Li Hui, Wei Xuefeng, Yang Jiameng, Dong Dong, Hao Dan, Huang Yongzhen, Lan Xianyong, Plath Martin, Lei Chuzhao, Ma Yun, Lin Fengpeng, Bai Yueyu, Chen Hong
Shaanxi Key Laboratory of Molecular Biology for Agriculture, College of Animal Science and Technology, Northwest A&F University, Yangling, 712100 Shaanxi, China.
College of Life Sciences, Xinyang Normal University, Institute for Conservation and Utilization of Agro-Bioresources in Dabie Mountains, Xinyang, Henan 464000, PRC.
Mol Ther Nucleic Acids. 2018 Jun 1;11:272-283. doi: 10.1016/j.omtn.2018.02.012. Epub 2018 Mar 6.
Muscle development is regulated under a series of complicate processes, and non-coding RNAs, such as microRNAs (miRNAs) and circular RNAs (circRNAs), have been reported to play important roles in regulating myoblast proliferation and differentiation. We found that miR-107 expression was high in skeletal muscle of Qinchuan cattle. Overexpression of miR-107 inhibited bovine myoblasts differentiation and protected cells from apoptosis. Wnt3a was identified as a target of miR-107 by luciferase activity, real-time qPCR, and western blotting assays. Knockdown of Wnt3a inhibited bovine myoblasts differentiation and apoptosis, and this effect was similar to miR-107 overexpression. We also found circFGFR4 to promote myoblasts differentiation and to induce cell apoptosis. Via luciferase screening and RNA pull-down assays, circFGFR4 was observed to sponge miR-107. Overexpression of circFGFR4 increased the expression of Wnt3a, whereas this effect was abolished by miR-107. These results demonstrated that circFGFR4 binding miR-107 promotes cell differentiation via targeting Wnt3a in bovine primary myoblasts.
肌肉发育受一系列复杂过程调控，据报道，非编码RNA，如微小RNA（miRNA）和环状RNA（circRNA），在调节成肌细胞增殖和分化中发挥重要作用。我们发现，秦川牛骨骼肌中miR-107表达较高。miR-107的过表达抑制了牛成肌细胞的分化，并保护细胞免于凋亡。通过荧光素酶活性、实时定量PCR和蛋白质免疫印迹分析，Wnt3a被鉴定为miR-107的一个靶点。敲低Wnt3a抑制了牛成肌细胞的分化和凋亡，这种效应与miR-107过表达相似。我们还发现circFGFR4促进成肌细胞分化并诱导细胞凋亡。通过荧光素酶筛选和RNA下拉分析，观察到circFGFR4可吸附miR-107。circFGFR4的过表达增加了Wnt3a的表达，而miR-107消除了这种效应。这些结果表明，在牛原代成肌细胞中，circFGFR4结合miR-107通过靶向Wnt3a促进细胞分化。
