microRNA-27b suppresses mouse MSC migration to the liver by targeting SDF-1α in vitro
microRNA-27b suppresses mouse MSC migration to the liver by targeting SDF-1α in vitro
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DOI：
10.1016/j.bbrc.2012.04.027
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发表时间：
2012-05-04
影响因子：
3.1
通讯作者：
Yang, Shi-Ming
中科院分区：
文献类型：
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作者：
Lu, Mu-Han;Li, Chang-Zhu;Yang, Shi-Ming
SDF-1/CXCR4轴在诱导干细胞动员进入循环、将干细胞归巢到损伤部位以及干细胞参与肝组织再生方面是至关重要的。在这项研究中，我们深入了解了miRNAs调控SDF-1α表达的分子机制。利用微阵列和生物信息学方法，我们鉴定了六个在受损肝组织(肝损伤后21天)与正常C57BL/6小鼠肝组织相比表达差异的miRNAs，并通过qPCR进一步证实了这些观察结果；其他研究人员鉴定的miR-23a也被纳入进行比较。我们发现miR-23a、miR-27a和miR-27b在受损肝脏中的表达显著低于正常肝脏(p<；0.05)。我们进一步利用荧光素酶报告基因分析和Western印迹分析证实miR-27b可以直接与SDF-1α的3‘非编码区相互作用来抑制SDF-1α蛋白的表达。此外，我们通过Transwell实验发现，miR-27b的过表达显著减少了原代培养的CRCX4阳性的小鼠间充质干细胞(MMSCs)的定向迁移。这些结果提示miR-27b可能是受损小鼠肝脏干细胞生态位的独特标志，mir-27b通过直接与SDF-1α3‘UTR结合而下调SDF-1α的表达，从而抑制mMSCs的定向迁移。(C)2012 Elsevier Inc.保留所有权利。
The SDF-1/CXCR4 axis is critical for inducing stem cell mobilization into the circulation, for homing stem cells to the site of injury, and for stem cell participation in the regeneration of liver tissue. In this study, we have gained insight into the molecular mechanisms involved in regulating the expression of SDF-1 alpha by miRNAs. Using microarray and bioinformatics approaches, we identified six miRNAs with differential expression in damaged liver tissue (21 days after liver injury) compared to normal C57BL/6 murine liver tissue and further confirmed these observations by qPCR; miR-23a, which was identified by other researchers, was also included for comparative purposes. We found that miR-23a, miR-27a and miR-27b expression was significantly lower in the damaged liver than in the normal liver (p < 0.05). We further confirmed that miR-27b could directly interact with the 3'UTR of SDF-1 alpha to suppress SDF-1 alpha protein expression using a luciferase reporter assay and Western blot analysis. In addition, we found that the over-expression of miR-27b significantly reduced the directional migration of primary cultured CRCX4-positive murine mesenchymal stem cells (mMSCs) in vitro using a transwell assay. These results suggest that miR-27b may be a unique signature of the stem cell niche in the damaged mouse liver and that mir-27b can suppress the directional migration of mMSCs by down-regulating SDF-1 alpha expression by binding directly to the SDF-1 alpha 3'UTR. (C) 2012 Elsevier Inc. All rights reserved.