A Novel Long Noncoding RNA, lncR-125b, Promotes the Differentiation of Goat Skeletal Muscle Satellite Cells by Sponging miR-125b
A Novel Long Noncoding RNA, lncR-125b, Promotes the Differentiation of Goat Skeletal Muscle Satellite Cells by Sponging miR-125b
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一种新型长非编码 RNA lncR-125b 通过海绵 miR-125b 促进山羊骨骼肌卫星细胞的分化
DOI：
10.3389/fgene.2019.01171
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发表时间：
2019
影响因子：
3.7
通讯作者：
Zhang Hongping
中科院分区：
文献类型：
--
作者：
Zhan Siyuan;Qin Chenyu;Li DanDan;Zhao Wei;Nie Lu;Cao Jiaxue;Guo Jiazhong;Zhong Tao;Wang Linjie;Li Li;Zhang Hongping
长链非编码RNA（lncRNA）已成为骨骼肌发生的重要调控因子，但肌发生相关lncRNA的研究很少，对它们的调控机制的认识也很有限，尤其是在山羊中。在这里，我们发现了一种新的lncRNA，TCONS_00006810（命名为lncR-125 b），来自我们之前的胎儿lncRNA测序数据，（妊娠45、60和105天，每个时间点三个生物学重复）和产后（出生后3天，n = 3）山羊骨骼肌，并发现其在山羊骨骼肌中高度表达，并在骨骼肌卫星细胞（SMSC）分化过程中逐渐上调。值得注意的是，lncR-125 b的过表达加速了肌细胞分化1（MyoD 1）和肌细胞生成素（MyoG）的表达，以及肌管的形成，而lncR-125 b的敲低在SMSCs中显示出相反的效果。双荧光素酶检测和实时定量聚合酶链反应的结果显示，lncR-125 b作为miR-125 b的分子海绵，胰岛素样生长因子2（IGF 2），骨骼肌生成的关键调节因子，是miR-125 b的直接靶基因。进一步的分析表明，lncR-125 b负调控SMSCs中miR-125 b的表达，并正调控IGF 2的表达。在机制上，lncR-125 b通过充当miR-125 b的竞争性内源RNA（ceRNA）来控制IGF 2表达，从而促进SMSC分化。这些研究结果确定lncR-125 b作为一种新的非编码调节肌细胞分化和骨骼肌发育的山羊。
Long noncoding RNAs (lncRNAs) have emerged as essential regulators of skeletal myogenesis, but few myogenesis-associated lncRNAs have been identified and our understanding of their regulatory mechanisms remains limited, particularly in goat. Here, we identified a novel lncRNA, TCONS_00006810 (named lncR-125b), from our previous lncRNA sequencing data on fetal (45, 60, and 105 days of gestation, three biological replicates for each point) and postnatal (3 days after birth, n = 3) goat skeletal muscle, and found that it is highly expressed in skeletal muscle and gradually upregulated during skeletal muscle satellite cell (SMSC) differentiation in goat. Notably, overexpression of lncR-125b accelerated the expression of myogenic differentiation 1 (MyoD 1) and myogenin (MyoG), and the formation of myotubes, and knockdown of lncR-125b showed opposite effects in SMSCs. Results of dual-luciferase assay and quantitative real-time polymerase chain reaction revealed that lncR-125b acts as a molecular sponge for miR-125b and that insulin-like growth factor 2 (IGF2), a critical regulator of skeletal myogenesis, is a direct target gene of miR-125b. Further analyses showed that lncR-125b negatively regulates miR-125b expression and positively regulates IGF2 expression in SMSCs. Mechanistically, lncR-125b promotes SMSC differentiation by functioning as a competing endogenous RNA (ceRNA) for miR-125b to control IGF2 expression. These findings identify lncR-125b as a novel noncoding regulator of muscle cell differentiation and skeletal muscle development in goat.