Key Laboratory of Agricultural Animal Genetics, Breeding, and Reproduction of the Ministry of Education, Key Laboratory of Swine Genetics and Breeding of the Ministry of Agriculture, College of Animal Science and Technology, Huazhong Agricultural University, Wuhan, Hubei, China.
National Engineering Research Center for Livestock, Huazhong Agricultural University, Wuhan, Hubei, China.
J Cell Physiol. 2024 Nov;239(11):e31359. doi: 10.1002/jcp.31359. Epub 2024 Jul 10.
Skeletal muscle constitutes the largest percentage of tissue in the animal body and plays a pivotal role in the development of normal life activities in the organism. However, the regulation mechanism of skeletal muscle growth and development remains largely unclear. This study investigated the effects of Ankrd1 on the proliferation and differentiation of C2C12 myoblasts. Here, we identified Ankrd1 as a potential regulator of muscle cell development, and found that Ankrd1 knockdown resulted in the proliferation ability decrease but the differentiation level increase of C2C12 cells. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyzes as well as RNA-seq results showed that Ankrd1 knockdown activated focal adhesion kinase (FAK)/F-actin signal pathway with most genes significantly enriched in this pathway upregulated. The integrin subunit Itga6 promoter activity is increased when Ankrd1 knockdown, as demonstrated by a dual-luciferase reporter assay. This study revealed the molecular mechanism by which Ankrd1 knockdown enhanced FAK phosphorylation activity through the alteration of integrin subunit levels, thus activating FAK/Rho-GTPase/F-actin signal pathway, eventually promoting myoblast differentiation. Our data suggested that Ankrd1 might serve as a potential regulator of muscle cell development. Our findings provide new insights into skeletal muscle growth and development and valuable references for further study of human muscle-related diseases.
骨骼肌构成了动物体内组织的最大比例，在生物体正常生命活动的发展中起着关键作用。然而，骨骼肌生长和发育的调节机制在很大程度上仍不清楚。本研究探讨了 Ankrd1 对 C2C12 成肌细胞增殖和分化的影响。在这里，我们确定 Ankrd1 是肌肉细胞发育的潜在调节剂，发现 Ankrd1 敲低导致 C2C12 细胞的增殖能力下降，但分化水平增加。基因本体论和京都基因与基因组百科全书通路富集分析以及 RNA-seq 结果表明，Ankrd1 敲低激活了粘着斑激酶（FAK）/F-肌动蛋白信号通路，大多数基因在此通路中显著上调。通过双荧光素酶报告基因检测证实，当 Ankrd1 敲低时，整合素亚基 Itga6 的启动子活性增加。本研究揭示了 Ankrd1 通过改变整合素亚基水平增强 FAK 磷酸化活性的分子机制，从而激活 FAK/Rho-GTPase/F-肌动蛋白信号通路，最终促进成肌细胞分化。我们的数据表明，Ankrd1 可能作为肌肉细胞发育的潜在调节剂。我们的研究结果为骨骼肌生长和发育提供了新的见解，并为进一步研究人类肌肉相关疾病提供了有价值的参考。
