College of Animal Science and Technology, Xinyang Agriculture and Forestry University, Xinyang, Henan 464000, P.R. China.
College of Animal Science and Technology, Northwest A&F University, Yangling, Shaanxi, 712100, P.R. China.
J Anim Sci. 2024 Jan 3;102. doi: 10.1093/jas/skae142.
Skeletal muscle is an important component of livestock and poultry organisms. The proliferation and differentiation of myoblasts are highly coordinated processes, which rely on the regulation of miRNA. MiRNAs are widely present in organisms and play roles in various biological processes, including cell proliferation, differentiation, and apoptosis. MiR-181d and miR-196a, identified as tumor suppressors, have been found to be involved in cell proliferation, apoptosis, directed differentiation, and cancer cell invasion. However, their role in beef cattle skeletal muscle metabolism remains unclear. In this study, we discovered that overexpression of bta-miR-181d and bta-miR-196a in Qinchuan cattle myoblasts inhibited proliferation and apoptosis while promoting myogenic differentiation through EDU staining, flow cytometry analysis, immunofluorescence staining, and Western blotting. RNA-seq analysis of differential gene expression revealed that after overexpression of bta-miR-181d and bta-miR-196a, the differentially expressed genes were mainly enriched in the PI3K-Akt and MAPK signaling pathways. Furthermore, the phosphorylation levels of key proteins p-AKT in the PI3K signaling pathway and p-MAPK in the MAPK signaling pathway were significantly decreased after overexpression of bta-miR-181d and bta-miR-196a. Overall, this study provides preliminary evidence that bta-miR-181d and bta-miR-196a may regulate proliferation, apoptosis, and differentiation processes in Qinchuan cattle myoblasts by affecting the phosphorylation status of key proteins in PI3K-Akt and MAPK-ERK signaling pathways.
骨骼肌是家畜和家禽机体的重要组成部分。肌细胞的增殖和分化是高度协调的过程,依赖于 miRNA 的调节。miRNA 广泛存在于生物体中,在细胞增殖、分化和凋亡等多种生物过程中发挥作用。miR-181d 和 miR-196a 作为肿瘤抑制因子,已被发现参与细胞增殖、凋亡、定向分化和癌细胞侵袭。然而,它们在肉牛骨骼肌代谢中的作用尚不清楚。在本研究中,我们发现 bta-miR-181d 和 bta-miR-196a 在秦川牛肌细胞中的过表达抑制了增殖和凋亡,同时通过 EDU 染色、流式细胞术分析、免疫荧光染色和 Western blot 促进了成肌分化。差异基因表达的 RNA-seq 分析表明,bta-miR-181d 和 bta-miR-196a 过表达后,差异表达基因主要富集在 PI3K-Akt 和 MAPK 信号通路中。此外,PI3K 信号通路中关键蛋白 p-AKT 和 MAPK 信号通路中 p-MAPK 的磷酸化水平在 bta-miR-181d 和 bta-miR-196a 过表达后显著降低。综上所述,本研究初步表明,bta-miR-181d 和 bta-miR-196a 可能通过影响 PI3K-Akt 和 MAPK-ERK 信号通路中关键蛋白的磷酸化状态,调节秦川牛肌细胞的增殖、凋亡和分化过程。