| 引用本文： |
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秦兵莲,沈君颜,陈嘉瑜,等.miR-29b调节TET蛋白对脂肪来源的间充质干细胞成骨分化的影响[J].同济大学学报（医学版）,2017,38(5):7-12. [点击复制]
- QIN Bing-lian,SHEN Jun-yan1,CHEN Jia-yu,et al.MiR-29b affects osteogenic differentiation of adipocyte-derived mesenchymal stem cells by modulating TET protein[J].同济大学学报（医学版）,2017,38(5):7-12. [点击复制]
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| 摘要: |
| 目的 探讨在干细胞定向分化以及胚胎早期发育中，miRNA-29b对DNA去甲基化酶TET蛋白功能的调控机制。方法采用生物信息学的方法结合双荧光素酶报告基因检测进行体外验证获得靶miRNAs；通过受精卵显微注射进行体内验证；对脂肪来源的间充质干细胞（adipose-derived stromal cells， ASCs）定向分化，确认靶miRNAs对Tets基因的影响。结果小鼠Tet1、Tet2、Tet3基因都是miR-29abc的靶基因，且miR-29b抑制效率最高（P<0.05）；通过CRISPR/Cas9技术突变，miR-29b能增强Tets的表达（P<0.05），并能显著促进ASCs向成骨细胞分化的效率（P<0.05）。结论miR-29b可以下调Tets表达，并且下调miR-29b能显著促进间充质干细胞向成骨细胞分化的效率。 |
| 关键词: miR-29b Tets 间充质干细胞 成骨细胞分化 |
| DOI：10.16118/j.1008-0392.2017.05.002 |
| 投稿时间：2017-06-08 |
| 基金项目:国家重点基础研究发展(973)计划（2012CB966903）；国家自然科学基金（31671539） |
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| MiR-29b affects osteogenic differentiation of adipocyte-derived mesenchymal stem cells by modulating TET protein |
| QIN Bing-lian,SHEN Jun-yan1,CHEN Jia-yu,LIU Hai-liang |
| (Dept. of Regenerative Medicine, School of Medicine, Tongji University, Shanghai 200092, China;School of Life Science and Technology, Tongji University, Shanghai 200092, China;Translational Center for Stem Cell Research, School of Medicine, Tongji Hospital, Shanghai 200065, China) |
| Abstract: |
| Objective To investigate the mechanism of miRNA-29b regulating the function of DNA demethylase TET protein in stem cells during directional differentiation and early embryonic development. MethodsIn vitro validation of target miRNAs was performed by bioinformatics methods combined with double luciferase reporter assay and in vivo validation was performed by fertilized egg microinjection, and then the influence of target miRNAs on Tets genes in directional differentiation of adipose-derived stromal cells (ASCs) was identified. ResultsThe three mouse Tets genes were the target genes of miR-29abc, and the miR-29b showed the highest inhibition efficiency（P<0.05）. Mutation of miR-29b by using CRISPR/Cas9 technology enhanced the expression of Tets and significantly promoted the efficiency of osteogenesis differentiation of ASCs（P<0.05）. ConclusionMiR-29b can down-regulate the expression of Tets genes, and down-regulation of miR-29b can significantly promote the differentiation of adipocyte-derived mesenchymal stem cells into osteoblasts. |
| Key words: miR-29b Tets adipose-derived stromal cells osteogenic differentiation |
