猪圆环病毒Ⅱ型感染3D4/2细胞的转录组学分析

doi:10.16431/j.cnki.1671-7236.2024.01.005

Abstract

Abstract: 【Objective】 The aim of this study was to screen the differentially expressed genes of Porcine circovirus type Ⅱ (PCV2) infected porcine alveolar macrophage cell line (3D4/2) by transcriptome sequencing, and lay a foundation for understanding the host immune cell mechanism of PCV2 infection and the development of anti-PCV2 infection drugs.【Method】 PCV2 NJ2002 strain with multiplicity of infection (MOI) equal to 1 was used to treat 3D4/2 cells, and transcriptome sequencing was performed using Illumina NovaSeq 6000 sequencing platform.DeSeq 2.0 software was used for differential expressed genes analysis.GO and KEGG function analysis and transcription factor targeting analysis were also performed for the differentially expressed genes.The genes related to immunity and apoptosis were selected for Real-time quantitative PCR verification, and the protein expression levels of intracellular phosphatidylinositol kinase (PI3K), phosphorylated intracellular phosphatidylinositol kinase (p-PI3K), protein kinase B (Akt) and phosphorylated protein kinase B (p-Akt) were detected by Western blotting.【Result】 Transcriptome sequencing results showed that compared with the control group, 713 differentially expressed genes were obtained in PCV2 infection group, of which 313 were up-regulated and 400 were down-regulated. GO function and KEGG pathway enrichment analysis showed that differentially expressed genes were related to immune response, mainly concentrated in extracellular matrix receptor interaction pathway, metabolic pathway, HIF-1 signaling pathway, viral carcinogenesis, PI3K-Akt signaling pathway.The results of Real-time quantitative PCR were consistent with those of transcriptome sequencing.Western blotting results showed that the phosphorylation levels of PI3K and Akt were significantly increased after PCV2 infected 3D4/2 cells for 24 h (P<0.05).Transcription factor targeting analysis showed that differentially expressed genes were closely related to nuclear transcription factor Y subunit β (NFYB) and ETS transcription factor (ELK4).【Conclusion】 PCV2 might affect the downstream inflammatory signaling pathway and apoptosis signaling pathway to enhance its self-replication ability through the PI3K-Akt signaling pathway.NFYB and ELK4 transcription played important roles in PCV2 infection and could be considered as targets for anti-PCV2 drugs.The results provided theoretical basis for further understanding the mechanism of PCV2 infection in host immune cells and related drug development.

Key words: Porcine circovirus type Ⅱ; 3D4/2 cell; transcriptomics sequencing; differentially expressed genes; transcription factor

CLC Number:

S852.3

CHEN Hongling, ZHAO Yi, CHEN Jiaji, WEI Qiuxu, LI Ximeng, FENG Guoyue, HU Kunxiang, HU Tingjun. Transcriptomic Analysis of 3D4/2 Cells Infected with Porcine Circovirus Type Ⅱ[J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(1): 42-51.

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Transcriptomic Analysis of 3D4/2 Cells Infected with Porcine Circovirus Type Ⅱ

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