Song Chengchuang, Yang Zhaoxin, Jiang Rui, Cheng Jie, Yue Binglin, Wang Jian, Sun Xiaomei, Huang Yongzhen, Lan Xianyong, Lei Chuzhao, Chen Hong
Key Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, College of Animal Science and Technology, Northwest A&F University, Yangling 712100, China; Institute of Cellular and Molecular Biology, Jiangsu Normal University, Xuzhou 221116, China.
Key Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, College of Animal Science and Technology, Northwest A&F University, Yangling 712100, China.
Mol Ther Nucleic Acids. 2020 Sep 4;21:874-884. doi: 10.1016/j.omtn.2020.07.002. Epub 2020 Jul 10.
The role of long non-coding RNA (lncRNA) in the regulation of bovine skeletal muscle development remains poorly understood. The present study investigated the function and regulatory mechanism of a novel lncRNA, insulin-like growth factor 2 antisense transcript (IGF2 AS), in bovine myoblast proliferation and differentiation. Gain or loss of IGF2 AS was performed using an expression plasmid or small interfering RNA (siRNA), respectively. Bovine myoblasts were used to investigate the biological function and mechanisms of IGF2 AS in vitro. Results were conjointly analyzed by celluar and molecular biology experiments as well as bioinformatics. Functionally, IGF2 AS could promote proliferation and differentiation of bovine myoblasts. The preliminary mechanism suggests, on the one hand, that IGF2 AS could complement the IGF2 gene intron region and affect the stability and expression of IGF2 mRNA. On the other hand, RNA pull-down and immunoprecipitation assays demonstrated that IGF2 AS could directly bind to the interleukin enhancer binding factor 3 (ILF3) protein and maybe partly though it to regulate myogenesis. In conclusion, the novel identified lncRNA IGF2 AS promoted proliferation and differentiation of bovine myoblasts through various pathways.
长链非编码RNA(lncRNA)在牛骨骼肌发育调控中的作用仍知之甚少。本研究探讨了一种新型lncRNA,即胰岛素样生长因子2反义转录本(IGF2 AS)在牛成肌细胞增殖和分化中的功能及调控机制。分别使用表达质粒或小干扰RNA(siRNA)进行IGF2 AS的过表达或敲低。利用牛成肌细胞在体外研究IGF2 AS的生物学功能和机制。通过细胞和分子生物学实验以及生物信息学对结果进行联合分析。在功能上,IGF2 AS可促进牛成肌细胞的增殖和分化。初步机制表明,一方面,IGF2 AS可与IGF2基因内含子区域互补,影响IGF2 mRNA的稳定性和表达。另一方面,RNA下拉和免疫沉淀试验表明,IGF2 AS可直接与白细胞介素增强子结合因子3(ILF3)蛋白结合,并可能部分通过它来调节肌生成。总之,新鉴定的lncRNA IGF2 AS通过多种途径促进牛成肌细胞的增殖和分化。