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目的探讨先天性纯红细胞再生障碍性贫血(DBA)相关microRNA的表达特征及其调控机制。方法利用morpholino技术建立核糖体蛋白基因Rps19表达不足的DBA斑马鱼模型和Rps19与p53同时表达不足的斑马鱼模型,利用新一代高通量测序(RNA-seq)技术,研究DBA斑马鱼模型的microRNA表达谱的特征及p53在其中的调控作用。结果 Rps19 MO的斑马鱼胚胎的循环血细胞明显减少,且尾部呈现出向腹侧弯曲的异常形态。而在p53敲低之后,斑马鱼的形态异常得到了一定程度的恢复。通过对microRNA表达谱的比较分析发现,在Rps19不足的斑马鱼胚胎中约1/4的microRNA表达量显著偏低,这些异常表达的microRNA可能影响到斑马鱼的正常发育过程。而在p53敲低之后,部分microRNA的表达恢复到正常水平。最终确定了47个在DBA发病过程中可能受p53调控的microRNA。结论 DBA斑马鱼模型的microRNA普遍低表达,其中部分microRNA受到p53的表达水平的调控。
Objective To investigate the expression and regulation of microRNAs associated with congenital pure erythrocyte aplasia (DBA). Methods The zebrafish model of DBA with under-expression of ribosomal protein gene Rps19 and the simultaneous expression of Rps19 and p53 were established by using morpholino technique. A new generation of high-throughput sequencing (RNA-seq) The characteristics of the expression profile and the role of p53 in its regulation. As a result, the number of circulating blood cells in the zebrafish embryos of Rps19 MO was significantly reduced, and the tail showed an abnormal morphology of ventral bending. After p53 knockdown, zebrafish morphological abnormalities have been restored to some extent. A comparative analysis of microRNA expression profiles revealed that about one-fourth of microRNAs expressed in zebrafish embryos deficient in Rps19 were significantly lower, and these abnormally expressed microRNAs might affect the normal development of zebrafish. After the knockdown of p53, the expression of some microRNAs returned to normal levels. Finally, 47 microRNAs that may be regulated by p53 during the pathogenesis of DBA were identified. Conclusion The microRNA of DBA zebrafish model is generally low expression, some of which are regulated by the expression level of p53.