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环状 RNA circMYL1 通过海绵吸附 miR-2400 抑制成肌细胞增殖并促进其分化。

Circular RNA circMYL1 Inhibit Proliferation and Promote Differentiation of Myoblasts by Sponging miR-2400.

机构信息

Key Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, College of Animal Science and Technology, Northwest A&F University, Yangling 712100, China.

Faculty of Veterinary Science, University of Nyala, Nyala 155, Sudan.

出版信息

Cells. 2021 Jan 16;10(1):176. doi: 10.3390/cells10010176.

DOI:10.3390/cells10010176
PMID:33467116
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7830797/
Abstract

Circular RNAs (circRNAs) are a class of endogenous non-coding RNAs (ncRNAs) involved in regulating skeletal muscle development by sponging miRNAs. In this study, we found that the circMYL1 expression was down-regulated during myoblast proliferation, while gradually up-regulated in myoblast differentiation. The potential role of circMYL1 was identified in the proliferation of bovine myoblast through mRNA and protein expression of proliferation marker genes (, , and ), cell counting kit-8 assay, flow cytometry analysis, and 5-ethynyl 2'-deoxyuridine (EdU) assay. Analysis of the expression of differentiation marker genes (, , and ) and immunofluorescence of Myosin heavy chain (MyHC) was used to assess cell differentiation. The proliferation analysis revealed that circMYL1 inhibited the proliferation of bovine primary myoblast. Furthermore, the differentiation analysis demonstrated that circMYL1 promoted the differentiation of bovine primary myoblast. The luciferase screening and RNA immunoprecipitation (RIP) assays found that circMYL1 could have interaction with miR-2400. Additionally, we demonstrated that miR-2400 promoted proliferation and inhibited differentiation of bovine primary myoblast, while circMYL1 may eliminate the effects of miR-2400, as showed by rescue experiments. Together, our results revealed that a novel circular RNA of circMYL1 could inhibit proliferation and promote differentiation of myoblast by sponging miR-2400.

摘要

环状 RNA（circRNAs）是一类内源性非编码 RNA（ncRNAs），通过海绵吸附 miRNAs 参与调节骨骼肌发育。在本研究中，我们发现 circMYL1 在成肌细胞增殖过程中表达下调，而在成肌细胞分化过程中逐渐上调。通过增殖标志物基因（、、和）的 mRNA 和蛋白表达、细胞计数试剂盒-8 分析、流式细胞术分析和 5-乙炔基-2'-脱氧尿苷（EdU）分析，鉴定了 circMYL1 在牛成肌细胞增殖中的潜在作用。通过分化标志物基因（、、和）的表达分析和肌球蛋白重链（MyHC）的免疫荧光染色来评估细胞分化。增殖分析表明 circMYL1 抑制牛原代成肌细胞的增殖。此外，分化分析表明 circMYL1 促进牛原代成肌细胞的分化。荧光素酶筛选和 RNA 免疫沉淀（RIP）实验发现 circMYL1 可以与 miR-2400 相互作用。此外，我们证明 miR-2400 促进牛原代成肌细胞的增殖并抑制其分化，而 circMYL1 可能通过海绵吸附 miR-2400 消除其作用，如通过挽救实验所示。总之，我们的结果表明，circMYL1 的一种新型环状 RNA 通过海绵吸附 miR-2400 抑制成肌细胞增殖并促进其分化。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/17ba/7830797/032aa8cfc51c/cells-10-00176-g009.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/17ba/7830797/1afec77375b2/cells-10-00176-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/17ba/7830797/f74c56597e66/cells-10-00176-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/17ba/7830797/b7543d2e7247/cells-10-00176-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/17ba/7830797/c11b980fc8f3/cells-10-00176-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/17ba/7830797/e50a48b30cf4/cells-10-00176-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/17ba/7830797/989e90325ffa/cells-10-00176-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/17ba/7830797/31d52e97a0ea/cells-10-00176-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/17ba/7830797/df6ad6e3f609/cells-10-00176-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/17ba/7830797/032aa8cfc51c/cells-10-00176-g009.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/17ba/7830797/1afec77375b2/cells-10-00176-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/17ba/7830797/f74c56597e66/cells-10-00176-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/17ba/7830797/b7543d2e7247/cells-10-00176-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/17ba/7830797/c11b980fc8f3/cells-10-00176-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/17ba/7830797/e50a48b30cf4/cells-10-00176-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/17ba/7830797/989e90325ffa/cells-10-00176-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/17ba/7830797/31d52e97a0ea/cells-10-00176-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/17ba/7830797/df6ad6e3f609/cells-10-00176-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/17ba/7830797/032aa8cfc51c/cells-10-00176-g009.jpg

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