Wang Shan, Cao Xiukai, Ge Ling, Gu Yifei, Lv Xiaoyang, Getachew Tesfaye, Mwacharo Joram M, Haile Aynalem, Sun Wei
College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China.
Joint International Research Laboratory of Agriculture, Agri-Product Safety of Ministry of Education of China, Yangzhou University, Yangzhou 225009, China.
Animals (Basel). 2022 Jan 4;12(1):114. doi: 10.3390/ani12010114.
The growth and development of skeletal muscle require a series of regulatory factors. MiRNA is a non-coding RNA with a length of about 22 nt, which can inhibit the expression of mRNA and plays an important role in the growth and development of muscle cells. The role of miR-22-3p in C2C12 cells and porcine skeletal muscle has been reported, but it has not been verified in Hu sheep skeletal muscle. Through qPCR, CCK-8, EdU and cell cycle studies, we found that overexpression of miR-22-3p inhibited proliferation of skeletal muscle cells ( < 0.01). The results of qPCR and immunofluorescence showed that overexpression of miR-22-3p promoted differentiation of skeletal muscle cells ( < 0.01), while the results of inhibiting the expression of miR-22-3p were the opposite. These results suggested that miR-22-3p functions in growth and development of sheep skeletal muscle cells. Bioinformatic analysis with mirDIP, miRTargets, and RNAhybrid software suggested was the target of miR-22-3p, which was confirmed by dual-luciferase reporter system assay. is highly expressed in sheep skeletal muscle cells. Overexpression of was found to promote proliferation of skeletal muscle cells indicated by qPCR, CCK-8, EdU, and cell cycle studies ( < 0.01). The results of qPCR and immunofluorescence experiments proved that overexpression of inhibited differentiation of skeletal muscle cells ( < 0.01), while the results of interfering with siRNA were the opposite. These results indicate that miR-22-3p is involved in proliferation and differentiation of skeletal muscle cells by targeting .
骨骼肌的生长发育需要一系列调控因子。微小RNA（miRNA）是一种长度约为22个核苷酸的非编码RNA，它可抑制信使核糖核酸（mRNA）的表达，在肌肉细胞的生长发育中发挥重要作用。已有关于miR-22-3p在C2C12细胞和猪骨骼肌中作用的报道，但尚未在湖羊骨骼肌中得到验证。通过定量聚合酶链反应（qPCR）、细胞计数试剂盒-8（CCK-8）、5-乙炔基-2'-脱氧尿苷（EdU）和细胞周期研究，我们发现miR-22-3p过表达抑制骨骼肌细胞增殖（P<0.01）。qPCR和免疫荧光结果显示，miR-22-3p过表达促进骨骼肌细胞分化（P<0.01），而抑制miR-22-3p表达的结果则相反。这些结果表明，miR-22-3p在绵羊骨骼肌细胞的生长发育中发挥作用。使用mirDIP、miRTargets和RNAhybrid软件进行的生物信息学分析表明[某基因]是miR-22-3p的靶标，双荧光素酶报告系统检测证实了这一点。[某基因]在绵羊骨骼肌细胞中高表达。qPCR、CCK-8、EdU和细胞周期研究表明，[某基因]过表达促进骨骼肌细胞增殖（P<0.01）。qPCR和免疫荧光实验结果证明，[某基因]过表达抑制骨骼肌细胞分化（P<0.01），而用小干扰RNA（siRNA）干扰[某基因]的结果则相反。这些结果表明，miR-22-3p通过靶向[某基因]参与骨骼肌细胞的增殖和分化。
