Zhang Qi-Lin, Zhu Qian-Hua, Zhang Feng, Xu Bin, Wang Xiu-Qiang, Chen Jun-Yuan
State Key Laboratory of Pharmaceutical Biotechnology, School of Life Science, Nanjing University, Nanjing, China.
LPS, Nanjing Institute of Geology and Paleontology, Nanjing, China.
Oncotarget. 2017 Aug 28;8(43):73590-73602. doi: 10.18632/oncotarget.20570. eCollection 2017 Sep 26.
Amphioxus is a key experimental animal for studying the evolution of vertebrate immune system. However, we still do not know about the roles of microRNAs (miRNAs) under viral stress in amphioxus. In this study, we sequenced six small RNA libraries (three biological replicates were included in the treatments challenged by the viral mimic, poly (I:C) (pIC) and control groups, respectively) from . A total of 151 known miRNAs, 197 new miRNAs (named novel_mir, including nine conserved miRNAs) were identified by deep sequencing from the six libraries. We primarily focused on differentially expressed miRNAs (DEMs) after pIC challenge. Next, we screened a total of 77 DEMs, including 27 down- and 50 up-regulated DEMs in response to pIC challenge. Furthermore, we used real-time quantitative PCR (qRT-PCR) to verify the expression levels of 10 randomly selected DEMs. Target genes likely regulated by DEMs were predicted, and functional enrichment analyses of these targets were performed using bioinformatics approach. MiRNA targets of DEMs are primarily involved in immune response, diseases, cancer and regulation process, and could be largely linked to 14 immune-related signaling pathways, including NF-kappa B, NOD-like receptor, RIG-I-like receptor and endocytosis. The present study for the first time explores key regulatory roles of miRNAs in the innate antiviral immune response in amphioxus, and will provide insight into the molecular basis of antiviral immunity and evolution of immune-related miRNAs.
文昌鱼是研究脊椎动物免疫系统进化的关键实验动物。然而，我们仍然不清楚微小RNA（miRNA）在文昌鱼病毒应激下所起的作用。在本研究中，我们对来自……的六个小RNA文库进行了测序（分别在病毒模拟物聚肌苷酸-聚胞苷酸（pIC）刺激处理组和对照组中各包含三个生物学重复）。通过对这六个文库进行深度测序，共鉴定出151个已知miRNA、197个新miRNA（命名为novel_mir，其中包括9个保守miRNA）。我们主要关注pIC刺激后差异表达的miRNA（DEM）。接下来，我们总共筛选出77个DEM，其中包括27个下调和50个上调的DEM，这些DEM是对pIC刺激的响应。此外，我们使用实时定量PCR（qRT-PCR）来验证随机选择的10个DEM的表达水平。预测了可能受DEM调控的靶基因，并使用生物信息学方法对这些靶基因进行了功能富集分析。DEM的miRNA靶标主要参与免疫反应、疾病、癌症和调控过程，并且在很大程度上可能与14条免疫相关信号通路有关，包括核因子κB、NOD样受体、RIG-I样受体和内吞作用。本研究首次探索了miRNA在文昌鱼先天性抗病毒免疫反应中的关键调控作用，并将为抗病毒免疫的分子基础以及免疫相关miRNA的进化提供见解。
