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microRNA-181a 调控湖羊骨骼肌卫星细胞的增殖与分化并靶向 YAP1 基因

MicroRNA-181a Regulates the Proliferation and Differentiation of Hu Sheep Skeletal Muscle Satellite Cells and Targets the YAP1 Gene.

机构信息

College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China.

Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University, Yangzhou 225009, China.

出版信息

Genes (Basel). 2022 Mar 16;13(3):520. doi: 10.3390/genes13030520.

Abstract

MicroRNA (miRNA) is of great importance to muscle growth and development, including the regulation of the proliferation and differentiation of skeletal muscle satellite cells (SMSCs). In our research group's previous study, we found that miR-181a is differentially expressed in the longissimus dorsi muscle of Hu sheep at different stages. We speculated that miR-181a may participate in the growth and development process of Hu sheep. To understand the mechanism of miR-181a regulating the growth and development of Hu sheep skeletal muscle, we extracted skeletal muscle satellite cells from the longissimus dorsi muscle of 3-month-old Hu sheep fetuses and performed a series of experiments. Our results showed that miR-181a suppressed SMSCs' proliferation using QRT-PCR, Western blot, CCK-8, EDU, and Flow cytometry cycle tests. In addition, QRT-PCR, Western blot, and immunofluorescence indicated that miR-181a facilitated the differentiation of SMSCs. Then, we used dual-luciferase reporter gene detection, QRT-PCR, and Western blot to find that the Yes1-related transcription regulator (YAP1) is the target gene of miR-181a. Our study supplies a research basis for understanding the regulation mechanism of miR-181a on the growth of Hu sheep skeletal muscle.

摘要

微小 RNA(miRNA)对肌肉的生长和发育非常重要,包括调节骨骼肌卫星细胞(SMSCs)的增殖和分化。在我们研究小组之前的研究中,我们发现 miR-181a 在不同阶段的湖羊背最长肌中差异表达。我们推测 miR-181a 可能参与了湖羊的生长发育过程。为了了解 miR-181a 调节湖羊骨骼肌生长和发育的机制,我们从 3 月龄湖羊胎儿的背最长肌中提取了骨骼肌卫星细胞,并进行了一系列实验。我们的结果表明,miR-181a 通过 QRT-PCR、Western blot、CCK-8、EDU 和流式细胞术周期试验抑制 SMSCs 的增殖。此外,QRT-PCR、Western blot 和免疫荧光表明 miR-181a 促进了 SMSCs 的分化。然后,我们使用双荧光素酶报告基因检测、QRT-PCR 和 Western blot 发现 Yes1 相关转录调节剂(YAP1)是 miR-181a 的靶基因。我们的研究为了解 miR-181a 对湖羊骨骼肌生长的调节机制提供了研究基础。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9d3c/8949050/3c74847f2ee1/genes-13-00520-g001.jpg

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