捻转血矛线虫丙硫咪唑耐药相关lncRNA的转录组学测序与分析

Transcriptome sequencing and analysis of lncRNA related to albendazole resistant strain of Haemonchus contortus

摘要:

分析捻转血矛线虫敏感株和耐药株中长链非编码RNA (Long non-coding RNA，lncRNA)的表达谱，探讨lncRNA与捻转血矛线虫丙硫咪唑耐药机制的关联性，为捻转血矛线虫耐药机理提供依据。文中对捻转血矛线虫敏感株和耐药株进行cDNA测序文库构建，使用Illumina HiSeq 4000平台进行双端测序，筛选出差异的lncRNA，基于顺式调控 (cis) 和反式调控 (trans) 对差异显著的lncRNA进行靶基因预测，并对靶基因进行过Gene Ontology (GO)功能注释和KEGG Pathway富集分析，利用FPKM法估计lncRNA和mRNA的表达水平。结果表明，敏感株和耐药株候选lncRNA分别为6 377和6 356个，两个文库中筛选出168个差异显著的lncRNA，其中在敏感株中表达上调有92个，表达下调76个。筛选得到的差异显著lncRNA候选靶基因416个，这些基因共注释到641条GO terms和92条信号通路；其中富集到耐药性相关的通路有药物代谢-其他酶、药物代谢-细胞色素P450、细胞色素P450对异生素的代谢等。综上表明，部分lncRNA介导的靶基因与捻转血矛线虫丙硫咪唑耐药性相关，lncRNA在捻转血矛线虫耐药性中具有潜在的重要的调节作用。探究了对于敏感虫株和耐药虫株中lncRNA的表达谱，发现了敏感虫株和耐药虫株中差异表达的lncRNA，有助于找出捻转血矛线虫如何抵抗丙硫咪唑的发生机制，为探讨捻转血矛线虫丙硫咪唑耐药机制提供科学的依据。

Abstract:

The objective of this study was to explore the link between long non-coding RNAs (lncRNAs) and resistance mechanism of albendazole in Haemonchus contortus by analyzing the gene expression profile of lncRNA in sensitive and resistant strains. This study provided the base for the resistance mechanism of Haemonchus contortus. In this experiment, the cDNA sequencing libraries were collected constructed for the sensitive and resistant strains of Haemonchus contortus, and the sequencing was performed using Illumina HiSeq 4000 platform, and differentially expressed lncRNAs (DEIncRNAs) were screened. Then the cis-targeted and trans-targeted genes of DElncRNAs were predicted, and the GO enrichment, KEGG pathway enrichment analysis were also made. The results displayed that 6 377 and 6 356 candidate lncRNA transcripts of sensitive and resistive strains were respectively obtained, 168 DElncRNAs of which were selected. Compared with resistive strains, there were 92 up-regulated and 76 down-regulated lncRNAs in sensitive strains. Meanwhile, 416 candidate target genes of DElncRNAs were obtained. The analytical results indicated that these genes participated in 641 GO terms and 92 signal pathways. The pathways involved in drug resistance are drug metabolism-other enzymes, drug metabolism-cytochrome P450, metabolism of xenobiotics by cytochrome P450 and so on. In conclusion, these findings inferred that some lncRNA-mediated target genes were associated with the resistance of Haemonchus contortus, and lncRNA may play an important role in the resistance of Haemonchus contortus. This study explored the expression profile of lncRNA in the Haemonchus contortus of sensitive and resistant strains. It was found that DElncRNAs in the sensitive and resistant strains, which helped to find out the Haemonchus contortus resistance mechanism of albendazole and provided a scientific basis for exploring the mechanism of resistance to Haemonchus contortus.