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微小RNA-10b-5p调控C2C12成肌细胞的增殖与分化。

miR-10b-5p Regulates C2C12 Myoblasts Proliferation and Differentiation.

作者信息

Ge Guihua, Yang Dongli, Tan Ya, Chen Ying, Jiang Dongmei, Jiang Anan, Li Qiang, Liu Yihui, Zhong Zhijun, Li Xuewei, Zhang Shunhua, Zhu Li

机构信息

a College of Animal Science & Technology , Sichuan Agricultural University , Chengdu , Sichuan , China.

b Luzhou Animal Husbandry Station , Luzhou , Sichuan , China.

出版信息

Biosci Biotechnol Biochem. 2019 Feb;83(2):291-299. doi: 10.1080/09168451.2018.1533805. Epub 2018 Oct 18.

Abstract

The development of skeletal muscle is a complex process including myoblasts proliferation and differentiation. MicroRNAs (miRNAs) are small non-coding RNAs that regulate gene expression at post-transcriptional level. Increasing evidences indicate that miRNAs are important regulators in myogenic processes. Here, we reported that the expression of miR-10b-5p steadily decreased during myoblasts proliferation, but significantly increased during myoblasts differentiation. The over-expression of miR-10b-5p promoted myoblasts proliferation and blunted myofiber formation in C2C12 cells, while miR-10b-5p down-regulation showed an opposite result. At the same time, we observed that the down-regulation of nuclear factor of activated T-cells 5 (NFAT5) repressed the differentiation of C2C12 cells, and interestingly, miR-10b-5p could suppress NFAT5 expression. Luciferase activity assays confirmed that miR-10b-5p directly target the 3'-untranslated region (3'-UTR) of NFAT5. Overall, we proposed here a novel insight that miR-10b-5p regulates the proliferation and differentiation of C2C12 myoblasts, and the impact on myogenic differentiation is partly through targeting NFAT5. Abbreviations: NFAT5: nuclear factor of activated T-cells 5; Cyclin B: cycle protein B; Cyclin D1: cycle protein D1; Cyclin E: cycle protein E; CDK4: cyclin-dependent kinase 4; MyoD: myogenic differentiation antigen; MyoG: myogenin; Myf5: myogenic factor 5; MRF4: myogenic regulatory factor 4; MyHC: myosin heavy chain; AQP5: aquaporin-5; CACNA1C: calcium voltage-gated channel subunit alpha1 C; SRF: serum response factor; Pax7: paired box 7; KLF4: Kruppel-like factor 4; 3'-UTR: 3'-untranslated region; GM: growth medium; DM: differentiation medium.

摘要

骨骼肌的发育是一个复杂的过程,包括成肌细胞的增殖和分化。微小RNA(miRNA)是一类小的非编码RNA,在转录后水平调节基因表达。越来越多的证据表明,miRNA是成肌过程中的重要调节因子。在此,我们报道miR-10b-5p的表达在成肌细胞增殖过程中稳步下降,但在成肌细胞分化过程中显著增加。miR-10b-5p的过表达促进了C2C12细胞中成肌细胞的增殖并抑制了肌纤维的形成,而miR-10b-5p的下调则显示出相反的结果。同时,我们观察到活化T细胞核因子5(NFAT5)的下调抑制了C2C12细胞的分化,有趣的是,miR-10b-5p可以抑制NFAT5的表达。荧光素酶活性测定证实miR-10b-5p直接靶向NFAT5的3'-非翻译区(3'-UTR)。总体而言,我们在此提出了一个新的见解,即miR-10b-5p调节C2C12成肌细胞的增殖和分化,并且对成肌分化的影响部分是通过靶向NFAT5实现的。缩写:NFAT5:活化T细胞核因子5;细胞周期蛋白B:周期蛋白B;细胞周期蛋白D1:周期蛋白D1;细胞周期蛋白E:周期蛋白E;细胞周期蛋白依赖性激酶4:周期蛋白依赖性激酶4;生肌分化抗原:生肌分化抗原;肌细胞生成素:肌细胞生成素;生肌因子5:生肌因子5;生肌调节因子4:生肌调节因子4;肌球蛋白重链:肌球蛋白重链;水通道蛋白5:水通道蛋白-5;钙电压门控通道亚基α1C:钙电压门控通道亚基α1C;血清反应因子:血清反应因子;配对盒7:配对盒7;Kruppel样因子4:Kruppel样因子4;3'-UTR:3'-非翻译区;GM:生长培养基;DM:分化培养基。

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