College of Animal Science and Technology, Ningxia University, Yinchuan 750021, China; Key Laboratory of Ruminant Molecular Cell Breeding, Ningxia Hui Autonomous Region, Yinchuan 750021, China.
Institute of Animal Science, Ningxia Academy of Agriculture and Forestry Sciences, Yinchuan 750021, China.
Int J Biol Macromol. 2024 Mar;260(Pt 2):129643. doi: 10.1016/j.ijbiomac.2024.129643. Epub 2024 Jan 20.
The growth and development of bovine skeletal muscle and beef yield is closely intertwined. Our previous research found that forkhead box O1 (FOXO1) plays an important role in the regulation of beef muscle formation, but its specific mechanism is still unknown. In this study, we aimed to clarify the regulatory mechanism of FOXO1 in proliferation and differentiation of bovine skeletal muscle cells (BSMCs). The results showed that interfering with FOXO1 can promote proliferation and the cell G1/S phase of BSMCs by up-regulating the expression of PCNA, CDK1, CDK2, CCNA2, CCNB1, CCND1 and CCNE2. Besides, interfering with FOXO1 inhibited the apoptosis of BSMCs by up-regulating the expression of anti-apoptosis gene BCL2, while simultaneously down-regulating the expression of the pro-apoptosis genes BAD and BAX. Inversely, interfering with FOXO1 can promote the differentiation of BSMCs by up-regulating the expression of myogenic differentiation marker genes MYOD, MYOG, MYF5, MYF6 and MYHC. Furthermore, RNA-seq combined with western bolt, immunofluorescence and chromatin immunoprecipitation analysis showed that FOXO1 could regulate BSMCs differentiation process by influencing PI3K-Akt, Relaxin and TGF-beta signaling pathways, and target MYH3 for transcriptional inhibition. In conclusion, this study provides a basis for studying the role and molecular mechanism of FOXO1 in BSMCs.
牛骨骼肌的生长发育和牛肉产量密切相关。我们之前的研究发现叉头框 O1（FOXO1）在调节牛肉肌肉形成中起重要作用，但具体机制尚不清楚。在这项研究中，我们旨在阐明 FOXO1 在牛骨骼肌细胞（BSMC）增殖和分化中的调节机制。结果表明，干扰 FOXO1 可以通过上调 PCNA、CDK1、CDK2、CCNA2、CCNB1、CCND1 和 CCNE2 的表达来促进 BSMC 的增殖和细胞 G1/S 期。此外，干扰 FOXO1 通过上调抗凋亡基因 BCL2 的表达抑制 BSMC 的凋亡，同时下调促凋亡基因 BAD 和 BAX 的表达。相反，通过上调肌生成分化标记基因 MYOD、MYOG、MYF5、MYF6 和 MYHC 的表达，干扰 FOXO1 可以促进 BSMC 的分化。此外，RNA-seq 结合 western blot、免疫荧光和染色质免疫沉淀分析表明，FOXO1 可以通过影响 PI3K-Akt、松弛素和 TGF-β信号通路，以及靶向 MYH3 进行转录抑制，调节 BSMC 的分化过程。总之，本研究为研究 FOXO1 在 BSMC 中的作用和分子机制提供了依据。
