ETEC感染猪小肠上皮细胞初期差异表达cricRNA的筛选

doi:10.16431/j.cnki.1671-7236.2023.09.005

Abstract

Abstract: 【Objective】 The purpose of this experiment was to analyze the changes of circularRNA (circRNA) expression profile in host cells of porcine small intestinal epithelial cells (IPEC-J2) at the beginning of enterotoxigenic Escherichia coli (ETEC) infection,and explore the regulation mechanism of circRNA.【Method】 Transcriptome sequencing was performed on the ETEC infected IPEC-J2 cells using Illumina PE150 sequencing platform,and differentially expressed circRNAs were screened using bioinformatics online software.After GO function and KEGG pathway enrichment analysis,circRNAs associated with ETEC infection were screened.Coding potential of circRNAs was predicted by IRESfinder and PFAM 33.1 software.Five differentially expressed circRNAs were randomly selected,and the expression of circRNAs and the digestion of RNase R were verified by Real-time quantitative PCR.【Result】Transcriptome sequencing results showed that 201 differentially expressed circRNAs were obtained,of which 95 were up-regulated and 106 were down-regulated.GO functional enrichment analysis showed that circRNA-derived genes at the early stage of ETEC infection mainly affected cell morphology,cell metabolism and ubiquitin-protein ligase activit.KEGG pathway enrichment analysis indicated that circRNAs might be involved in amino acid metabolism pathway,ubiquitin-mediated proteolysis pathway,adhesion junction and actin cytoskeleton regulation.Four circRNAs with coding potential were identified,among which novel_circ_0009996 and source gene TNFAIP3 were co-enriched in NF-κB and TNF signaling pathways.Real-time quantitative PCR results showed that the expression of the 5 circRNAs were consistent with the sequencing results,and the differentially expressed circRNAs were all circular,and the sequencing results were accurate and reliable.【Conclusion】 circRNAs and the source genes were mainly involved in inflammatory response,post-transcriptional regulation of gene expression,cellular immunity,cytoskeleton,cell proliferation,apoptosis and other related biological processes.novel_circ_0009996 had coding potential,which was mainly concentrated in the NF-κB and TNF signaling pathways.The results provided a theoretical basis for further exploring the regulatory mechanism of circRNA in the early stage ETEC infection.

Key words: enterotoxigenic Escherichia coli(ETEC); porcine small intestinal epithelial cells; circRNA; infection

CLC Number:

S814.8

ZHU Kaiqing, ZHU Yue, LIU Yingying, CUI Yanan, JIANG Bingyu, LI Yan. Screening of Differentially Expressed cricRNA of Porcine Small Intestinal Epithelial Cells at Early ETEC infection[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(9): 3490-3498.

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Screening of Differentially Expressed cricRNA of Porcine Small Intestinal Epithelial Cells at Early ETEC infection

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