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

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。作者用感染复数(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1782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验证实验结果显示,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。

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 resulTSshowed that the expression of 1782 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 resulTSshowed 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.