单增李斯特菌感染RAW264.7细胞转录组差异表达分析

Comparative Analysis of Transcriptome Differential Expression in RAW264.7 Cells Infected by Listeria monocytogenes

摘要:

为研究单核细胞增生李斯特菌（Listeria monocytogenes，LM）感染前后RAW264.7细胞基因表达谱变化，筛选关键功能基因和通路，探究其致病机制。作者以感染复数（MOI）10的LM 感染RAW264.7细胞6 h，提取细胞RNA进行转录组测序，选取感染组与对照组相比差异倍数≥1.5且P<0.05的差异表达基因进行GO和KEGG富集分析，并通过RT-qPCR验证部分差异基因。转录组结果显示，RAW264.7细胞在感染LM前后有1 782 个基因差异表达，其中上调为989，下调为793。GO分析结果主要涉及免疫系统过程、免疫反应、细胞程序性死亡调控等。KEGG分析结果显示，显著富集的通路有细胞因子-受体相互作用、肿瘤坏死因子信号通路、IL-17信号通路等。RT-qPCR验证试验结果显示，随机选择的11 个显著差异基因表达倍数趋势与测序结果一致。该研究为进一步深入研究LM的致病机制奠定了基础。

Abstract:

The alterations of gene expression profile of RAW 264.7 cells before and after infection by Listeria monocytogenes (LM) were explored, and the key functional genes and pathways were screened to lay a foundation for investigating the pathogenic mechanism. The RAW264.7 cells were treated with LM at a multiplicity of infection (MOI) of 10 for 6 h, and RNA was extracted for transcriptome sequencing. Differentially expressed genes (DEGs) in the infection group compared to the control group with a 1.5 fold change and a P<0.05 were screened and analyzed by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. Some DEGs were validated by RT-qPCR. The transcriptome results showed that the expression of 1 782 genes in RAW264.7 cells changed significantly before and after infection with LM, including 989 up-regulated and 793 down-regulated genes. GO enrichment analysis mainly involved the immune system process, immune response, and regulation of programmed cell death and so on. KEGG analysis showed that the significantly enriched pathways included cytokine-receptor interaction, tumor necrosis factor, and IL-17 signaling pathway and so on. The RT-PCR verification experimental results showed that the trend of fold change of expression in the randomly selected 11 significantly DESs was consistent with the sequencing results. This study lays a foundation for further in-depth exploration of the pathogenic mechanism of LM.