Song Chengchuang, Yang Jiameng, Jiang Rui, Yang Zhaoxin, Li Hui, Huang Yongzhen, Lan Xianyong, Lei Chuzhao, Ma Yun, Qi Xinglei, Chen Hong
Key Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, College of Animal Science and Technology, Northwest A&F University, Yangling, Shaanxi, China.
Institute for Conservation and Utilization of Agro-Bioresources in Dabie Mountains, College of Life Sciences, Xinyang Normal University, Xinyang, Henan, China.
J Cell Physiol. 2019 Sep;234(9):15742-15750. doi: 10.1002/jcp.28232. Epub 2019 Feb 22.
Skeletal muscle development is regulated by a series of regulatory factors, and also including noncoding RNA, especially microRNAs (miRNAs). Recently, miR-148a has been found to be involved in murine C2C12 differentiation by targeting ROCK1. However, the function of miR-148a-3p for the proliferation and apoptosis of bovine muscle cells has not been determined. In this study, we found that miR-148a-3p was highly expressed in fetal bovine skeletal muscle and exhibited a decreasing trend in muscle cells during its growth phase. Functional studies indicated that gain of miR-148a-3p inhibited the proliferation of bovine muscle cells and promoted apoptosis. Conversely, interference with miR-148a-3p inhibitor promoted muscle cell proliferation and inhibited its apoptosis. Mechanistically, KLF6 was confirmed as a new potential target gene of miR-148a-3p by TargetScan software prediction and the dual-luciferase assay verification. Additionally, after a gain or loss of KLF6, the function of KLF6 for muscle cell proliferation and apoptosis was opposite to that of miR-148a-3p. Collectively, these findings proposed a novel avenue whereby miR-148a-3p impeded bovine myoblast cell proliferation and promoted apoptosis through the posttranscriptional downregulation of KLF6.
骨骼肌发育受一系列调控因子调节，其中还包括非编码RNA，尤其是微小RNA（miRNA）。最近发现，miR-148a通过靶向ROCK1参与小鼠C2C12细胞分化。然而，miR-148a-3p对牛肌肉细胞增殖和凋亡的作用尚未确定。在本研究中，我们发现miR-148a-3p在胎牛骨骼肌中高表达，在肌肉细胞生长阶段呈下降趋势。功能研究表明，过表达miR-148a-3p可抑制牛肌肉细胞增殖并促进凋亡。相反，干扰miR-148a-3p抑制剂可促进肌肉细胞增殖并抑制其凋亡。机制上，通过TargetScan软件预测和双荧光素酶报告基因实验验证，KLF6被确认为miR-148a-3p的一个新的潜在靶基因。此外，KLF6表达量增减后，其对肌肉细胞增殖和凋亡的作用与miR-148a-3p相反。总之，这些发现提出了一条新途径，即miR-148a-3p通过转录后下调KLF6来阻碍牛成肌细胞增殖并促进凋亡。
