A Novel Circular RNA Generated by FGFR2 Gene Promotes Myoblast Proliferation and Differentiation by Sponging miR-133a-5p and miR-29b-1-5p.
A Novel Circular RNA Generated by FGFR2 Gene Promotes Myoblast Proliferation and Differentiation by Sponging miR-133a-5p and miR-29b-1-5p.
复制标题
FGFR2 基因产生的新型环状 RNA 通过海绵 miR-133a-5p 和 miR-29b-1-5p 促进成肌细胞增殖和分化
作者：
Chen X;Ouyang H;Wang Z;Chen B;Nie Q
众所周知，成纤维细胞生长因子受体2（FGFR 2）与其配体成纤维细胞生长因子（FGF）相互作用，从而对细胞增殖和分化发挥生物学功能。在这项研究中，我们首次报道了FGFR 2基因可以产生circFGFR 2的环状RNA，其通过海绵状miRNA调节骨骼肌发育。在我们前期的环状RNA测序研究中，我们发现由FGFR 2基因外显子3-6产生的circFGFR 2在鸡胚骨骼肌发育过程中差异表达。本研究旨在揭示circFGFR 2影响鸡骨骼肌发育的真实的机制。在这项研究中，通过流式细胞术分析细胞周期和5-乙炔基-2 ′-脱氧尿苷（EdU）测定来分析细胞增殖。通过分析分化标记基因的表达和肌球蛋白重链（MyHC）免疫荧光来确定细胞分化。流式细胞仪细胞周期分析和EdU检测结果显示，过表达circFGFR 2促进了成肌细胞和QM-7细胞的增殖，而siRNA敲低circFGFR 2则抑制了这两种细胞的增殖。同时，过表达circFGFR 2可促进成肌细胞中肌细胞分化因子1（myogenic differentiation 1，MYOD）、肌细胞生成素（myogenin，MYOG）的表达和肌管的形成，而敲低circFGFR 2则相反。荧光素酶报告基因检测和生物素偶联的miRNA pull down检测结果进一步表明，circFGFR 2可直接靶向miR-133 a-5 p的两个结合位点和miR-29 b-1- 5 p的一个结合位点，进一步抑制这两种miRNA的表达和活性。此外，我们证明了miR-133 a-5 p和miR-29 b-1- 5 p都抑制成肌细胞增殖和分化，而circFGFR 2可以消除这两种miRNA的抑制作用，正如救援实验所表明的那样。总之，我们的数据表明，一种新的circFGFR 2环状RNA可以通过海绵状的miR-133 a-5 p和miR-29 b-1- 5 p促进骨骼肌的增殖和分化。
It is well known that fibroblast growth factor receptor 2 (FGFR2) interacts with its ligand of fibroblast growth factor (FGF) therefore exerting biological functions on cell proliferation and differentiation. In this study, we first reported that the FGFR2 gene could generate a circular RNA of circFGFR2, which regulates skeletal muscle development by sponging miRNA. In our previous study of circular RNA sequencing, we found that circFGFR2, generated by exon 3–6 of FGFR2 gene, differentially expressed during chicken embryo skeletal muscle development. The purpose of this study was to reveal the real mechanism of how circFGFR2 affects skeletal muscle development in chicken. In this study, cell proliferation was analyzed by both flow cytometry analysis of the cell cycle and 5-ethynyl-2′-deoxyuridine (EdU) assays. Cell differentiation was determined by analysis of the expression of the differentiation marker gene and Myosin heavy chain (MyHC) immunofluorescence. The results of flow cytometry analysis of the cell cycle and EdU assays showed that, overexpression of circFGFR2 accelerated the proliferation of myoblast and QM-7 cells, whereas knockdown of circFGFR2 with siRNA reduced the proliferation of both cells. Meanwhile, overexpression of circFGFR2 accelerated the expression of myogenic differentiation 1 (MYOD), myogenin (MYOG) and the formation of myotubes, and knockdown of circFGFR2 showed contrary effects in myoblasts. Results of luciferase reporter assay and biotin-coupled miRNA pull down assay further showed that circFGFR2 could directly target two binding sites of miR-133a-5p and one binding site of miR-29b-1-5p, and further inhibited the expression and activity of these two miRNAs. In addition, we demonstrated that both miR-133a-5p and miR-29b-1-5p inhibited myoblast proliferation and differentiation, while circFGFR2 could eliminate the inhibition effects of the two miRNAs as indicated by rescue experiments. Altogether, our data revealed that a novel circular RNA of circFGFR2 could promote skeletal muscle proliferation and differentiation by sponging miR-133a-5p and miR-29b-1-5p.
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影响因子：
64.8
作者：
Baek, Daehyun;Villen, Judit;Shin, Chanseok;Camargo, Fernando D.;Gygi, Steven P.;Bartel, David P.
通讯作者：
Bartel, David P.
捕获microRNA结合的mRNA将肿瘤抑制器miR-34a识别为生长因子信号传导的调节剂。
影响因子：
4.5
作者：
Lal A;Thomas MP;Altschuler G;Navarro F;O'Day E;Li XL;Concepcion C;Han YC;Thiery J;Rajani DK;Deutsch A;Hofmann O;Ventura A;Hide W;Lieberman J
通讯作者：
Lieberman J
影响因子：
4.6
作者：
Abe N;Matsumoto K;Nishihara M;Nakano Y;Shibata A;Maruyama H;Shuto S;Matsuda A;Yoshida M;Ito Y;Abe H
通讯作者：
Abe H
影响因子：
4.1
作者：
Dudekulay, Dawood B.;Panda, Amaresh C.;Gorospe, Myriam
通讯作者：
Gorospe, Myriam
影响因子：
64.5
作者：
ARNBERG, AC;VANOMMEN, GJB;BORST, P
通讯作者：
BORST, P