A型塞内卡病毒感染PK-15细胞环状RNA表达谱分析

doi:10.16431/j.cnki.1671-7236.2023.01.028

摘要/Abstract

摘要： 【目的】分析A型塞内卡病毒（Seneca virus A，SVA）感染猪肾上皮细胞（PK-15）后环状RNA （circular RNA，circRNA）表达谱的变化，探究circRNAs在SVA感染过程中发挥的潜在调控作用。【方法】本研究基于Illumina HiSeq 2000平台对SVA感染及对照PK-15细胞circRNAs转录组进行测序，并对差异表达circRNAs的来源基因进行GO功能及KEGG通路富集分析，通过实时荧光定量PCR对新发现的circRNAs进行表达水平鉴定。【结果】转录组测序结果显示，SVA感染组、PK-15细胞对照组中共有432种circRNAs被发现。在SVA感染PK-15细胞中，87种circRNAs表达水平相对对照组PK-15细胞显著上调，74种circRNAs表达水平显著下调。对差异表达circRNAs的来源基因进行GO功能注释分析表明，SVA感染组差异表达circRNAs来源基因主要富集于核仁、细胞器膜和细胞大分子代谢、发育等生理过程。KEGG通路富集结果显示，SVA感染组差异表达circRNAs来源基因主要富集于胞吞过程、cAMP信号通路、Rap1信号通路、Ras信号通路。对6种新发现的显著差异表达的circRNAs进行实时荧光定量PCR验证，结果显示，其表达水平与高通量测序结果趋势一致。【结论】本研究首次对SVA感染PK-15细胞的circRNAs表达谱进行差异分析，发现差异表达的circRNAs广泛参与动物机体大分子代谢、胞吞及细胞增殖、黏附、抗病毒过程，为进一步探究circRNAs在SVA感染过程中的分子机制提供了理论依据。

关键词: 环状RNA (circRNA); A型塞内卡病毒(SVA); 高通量测序; PK-15细胞

Abstract: 【Objective】 The purpose of the test was to analyze the effect of Seneca virus A (SVA) infection on the expression profile of circular RNAs (circRNAs) in pig renal epithelial cells (PK-15), and to explore the potential regulatory role of circRNAs during SVA infection.【Method】 The circRNAs transcriptome of SVA-infected and control PK-15 cells were sequenced based on the Illumina HiSeq 2000 platform.The source genes of the differentially expressed circRNAs were subjected to GO function and KEGG pathway enrichment analysis, and the expression levels of the newly discovered circRNAs were identified by Real-time quantitative PCR.【Result】 Transcriptome sequencing results revealed that a total of 432 novel circRNAs were identified in SVA-infected and control PK-15 cells.In SVA-infected PK-15 cells, the expression levels of 87 circRNAs were significantly up-regulated and 74 circRNAs were significantly down-regulated compared to control PK-15 cells. GO functional analysis of the source genes of differentially expressed circRNAs indicated that the differentially expressed circRNAs source genes in SVA-infected PK-15 cells were mainly enriched in the nucleolus, organelle membranes, and physiological processes such as cellular macromolecular metabolic processes and development.The results of KEGG pathway enrichment analysis showed that the significantly differentially expressed circRNAs source genes in the SVA-infected PK-15 cells were mainly enriched in the cytokinesis process, cAMP signaling pathway, Rap1 signaling pathway and Ras signaling pathway.The results of Real-time quantitative PCR verification of the 6 significantly differentially expressed circRNAs demonstrated that their expression levels were consistent with the high-throughput sequencing.【Conclusion】 This was the first report of differential analysis of the circRNAs expression profiles of SVA-infected PK-15 cells.It was found that the differentially expressed circRNAs were widely involved in macromolecular metabolism, cytokinesis and cell proliferation, adhesion and antiviral processes, and this study provided a reference for the further exploration of the molecular mechanism of circRNAs during SVA-infection.

Key words: circular RNA (circRNA); Seneca virus A (SVA); high-throughput sequencing; PK-15 cell

中图分类号:

S852.65⁺9.6

陈彦希, 王晨, 罗愿, 周远成, 徐志文, 彭远义, 宋振辉, 刘骁. A型塞内卡病毒感染PK-15细胞环状RNA表达谱分析[J]. 中国畜牧兽医, 2023, 50(1): 280-289.

CHEN Yanxi, WANG Chen, LUO Yuan, ZHOU Yuancheng, XU Zhiwen, PENG Yuanyi, SONG Zhenhui, LIU Xiao. circRNA Expression Profile Analysis of PK-15 Cells Response to Seneca Virus A Infection[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(1): 280-289.

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