Yang Li, Luo Hai-Jing, Gong Zhi-An, Zhang Wen-Tian, Cui Jing-Xuan, Fu Xue-Peng, Zhang Wei-Wei
Department of Life Science and Agroforestry, Qiqihar University, No. 42 Wenhua Street, Jianhua District, Qiqihar, 161000, PR China.
J Muscle Res Cell Motil. 2025 May 8. doi: 10.1007/s10974-025-09695-x.
microRNAs play a crucial role in the intricate process of muscle satellite cells proliferation and differentiation. Previous studies have demonstrated that miR-2400 can regulate bovine skeletal muscle satellite cell (MuSCs) proliferation, yet the underlying mechanism remains incompletely elucidated. In this study, we employed bioinformatics prediction and dual luciferase reporter assays to establish that miR-2400 directly targets the 3' untranslated regions (UTRs) of melanoma antigen family D1 (MAGED1) mRNA, thereby suppressing its expression. To ascertain whether miR-2400 affects the proliferation of MuSCs through MAGED1, we constructed the MAGED1 interference vector using RNA interference technology (RNAi) and assessed changes in MuSCs proliferation subsequent to MAGED1 interference. The experimental data indicate that the cell viability and the rate of EdU-positive cells of MuSCs were increased after interference with MAGED1. The proportion of S-phase cells and the expression level of cell cycle-associated proteins CCND2 and CCNB1 increased. These findings align with miR-2400's role in promoting cell proliferation and suggest that miR-2400 exerts its effects by directly targeting MAGED1.
微小RNA在肌肉卫星细胞增殖和分化的复杂过程中发挥着关键作用。先前的研究表明，miR - 2400可以调节牛骨骼肌卫星细胞（MuSCs）的增殖，但其潜在机制仍未完全阐明。在本研究中，我们采用生物信息学预测和双荧光素酶报告基因检测，确定miR - 2400直接靶向黑色素瘤抗原家族D1（MAGED1）mRNA的3'非翻译区（UTR），从而抑制其表达。为了确定miR - 2400是否通过MAGED1影响MuSCs的增殖，我们使用RNA干扰技术（RNAi）构建了MAGED1干扰载体，并评估了MAGED1干扰后MuSCs增殖的变化。实验数据表明，干扰MAGED1后，MuSCs的细胞活力和EdU阳性细胞率增加。S期细胞比例以及细胞周期相关蛋白CCND2和CCNB1的表达水平升高。这些发现与miR - 2400在促进细胞增殖中的作用一致，并表明miR - 2400通过直接靶向MAGED1发挥其作用。
