Ge Guihua, Yang Dongli, Tan Ya, Chen Ying, Jiang Dongmei, Jiang Anan, Li Qiang, Liu Yihui, Zhong Zhijun, Li Xuewei, Zhang Shunhua, Zhu Li
a College of Animal Science & Technology , Sichuan Agricultural University , Chengdu , Sichuan , China.
b Luzhou Animal Husbandry Station , Luzhou , Sichuan , China.
Biosci Biotechnol Biochem. 2019 Feb;83(2):291-299. doi: 10.1080/09168451.2018.1533805. Epub 2018 Oct 18.
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.
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。