为探究lncRNA对牛骨骼肌卫星细胞成肌分化过程的影响,选取前期测序结果中在牛肌卫星细胞成肌分化前后差异表达且表达丰度较高的lncRNA-HZ5进行成肌分化调控研究,利用q RT-PCR技术对其在牛骨骼肌卫星细胞成肌分化前后的表达变化进行检测,然后设计合成lncRNA-HZ5的si RNA,转染牛骨骼肌卫星细胞,抑制lncRNA-HZ5表达,之后将肌卫星细胞进行成肌分化诱导,通过肌管形成状态和MHC表达水平检测分析lncRNA-HZ5表达水平改变对肌卫星细胞成肌分化过程的影响。结果表明:lncRNA-HZ5在成肌分化前后的表达存在显著差异,干扰lncRNA-HZ5表达后的肌卫星细胞经成肌分化诱导,肌管数量及MHC的表达量均显著高于对照组,说明下调lncRNA-HZ5表达可以显著促进肌卫星细胞的成肌分化过程。本研究结果表明,lncRNA-HZ5可以负调控牛骨骼肌卫星细胞的成肌分化过程。
In order to investigate the effects of lncRNA in the myogenic differentiation process of bovine skeletal muscle satellite cells, a highly and differentially expressed lncRNA (lncRNA-HZ5) detected by RNA-seq technique was selected for further research. The expression changes of lncRNA-HZ5 during bovine muscle satellite ceils myogenic differentiation process were identified by qRT-PCR. The expression of lncRNA-HZ5 in muscle satellite cells were down regulated by siRNA, then the cells were transferred to differentiation medium, the myogenic process were evaluated by myotube formation and MHC expression levels. The results showed that the expression of lncRNA-HZ5 were significantly changed during bovine muscle satellite cells myogenic differentiation process. The myotube number and MHC expression levels of siRNA transferred group were significantly higher than that of control group. It was found that inhibition of lncRNA-HZ5 expression using siRNA can promoting the myogenic differentiation process of bovine skeletal muscle satellite cells.