0007334 overexpression promoted osteogenic differentiation, proliferation, and migration of hBMSCs, and knockdown of hsa_circ_0007334 has the opposite effects. miR-144-3p was identified as the target of hsa_circ_0007334. The targeting genes of miR-144-3p are involved in osteogenic-differentia-tion-related biological processes (such as bone development, epithelial cell proliferation, and mesenchymal cell apoptotic prosess) and pathways (including FoxO and VEGF signaling pathway). Hsa_circ_0007334, therefore, presents itself as a promising biological for osteogenic differentiation.

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来源期刊
Critical Reviews in Eukaryotic Gene Expression
Critical Reviews in Eukaryotic Gene Expression 生物-生物工程与应用微生物
CiteScore
2.70
自引率
0.00%
发文量
67
审稿时长
1 months
期刊介绍: Critical ReviewsTM in Eukaryotic Gene Expression presents timely concepts and experimental approaches that are contributing to rapid advances in our mechanistic understanding of gene regulation, organization, and structure within the contexts of biological control and the diagnosis/treatment of disease. The journal provides in-depth critical reviews, on well-defined topics of immediate interest, written by recognized specialists in the field. Extensive literature citations provide a comprehensive information resource. Reviews are developed from an historical perspective and suggest directions that can be anticipated. Strengths as well as limitations of methodologies and experimental strategies are considered.
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