摘要
MicroRNAs(miRNAs)是一类小非编码RNA,近年研究发现其在骨骼肌发育调控中发挥重要作用.为探明miR-143-3p在C2C12成肌细胞分化中的调控作用,采用real-time PCR检测了miR-143-3p在小鼠各组织及C2C12成肌细胞分化过程中的表达;使用miR-143-3p的模拟物和特异性抑制剂分别处理细胞,采用real-time PCR和Western印迹分别检测成肌因子MyoG和成肌标志基因MyHC mRNA和蛋白水平的变化;用免疫荧光染色的方法观察肌管的形成.结果显示,miR-143-3p在小鼠各组织中均有表达,并且随着细胞分化表达量逐渐增加;C2C12成肌细胞过表达miR-143-3p,与对照组相比,成肌调控因子MyoG和成肌标志基因MyHC的mRNA和蛋白表达均显著升高,肌管数量明显增多;抑制剂处理结果显示,细胞分化被显著抑制.检测miR-143-3p对MyHC各亚型表达的影响发现,miR-143-3p表达的变化并不直接影响MyHC各亚型的表达.以上结果说明,miR-143-3p在骨骼肌和成肌细胞中均有表达,能够促进C2C12成肌细胞分化,但并不直接调控MyHCs的表达.
MicroRNAs （miRNAs） are small non-coding RNA that play important roles in skeletal muscle development. To explore the function of miR-143-3p in the differentiation of C2C12 myoblasts, we detected miR-143-3p levels by real-time PCR in different mouse tissues, as well as C2C12 myoblasts during myogenesis. After the trasfection of miR-143-3p mimics and inhibitor in C2C12 myoblasts, the expression of myogenic regulatory factor MyoG and myogenic marker gene MyHC were detected by real- time PCR and Western blotting. The myotubule formation was detected by immunofluorescent staining. The results showed that miR-143-3p was ubiquitously expressed in various tissues and was upregulated during cell differentiation. The differentiation of C2C12 myoblasts was promoted with miR-143-3p overexpression as significant upregulation of MyoG and MyHC, and increased number of myotubules. The inhibitor of miR-143-3p significantly repressed cell differentiation. Interestingly,the transfection of miR- 143-3p mimics had little effect on the expression of MyHCs. Our data suggested that miR-143-3p might be involved during the myogeneis of C2C12 myoblasts,but not directly impact MyHC expression.
出处
《中国生物化学与分子生物学报》
CAS
CSCD
北大核心
2013年第6期569-577,共9页
Chinese Journal of Biochemistry and Molecular Biology
基金
国家自然科学基金(No.31072014)
西北农林科技大学"创新团队建设计划"~~
