LncRNA AC003090.1 介导 miR-203a-3p/FOXP1 轴促进人骨髓间充质干细胞的成骨分化

IF 2.9 4区 医学 Q1 Medicine Journal of biomedical nanotechnology Pub Date : 2024-02-01 DOI:10.1166/jbn.2024.3744
Huafeng Zhuang, Xuedong Yao, Aifei Wang, Junjie Li, Miao Zheng, Yu Dong, Youjia Xu, Yizhong Li, Yongjun Lin
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引用次数: 0

摘要

我们研究了lncRNA AC003090.1在人类骨髓干细胞(hBMSCs)中的调控机制。我们收集了骨质疏松症(OP)患者的组织,通过 QPCR 检测和分析了 AC003090.1、miR-203a-3p 和 FOXP1 的表达。使用改良的 Gomori 钙钴测定法和 PNP 比色法检测成骨标志物碱性磷酸酶(ALP)的表达和活性。茜素红染色法显示了细胞外表面的钙沉积。使用油红 O 染色法检测细胞内脂质含量。双荧光素酶报告系统验证了 AC003090.1 或 FOXP1 与 miR-203a-3p 之间的靶向关系。 qRT-PCR 和 Western 印迹用于测量不同干预处理后 hBMSCs 中 β-catenin 的表达水平。在骨质疏松症(OP)中,AC003090.1和FOXP1的表达水平下调,而miR-203a-3p的表达水平上调。AC003090.1表达的增加可增强hBMSC的成骨分化(OD),并降低hBMSC的成脂能力。此外,miR-203a-3p模拟物或FOXP1基因敲除可逆转AC003090.1表达增加对hBMSCs成骨分化和成脂分化的影响。双荧光素酶报告实验表明,AC003090.1可以海绵状表达miR-203a-3p,而miR-203a-3p靶向hBMSCs中的FOXP1。LncRNA AC003090.1通过调节miR-203a-3p/FOXP1轴促进hBMSCs的OD。
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LncRNA AC003090.1 Mediates miR-203a-3p/FOXP1 Axis to Promote the Osteogenic Differentiation of Human Bone Marrow Mesenchymal Stem Cells
We investigated the regulatory mechanism of lncRNA AC003090.1 in human bone marrow stem cells (hBMSCs). Tissues from patients with osteoporosis (OP) were collected for the detection and analysis of AC003090.1, miR-203a-3p, and FOXP1 expression by QPCR. The expression and activity of alkaline phosphatase (ALP), an osteogenic marker, were detected using a modified Gomori calcium-cobalt assay and PNP colorimetric assay. Calcium deposition on the extracellular surface was demonstrated using alizarin red staining. Using the Oil Red O staining assay to detecte the Intracellular lipid content. Dual luciferase reporting system verified the targeting relationship between AC003090.1 or FOXP1 and miR-203a-3p. qRT-PCR and Western blot were used to measure the expression level of β-catenin in hBMSCs after different intervention treatments. The expression levels of AC003090.1 and FOXP1 were downregulated in Osteoporosis (OP), whereas those of miR-203a-3p were upregulated. An increase in AC003090.1 expression could enhance hBMSC osteogenic differentiation (OD) and reduce the adipogenic ability of hBMSCs. Furthermore, miR-203a-3p mimics or FOXP1 knock-down reversed the effect of increased AC003090.1 expression on OD and adipogenic differentiation of hBMSCs. Dual luciferase reporter assays showed that AC003090.1 can sponge miR-203a-3p, which targets FOXP1 in hBMSCs. LncRNA AC003090.1 promotes OD of hBMSCs by regulating the miR-203a-3p/FOXP1 axis.
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CiteScore
4.30
自引率
17.20%
发文量
145
审稿时长
2.3 months
期刊介绍: Information not localized
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