Cardiovascular Center, Department of Internal Medicine, University of Michigan Medical Center, Ann Arbor, Michigan 48109.
Cardiovascular Center, Department of Internal Medicine, University of Michigan Medical Center, Ann Arbor, Michigan 48109; Institute of Reproductive and Stem Cell Engineering, Central South University, Changsha 410078, China.
J Biol Chem. 2010 Mar 26;285(13):9383-9389. doi: 10.1074/jbc.M109.095612. Epub 2010 Jan 29.
MicroRNAs (miRs) have been reported to play a critical role in muscle differentiation and function. The purpose of this study is to determine the role of miRs during smooth muscle cell (SMC) differentiation from embryonic stem cells (ESCs). MicroRNA profiling showed that miR-10a expression is steadily increased during in vitro differentiation of mouse ESCs into SMCs. Loss-of-function approaches using miR-10a inhibitors uncovered that miR-10a is a critical mediator for SMC lineage determination in our retinoic acid-induced ESC/SMC differentiation system. In addition, we have documented for the first time that histone deacetylase 4 is a novel target of miR-10a and mediates miR-10a function during ESC/SMC differentiation. To determine the molecular mechanism through which retinoic acid induced miR-10a expression, a consensus NF-kappaB element was identified in the miR-10a gene promoter by bioinformatics analysis, and chromatin immunoprecipitation assay confirmed that NF-kappaB could bind to this element. Finally, inhibition of NF-kappaB nuclear translocation repressed miR-10a expression and decreased SMC differentiation from ESCs. Our data demonstrate for the first time that miR-10a is a novel regulator in SMC differentiation from ESCs. These studies suggest that miR-10a may play important roles in vascular biology and have implications for the diagnosis and treatment of vascular diseases.
微小 RNA（miRs）已被报道在肌肉分化和功能中发挥关键作用。本研究旨在确定 miRs 在胚胎干细胞（ESCs）向平滑肌细胞（SMC）分化过程中的作用。miRNA 谱分析显示，miR-10a 的表达在体外小鼠 ESC 向 SMC 分化过程中稳步增加。使用 miR-10a 抑制剂的功能丧失方法表明，miR-10a 是我们的维甲酸诱导 ESC/SMC 分化系统中 SMC 谱系决定的关键介质。此外，我们首次记录到组蛋白去乙酰化酶 4 是 miR-10a 的一个新靶点，并在 ESC/SMC 分化过程中介导 miR-10a 功能。为了确定维甲酸诱导 miR-10a 表达的分子机制，通过生物信息学分析在 miR-10a 基因启动子中鉴定出一个公认的 NF-κB 元件，染色质免疫沉淀测定证实 NF-κB 可以结合到这个元件上。最后，抑制 NF-κB 核易位抑制 miR-10a 的表达并减少 ESC 向 SMC 的分化。我们的数据首次表明，miR-10a 是 ESC 向 SMC 分化的一种新的调节因子。这些研究表明，miR-10a 可能在血管生物学中发挥重要作用，并对血管疾病的诊断和治疗具有重要意义。
