急性高温低氧胁迫下刺参体腔液细胞miRNA表达谱的构建和靶标基因的功能分析

Construction of miRNA Expression Profile in Coelomic Fluid Cells of Apostichopus japonicus and Functional Analysis of Their Target Genes Under Acute Heat Stress and Hypoxia Stress

为了探究刺参应对极端高温、低氧环境的内在分子调控机制,促进刺参养殖产业的可持续发展。本研究以刺参体腔液细胞作为研究对象,分别构建了高温、低氧下miRNA的差异表达谱并分析了差异表达miRNAs的靶基因功能。研究表明:高温组中共发掘了20个差异表达miRNA,其中11个上调,9个下调;低氧组中共发掘了10个差异表达miRNA,其中6个上调,4个下调。高温和低氧胁迫下共同显著差异表达的miRNA为:novel-23、novel-43、novel-45、novel-49、miR-200-3p、miR-71。以三代全长转录组为参考文库,利用miRanda软件对差异表达miRNA的靶基因进行预测,靶标基因KEGG富集分析显示,高温组中共显著富集到36条通路,低氧组中共显著富集到37条通路。其中磷脂酰肌醇激酶-丝氨酸/苏氨酸激酶信号通路(PI3K-Akt signaling pathway)、癌症通路(Pathways in cancer)、黏着斑(Focal adhesion)、丝裂原活化蛋白激酶信号通路(MAPK signaling pathway-fly)等在高温组和低氧组中均有显著富集。研究结果表明,miRNA在刺参响应高温、低氧胁迫中参与了一系列关键的免疫相关过程的调控。

In recent years,continuous high temperatures and rainstorm events in Shandong and Hebei Provinces,China,triggered by El Nino,have resulted in mass mortality of sea cucumbers,seriously restricting the sustainable development of sea cucumber aquaculture.In present study,the coelomic fluid cells of Apostichopus japonicus were chosen as experimental materials to construct the differentially expressed miRNA profile and conduct the functional analysis of their target genes under heat stress and hypoxia stresses.A total of 20 differentially expressed miRNAs were identified in heat stress group and 11 of them were up-regulated and 9 of them were down-regulated.A total of 10 differentially expressed miRNAs were identified in hypoxia group and 6 of them were up-regulated and 4 of them were down-regulated.And the common differentially expressed miRNAs both in heat stress and hypoxia groups were novel-23,novel-43,novel-45,novel-49,miR-200-3p and miR-71.The target genes of these differentially expressed miRNAs were predicted with miRanda software and 14827 target genes were predicted for 20 differentially expressed miRNAs in heat stress group,and 8577 target genes were predicted for 10 differentially expressed miRNAs in hypoxia group.KEGG enrichment analysis showed 36 significantly enriched pathways in heat stress group and 37 pathways significantly enriched in hypoxia group.PI3K-Akt signaling pathway,pathways in cancer,focal adhesion,MAPK signaling pathway-fly and others were significantly enriched both in heat stress and hypoxia group,suggesting that miRNA involve in the regulation of immune-related biological process in responding heat stress and hypoxia stresses in sea cucumber.