Identification and Characterization of MicroRNAs in Snakehead Fish Cell Line upon Snakehead Fish Vesiculovirus Infection.
Identification and Characterization of MicroRNAs in Snakehead Fish Cell Line upon Snakehead Fish Vesiculovirus Infection.
复制标题
黑鱼水泡病毒感染后黑鱼细胞系中 MicroRNA 的鉴定和表征。
DOI：
10.3390/ijms17020154
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发表时间：
2016-01-26
影响因子：
5.6
通讯作者：
Lin L
中科院分区：
文献类型：
--
作者：
Liu X;Tu J;Yuan J;Liu X;Zhao L;Dawar FU;Khattak MN;Hegazy AM;Chen N;Vakharia VN;Lin L
微小RNA（microRNAs，miRNAs）在真核生物的多种生物学过程中起着重要的调节作用，并被越来越多的研究者用来评估它们在病毒感染后细胞变化中的作用。乌鳢水泡病毒（SHVV）在过去的几年里已经引起了乌鳢的大量死亡。为了鉴定参与SHVV感染的特异性miRNA，我们对有或没有SHVV感染的黑鱼细胞系（SSN-1）进行了microRNA深度测序。通过与已知的斑马鱼miRNAs比对，共鉴定出205种已知的miRNAs，并利用MiRDeep 2软件鉴定出9种新的miRNAs。在205种已知的miRNAs中，分别有18种和143种在感染后3小时和24小时（POI）差异表达。从差异表达的miRNAs中随机选取5个，采用定量逆转录聚合酶链反应（qRT-PCR）验证其表达谱，其表达谱与microRNA测序结果一致。此外，对差异表达的宿主miRNA进行SHVV基因组的靶基因预测，并且预测总共10个和58个差异表达的miRNA分别在3小时和24小时结合SHVV基因组。通过qRT-PCR和Western印迹，使用其模拟物和抑制剂评估三种选定的miRNA（miR-130- 5 p、miR-214和miR-216 b）对SHVV增殖的影响。结果表明，三种miRNAs均能抑制SHVV的增殖，但具体的miRNAs抑制SHVV增殖的机制有待进一步研究。
MicroRNAs (miRNAs) play important roles in mediating multiple biological processes in eukaryotes and are being increasingly studied to evaluate their roles associated with cellular changes following viral infection. Snakehead fish Vesiculovirus (SHVV) has caused mass mortality in snakehead fish during the past few years. To identify specific miRNAs involved in SHVV infection, we performed microRNA deep sequencing on a snakehead fish cell line (SSN-1) with or without SHVV infection. A total of 205 known miRNAs were identified when they were aligned with the known zebrafish miRNAs, and nine novel miRNAs were identified using MiRDeep2 software. Eighteen and 143 of the 205 known miRNAs were differentially expressed at three and 24 h post-infection (poi), respectively. From the differentially-expressed miRNAs, five were randomly selected to validate their expression profiles using quantitative reverse transcription polymerase chain reaction (qRT-PCR), and their expression profiles were consistent with the microRNA sequencing results. In addition, the target gene prediction of the SHVV genome was performed for the differentially-expressed host miRNAs, and a total of 10 and 58 differentially-expressed miRNAs were predicted to bind to the SHVV genome at three and 24 h poi, respectively. The effects of three selected miRNAs (miR-130-5p, miR-214 and miR-216b) on SHVV multiplication were evaluated using their mimics and inhibitors via qRT-PCR and Western blotting. The results showed that all three miRNAs were able to inhibit the multiplication of SHVV; whereas the mechanisms underlying the SHVV multiplication inhibited by the specific miRNAs need to be further characterized in the future.