环状 RNA TCF4 通过不依赖于 AGO2 结合抑制山羊骨骼肌卫星细胞的增殖和分化。

CircTCF4 Suppresses Proliferation and Differentiation of Goat Skeletal Muscle Satellite Cells Independent from AGO2 Binding.

机构信息

Farm Animal Genetic Resources Exploration and Innovation Key Laboratory of Sichuan Province, College of Animal Science and Technology, Sichuan Agricultural University, Chengdu 611130, China.

出版信息

Int J Mol Sci. 2022 Oct 25;23(21):12868. doi: 10.3390/ijms232112868.

DOI:10.3390/ijms232112868

PMID:36361663

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9657166/

Abstract

The proliferation and differentiation of mammalian skeletal muscle satellite cells (MuSCs) are highly complicated. Apart from the regulatory signaling cascade driven by the protein-coding genes, non-coding RNAs such as microRNAs (miRNA) and circular RNAs (circRNAs) play essential roles in this biological process. However, circRNA functions in MuSCs proliferation and differentiation remain largely to be elucidated. Here, we screened for an exonic circTCF4 based on our previous RNA-Seq data, specifically expressed during the development of the longest dorsal muscle in goats. Subsequently, the circular structure and whole sequence of circTCF4 were verified using Sanger sequencing. Besides, circTCF4 was spatiotemporally expressed in multiple tissues from goats but strikingly enriched in muscles. Furthermore, circTCF4 suppressed MuSCs proliferation and differentiation, independent of AGO2 binding. Finally, we conducted Poly(A) RNA-Seq using cells treated with small interfering RNA targeting circTCF4 and found that circTCF4 would affect multiple signaling pathways, including the insulin signaling pathway and AMPK signaling pathway related to muscle differentiation. Our results provide additional solid evidence for circRNA regulating skeletal muscle formation.

摘要

哺乳动物骨骼肌卫星细胞（MuSCs）的增殖和分化过程非常复杂。除了由蛋白质编码基因驱动的调控信号级联之外，非编码 RNA（如 microRNA（miRNA）和 circular RNA（circRNA））在这一生物学过程中也起着至关重要的作用。然而，circRNA 在 MuSCs 增殖和分化中的功能仍有待阐明。在这里，我们根据我们之前的 RNA-Seq 数据筛选出一种外显 circTCF4，该 circRNA 在山羊最长背肌发育过程中特异性表达。随后，使用 Sanger 测序验证了 circTCF4 的环状结构和全长序列。此外，circTCF4 在山羊的多种组织中具有时空表达特异性，但在肌肉中显著富集。此外，circTCF4 抑制 MuSCs 的增殖和分化，这一过程独立于 AGO2 结合。最后，我们使用针对 circTCF4 的小干扰 RNA 处理的细胞进行了 Poly(A) RNA-Seq，发现 circTCF4 会影响多种信号通路，包括与肌肉分化相关的胰岛素信号通路和 AMPK 信号通路。我们的研究结果为 circRNA 调节骨骼肌形成提供了额外的有力证据。