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

MiR-29b affects osteogenic differentiation of adipocyte-derived mesenchymal stem cells by modulating TET protein

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。

ObjectiveTo investigate the mechanism of miRNA29b 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 miR29abc,and the miR29b showed the highest inhibition efficiency(P<005).Mutation of miR29b by using CRISPR/Cas9technology enhanced the expression of Tets and significantly promoted the efficiency of osteogenesis differentiation of ASCs(P<005).ConclusionMiR29b can down regulate the expression of Tets genes,and down regulation of miR29b can significantly promote the differentiation of adipocyte derived mesenchymal stem cells into osteoblasts.