| 摘要: |
| 旨在比较分析捻转血矛线虫敏感虫株和耐药虫株miRNA表达谱，探讨miRNA与捻转血矛线虫丙硫咪唑耐药性相关基因之间的调控机制。运用Illumina Hiseq2000平台进行测序并进行cDNA文库的构建，利用RNAhybrid、Miranda和TargetScan对测序得到的数据进行靶基因预测，并取交集作为miRNA的靶基因预测结果，使用Bowtie、RepeatMasker、MIREAP、Rfam、miRBase和DAVID等生物信息学分析软件和权威数据库系统，筛选出差异的miRNA以及与捻转血矛线虫耐药相关的miRNA并进行靶基因预测，同时对KEGG pathway和GO功能富集到的靶基因和可能参与调控的通路进行预测分析。结果表明，敏感虫株和耐药虫株共筛选显著差异表达的miRNA共294个，其中113个上调，181个下调。对miRNA和其靶向调节的mRNA进行关联分析，共关联到1 770 个miRNA-mRNA对为负调控关系，其中涉及到274个差异的miRNA和603个差异的mRNA。显著差异表达的miRNA靶基因被注释到了1 430条GO terms和327条KEGG pathway，富集到FoxO信号通路(FoxO signaling pathway)，mTOR信号通路(mTOR signaling pathway)，PI3K-Akt信号通路(PI3K-Akt signaling pathway)等与耐药相关的一些抗性通路。综上，通过对miRNA表达谱分析以及miRNA-mRNA靶向分析，发现捻转血矛线虫敏感虫株和耐药虫株显著差异表达的miRNA和其对应的靶基因，并对部分耐药相关通路中存在的靶基因及显著差异的靶基因所在的通路进行筛选和分析，为后续进一步通过转录组学深层次挖掘捻转血矛线虫产生耐药性的关键基因及相关调控分子提供科学依据。 |
| 关键词: 捻转血矛线虫 丙硫咪唑 耐药性 转录组 miRNA |
| DOI：10.11841/j.issn.1007-4333.2021.06.10 |
| 投稿时间：2020-10-03 |
| 基金项目:国家自然科学基金项目(31760731)；内蒙古自治区科技计划项目(201702074) |
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| Comparative analysis of miRNA-mRNA association between albendazole-susceptible and-resistant strains of Haemonchus contortus |
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WANG Tengyu1,CHEN Xindi1,ZHAI Shuai1,SHI Yaqin1,LIU Chunxia2,WANG Wenlong1*
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| (1.Key Laboratory of Clinical Diagnosis and Treatment Technology in Animal Disease of Ministry of Agriculture/College ofVeterinary Medicine, Inner Mongolia Agricultural University, Hohhot 010018, China;2.College of Life Science, Inner Mongolia Agricultural University, Hohhot 010018, China) |
| Abstract: |
| The aim of this study was to compare miRNA expression profiles of sensitive and resistant strains of Haemonchus contortus, and to explore the regulatory mechanism between miRNA and albendazole resistance related genes of H. contortus. Illumina Hiseq2000 platform was used to sequence and construct the cDNA library. RNAhybrid, Miranda, TargetScan were used to predict the target gene of the sequenced data, and then their intersection was taken as the miRNA prediction result. Bioinformatics analysis softwares such as Bowtie, RepeatMasker, MIREAP, Rfam, miRbase, DAVID and authoritative database systems were used to screen differentially expressed miRNAs and miRNAs associated with drug resistance of H. contortus for target genes prediction. Target genes enriched by KEGG pathway and GO function and possible pathways involved in regulation were predicted and analyzed. The results showed that 294 miRNAs were differentially screened from sensitive and resistant strains, of which 113 were up-regulated and 181 were down-regulated. A total of 1 770 miRNA-mRNA pairs are negatively correlated with miRNA, including 274 differentially expressed miRNAs and 603 differentially expressed mRNAs. The target genes of the differentially expressed miRNA were annotated to 1 430 GO terms and 327 KEGG pathways, and enriched to FoxO signaling pathway, mTOR signaling pathway, PI3K-Akt signaling pathway and other resistance-related pathways. In summary, through the analysis of miRNA expression profile and miRNA-mRNA targeting analysis, it was found that there were significantly different miRNA and the corresponding target genes between the sensitive and resistant strains of H. contortus. The target genes in some drug-resistance related pathways and the pathways in which the significantly different target genes were located, screened and analyzed. This study provided references for the further exploration of the genes and regulatory molecules related to drug resistance of H. contortus. |
| Key words: Haemonchus contortus albendazole drug resistance transcriptome miRNA |
