Salmonella enteritidis Hcp distribute in the cytoplasm and regulate TNF signaling pathway in BHK-21 cells
Salmonella enteritidis Hcp distribute in the cytoplasm and regulate TNF signaling pathway in BHK-21 cells
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肠炎沙门氏菌Hcp分布于BHK-21细胞胞质并调节TNF信号通路
DOI：
10.1007/s13205-020-02296-0
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发表时间：
2020-06
期刊：
影响因子：
2.8
通讯作者：
Shuai Lin
中科院分区：
文献类型：
--
作者：
Liming Zheng;Shenghua Wang;Mengyu Ling;Zhengmei Lv;Shuai Lin
肠炎沙门氏菌的溶血素协同调节蛋白（Hcp）已知是 T6SS 的结构蛋白和效应蛋白，但人们对 Hcp 在宿主细胞中的作用知之甚少。本研究通过质粒pEGFP-N1-hcp在BHK-21细胞中表达Hcp，结果表明Hcp的亚细胞定位主要在BHK-21细胞的细胞质中。转染质粒pCI-neo-hcp在BHK-21细胞中过表达Hcp，并进行mRNA测序分析差异表达基因，结果显示307个mRNA的表达水平发生变化（变化倍数>2，p<0.01）。其中，125个mRNA上调，182个mRNA下调。京都基因与基因组百科全书（KEGG）富集分析显示，差异表达基因在肿瘤坏死因子（TNF）信号通路、IL-17信号通路、细胞因子-细胞因子受体相互作用等方面富集。随后，我们选择了TNF信号通路的差异表达基因，并通过实时PCR验证了变化。结果与测序结果观察到的趋势一致。总之，我们证明肠炎沙门氏菌的Hcp引起BHK-21细胞胞质中TNF信号通路mRNA表达的变化。
Hemolysin-coregulated protein (Hcp) ofSalmonella enteritidisis known to be a structural and effector protein of the T6SS, but little is known about the role of Hcp in host cells. In this study, Hcp was expressed by plasmid pEGFP-N1-hcp in BHK-21 cells and the results showed that the subcellular localization of Hcp was predominantly in the cytoplasm of BHK-21 cells. When Hcp was over-expressed by transfecting plasmid pCI-neo-hcp in BHK-21 cells and mRNA sequencing was performed to analyze differentially expressed genes, the results showed a change in the expression levels of 307 mRNAs (fold change > 2, andp< 0.01). Amongst these, 125 mRNAs were up-regulated and 182 mRNAs were down-regulated. Kyoto encyclopedia of genes and genomes (KEGG) enrichment analysis showed that differentially expressed genes were enriched in tumor necrosis factor (TNF) signaling pathway, IL-17 signaling pathway, and cytokine-cytokine receptor interaction. Subsequently, we selected differentially expressed genes of TNF signaling pathway and verified the changes by real-time PCR. The results were consistent with the trend observed for the sequencing results. In conclusion, we demonstrated that Hcp ofSalmonella enteritidiscaused the change of mRNAs expression of TNF signaling pathway in the cytoplasm of BHK-21 cells.
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