遗传 ›› 2017, Vol. 39 ›› Issue (9): 828-836.doi: 10.16288/j.yczz.16-403

• 研究报告 • 上一篇    下一篇

miR-101a靶向EZH2促进山羊骨骼肌卫星细胞的分化

李俊涛1(),赵薇1,李丹丹1,冯静2,巴贵2,宋天增2,张红平1()   

  1. 1 四川农业大学动物遗传育种研究所，成都 611130
    2 西藏农牧科学院畜牧兽医研究所，拉萨 850009
  • 收稿日期:2017-02-22 修回日期:2017-04-25 出版日期:2017-09-20 发布日期:2017-10-21
  • 作者简介:李俊涛,硕士研究生,专业方向：动物遗传育种。E-mail: 1373721148@qq.com
  • 基金资助:
    四川省科技支撑计划项目(2016NYZ0045);西藏自治区财政项目(2015, Z2016B01N04-06)

miR-101a targeting EZH2 promotes the differentiation of goat skeletal muscle satellite cells

Juntao Li1(),Wei Zhao1,Dandan Li1,Jing Feng2,Gui Ba2,Tianzeng Song2,Hongping Zhang1()   

  1. 1 Institute of Animal Genetics and Breeding, Sichuan Agricultural University, Chengdu 611130, China
    2 Institute of Animal Science, Tibet Academy of Agricultural and Animal Husbandry Sciences, Lhasa 850009, China
  • Received:2017-02-22 Revised:2017-04-25 Online:2017-09-20 Published:2017-10-21
  • Supported by:
    the Science and Technology Program of Sichuan Province(2016NYZ0045);Tibet Autonomous Region Science and Technology Major Project(2015, Z2016B01N04-06)

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摘要：

本实验室前期研究发现,miR-101a对山羊骨骼肌卫星细胞(skeletal muscle satellite cells, SMSCs)分化有促进作用,但其具体作用机制并不清楚。本研究利用PicTar、TargetScan和miRanda软件在线预测miR-101a的靶基因,并通过双荧光素酶报告基因进行实验验证;检测了山羊SMSCs分化不同时期miR-101a和靶基因的表达关系,同时分析了超表达和抑制miR-101a对靶基因表达水平的影响。结果证实,zeste增强子同源物2(enhancer of zeste homologue 2, EZH2)基因mRNA的3°UTR具有miR-101a结合位点,是miR-101a的一个靶基因。在SMSCs分化过程中,随着miR-101a表达水平的升高,EZH2的表达在mRNA和蛋白水平均下调。抑制miR-101a后,EZH2的表达极显著升高(P<0.01),但是在超表达miR-101a条件下,EZH2表达变化在mRNA和蛋白水平均不显著(P>0.05)。以上研究结果表明,miR-101a能通过抑制EZH2的表达来促进山羊SMSCs分化,为进一步阐明miR-101a对SMSCs的调控机制提供了理论依据。

关键词: miR-101a, 山羊, 骨骼肌卫星细胞, zeste增强子同源物2

Abstract:

miR-101a promotes the differentiation of goat skeletal muscle satellite cells (SMSCs), as we previously reported, but the underpinning mechanism remains to be illuminated. In this study, we predicted the target gene of miR-101a by employing online softwares PicTar, TargetScan and miRanda, and found that enhancer of zeste homologue 2 (EZH2) was targeted by miR-101a. Further we identified that EZH2 contained miR-101a binding sites at its 3'UTR by using the dual-luciferase reporter assay system. In addition, we showed that during SMSC differentiation, the downregulated levels of EZH2 mRNA and protein were accompanied by increasing miR-101a expression via qRT-PCR and Western blot. Additionally, the expression of EZH2 significantly increased (P<0.01) when miR-101a was suppressed, whereas overexpressing miR-101a almost had no effect on EZH2 expression (P>0.05). These data demonstrated that miR-101a promotes SMSC differentiation directly through EZH2, which provides a theoretical reference for further elucidating the mechanism of miR-101a in SMSC differentiation.

Key words: miR-101a, goat, skeletal muscle satellite cells (SMSCs), enhancer of zeste homologue 2(EZH2)