College of Veterinary Medicine, Institute of Comparative Medicine, Yangzhou University, Yangzhou, PR China.
College of Animal Science and Technology, Yangzhou University, Yangzhou, PR China.
Anim Biotechnol. 2023 Nov;34(5):1727-1736. doi: 10.1080/10495398.2022.2044840. Epub 2022 Mar 9.
MicroRNAs are involved in a series of biological processes, such as proliferation, differentiation and apoptosis of primary myoblasts. The research group found that miR-214 is highly expressed in chicken primary myoblasts (CPMs), so we used miR-214 as a starting point to explore the biological function of miR-214 in skeletal muscle growth and development. In this experiment, CPMs were cultured in vitro; miR-214 was overexpressed in CPMs; and cell samples were collected for subsequent transcriptome sequencing (RNA-seq). After miR-214 overexpression, we identified 97 differentially expressed genes (DEGs), of which 21 DEGs were up-regulated and 76 DEGs were down-regulated. After bioinformatics analysis, these DEGs were found to be significantly enriched in myofibrils, muscle system processes, myofibril assembly and other biological processes related to muscle development. The significantly enriched KEGGs include focal adhesion and type II diabetes mellitus. The protein network of DEGs was drawn by STRING and Cytoscape software, and 5 DEGs were randomly selected to verify the sequencing results by real-time fluorescence quantification. CAV3 is not only an important node protein in the protein network but also a member of the focal adhesion signaling pathway. It is speculated that miR-214 may regulate muscle development through the focal adhesion signaling pathway.
微小 RNA 参与了一系列生物学过程,如原代肌细胞的增殖、分化和凋亡。研究小组发现 miR-214 在鸡原代肌细胞(CPM)中高度表达,因此我们以 miR-214 为起点,探索 miR-214 在骨骼肌生长和发育中的生物学功能。在本实验中,体外培养 CPM;在 CPM 中转染 miR-214 过表达载体;收集细胞样本进行后续转录组测序(RNA-seq)。miR-214 过表达后,我们鉴定出 97 个差异表达基因(DEGs),其中 21 个 DEGs 上调,76 个 DEGs 下调。通过生物信息学分析,这些 DEGs 被富集在肌原纤维、肌肉系统过程、肌原纤维组装等与肌肉发育相关的生物学过程中。显著富集的 KEGG 包括粘着斑和 2 型糖尿病。通过 STRING 和 Cytoscape 软件绘制 DEGs 的蛋白网络图,随机选取 5 个 DEGs 进行实时荧光定量验证测序结果。CAV3 不仅是蛋白网络图中的一个重要节点蛋白,也是粘着斑信号通路的成员。推测 miR-214 可能通过粘着斑信号通路调节肌肉发育。