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利用大肠杆菌和金黄色葡萄球菌刺激牛乳腺上皮细胞的微阵列数据分析鉴定几个关键基因。

Identification of several key genes by microarray data analysis of bovine mammary gland epithelial cells challenged with Escherichia coli and Staphylococcus aureus.

机构信息

Harbin Specialty Research Institute, Heilongjiang Academy of Land Reclamation Sciences, No. 101 Xiangfu Road, Xiangfang District, Harbin 150038, China; Research and Development Center, Heilongjiang Puhui Specialty Co., LTD., Harbin 150038, China.

出版信息

Gene. 2019 Jan 30;683:123-132. doi: 10.1016/j.gene.2018.10.004. Epub 2018 Oct 3.

Abstract

PURPOSE

This study was aimed at exploring the mechanisms and identifying the key candidate genes associated with S. aureus and E. coli mastitis.

METHODS

A public microarray dataset GSE24560 was downloaded. Differentially expressed genes (DEGs) were screened in E. coli- and S. aureus-infected primary bovine mammary gland epithelial cell (pBMEC) samples, and compared with control samples at 1 h, 6 h, and 24 h. A functional enrichment analysis was performed, and construction of a gene co-expression network was performed based on genes that showed consistent changes over time, which were identified using time series expression analysis. Then, a miRNA/TF regulatory network was constructed based on the genes in the co-expression network. The genes in the miRNA/TF regulatory network were screened for involvement in related diseases. Furthermore, the expression of several selected DEGs was further validated using real-time RT-PCR.

RESULTS

In total, 92 and 81 DEGs showed continuous differential expression over time in the E. coli- and S. aureus-inoculated groups. DEGs in the E. coli-inoculated group were associated with the RIG-I-like receptor signaling pathway, and those in the S. aureus-inoculated group were associated with the lysosome pathway. Time series expression analysis identified two gene clusters. NFKBIZ and GRO1 in the gene co-expression network were associated with inflammatory and defense responses. Moreover, several genes such as CXADR, APP, and CXCL2 in the miRNA/TF regulatory network, were associated with infection, inflammation, or stress-related diseases.

CONCLUSION

RIG-I like receptor pathway and several DEGs such as NFKBIZ, GRO1, CXCL2, and CXADR may play critical roles in the response to infection in pBMECs.

摘要

目的

本研究旨在探讨金黄色葡萄球菌和大肠杆菌乳腺炎相关的机制,并确定关键候选基因。

方法

下载公共微阵列数据集 GSE24560。在金黄色葡萄球菌和大肠杆菌感染原代牛乳腺上皮细胞(pBMEC)样本中筛选差异表达基因(DEGs),并与对照组样本在 1h、6h 和 24h 进行比较。进行功能富集分析,并基于随时间一致变化的基因构建基因共表达网络,使用时间序列表达分析进行构建。然后,基于共表达网络中的基因构建 miRNA/TF 调控网络。筛选 miRNA/TF 调控网络中的基因是否参与相关疾病。进一步使用实时 RT-PCR 验证选定的 DEGs 的表达。

结果

在大肠杆菌和金黄色葡萄球菌感染组中,共有 92 个和 81 个 DEG 表现出持续的时间依赖性差异表达。大肠杆菌感染组中的 DEGs 与 RIG-I 样受体信号通路相关,金黄色葡萄球菌感染组中的 DEGs 与溶酶体途径相关。时间序列表达分析确定了两个基因簇。基因共表达网络中的 NFKBIZ 和 GRO1 与炎症和防御反应有关。此外,miRNA/TF 调控网络中的几个基因,如 CXADR、APP 和 CXCL2,与感染、炎症或应激相关疾病有关。

结论

RIG-I 样受体途径和几个 DEGs,如 NFKBIZ、GRO1、CXCL2 和 CXADR,可能在 pBMEC 对感染的反应中发挥关键作用。

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