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miR-122-5p靶向KDM2A对骨髓间充质干细胞成骨分化的影响
Effect of miR-122-5p targeting KDM2A on osteogenic differentiation of bone marrow mesenchymal stem cell
投稿时间:2023-08-10  
DOI:10.3969/j.issn.1672-5972.2023.05.001
中文关键词:  miR-122-5p  组蛋白去甲基化酶2A  骨髓间充质干细胞  成骨分化
英文关键词:miR-122-5p  KDM2A  Bone marrow mesenchymal stem cell  Osteogenic differentiation
基金项目:湖北省重点实验室开放项目(2021KFY021)
作者单位邮编
徐捷* 武汉大学人民医院骨科湖北 武汉430060 430060
彭昊武汉大学人民医院骨科湖北 武汉430060 430060
万龙彪武汉大学人民医院骨科湖北 武汉430060 430060
刘丰武汉大学人民医院骨科湖北 武汉430060 430060
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中文摘要:
      目的 探讨miR-122-5p靶向组蛋白去甲基化酶2A(histone demethylation protein 2A, KDM2A)对骨髓间充质干细胞成骨分化的影响。方法 体外培养人骨髓间充质干细胞(bone marrow mesenchymal stem cell, BMSC),利用qRT-PCR和Western blot方法检测miR-122-5p和KDM2A对BMSC成骨相关指标(OPN、OCN和RUNX2)表达情况的影响;茜素红染色和碱性磷酸酶染色检测miR-122-5p和KDM2A对BMSC钙沉积和碱性磷酸酶活性的影响;双荧光素酶实验验证miR-122-5p和KDM2A之间的结合关系;miR-122-5p inhibitor和si-KDM2A共转染研究两者对BMSC成骨分化的影响。结果 miR-122-5p mimic和si-KDM2A能够促进BMSC成骨相关基因的表达水平、钙结节沉积及碱性磷酸酶活性,miR-122-5p和KDM2A两者之间存在靶向结合关系,且miR-122-5p能够靶向抑制KDM2A表达,进而促进BMSC成骨相关基因的表达水平、钙结节沉积及碱性磷酸酶活性。结论 miR-122-5p靶向抑制KDM2A表达,进而促进BMSC成骨分化。
英文摘要:
      Objective To investigate the effect of miR-122-5p targeting histone demethylation protein 2A (KDM2A) on the osteogenic differentiation of bone marrow mesenchymal stem cell (BMSC).Methods In vitro culture of human BMSC was conducted to examine the effects of miR-122-5p and KDM2A on the expression of osteogenesis-related indicators (OPN, OCN and RUNX2) by qRT-PCR and Western blot analysis. Alizarin Red staining and alkaline phosphatase staining were utilized to assess the influence of miR-122-5p and KDM2A on BMSC calcium deposition and alkaline phosphatase activity. Furthermore, dual luciferase experiments were performed to verify the binding relationship between miR-122-5p and KDM2A. In addition, co-transfection of miR-122-5p inhibitor and si-KDM2A was performed to investigate their respective effects on the osteogenic differentiation of BMSC.Results The findings demonstrated that miR-122-5p mimic and si-KDM2A enhance the expression level of osteogenesis-related genes, calcium nodule deposition, and alkaline phosphatase activity in BMSC. Additionally, a targeted binding relationship between miR-122-5p and KDM2A was observed, indicating that miR-122-5p can inhibit the expression of KDM2A, thereby promoting the expression level of osteogenesis-related genes, calcium nodule deposition, and alkaline phosphatase activity in BMSC.Conclusion The study concludes that miR-122-5p targets and inhibits the expression of KDM2A, which ultimately promotes the osteogenic differentiation of BMSC.
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