Zi Chen, Zeng Dexin, Zhou Jiyong, Dai Jianjun, Jiang Luyan, Xue Feng, Jiang Yuan, Li Baoguang
MOE Joint International Research Laboratory of Animal Health and Food safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, 210095, China.
APFIC, Jiangsu Entry-Exit Inspection and Quarantine Bureau, Nanjing, 210001, China.
Genes Genomics. 2018 Apr;40(4):361-371. doi: 10.1007/s13258-017-0635-3. Epub 2017 Dec 14.
MicroRNAs are implicated in many cellular processes such as cell differentiation and development, tumorigenesis, and immune regulation. In this study, miR192 was detected using quantitative real-time polymerase chain reaction (qRT-PCR) when MDBK cells were exposed to Escherichia coli. Cells with malfunction of bta-miR-192 were established using transcription activator-like effector nuclease (TALEN) technology. Finally, bta-miR-192 mutant cells were screened for differentially expressed genes using RNA-sequencing (RNA-seq). The results showed that miR192 significantly decreased in cells exposed to E. coli F18ac and E. coli K88ac. The RNA-seq results showed that 1673 differentially expressed transcripts were identified; 890 genes were upregulated and 775 genes were downregulated. With the gene ontology enrichment analysis, 431 differentially expressed genes (DEGs) were classified into 937 gene ontology terms. The pathway enrichment analysis showed that 535 genes were involved in 254 pathway terms. Interestingly, most of these DEGs were associated with the pathways in cancers or infectious diseases. When the selected DEGs (n = 162) in these pathways were intersected with 120 differential transcripts, 11 DEGs were identified. Subsequently, several genes associated with regulation, cancers, or viral infections, such as LEF1, AXIN2, MX1, and FCGR2B, were identified among the DEGs using functional analysis. Furthermore, associations between bta-miR-192 and DEGs were detected by intersecting the bta-miR-192's target genes with the DEGs, indicating that three genes including CBL, DICER1 and TRERF1 were involved in this relationship. These findings provided useful guidance for investigating the role played by bta-miR-192 in cellular functionality in bovine cells.
微小RNA参与许多细胞过程，如细胞分化与发育、肿瘤发生和免疫调节。在本研究中，当MDBK细胞暴露于大肠杆菌时，使用定量实时聚合酶链反应（qRT-PCR）检测miR192。利用转录激活样效应核酸酶（TALEN）技术构建bta-miR-192功能缺失的细胞。最后，使用RNA测序（RNA-seq）筛选bta-miR-192突变细胞中的差异表达基因。结果表明，暴露于大肠杆菌F18ac和大肠杆菌K88ac的细胞中miR192显著降低。RNA-seq结果显示，共鉴定出1673个差异表达转录本；890个基因上调，775个基因下调。通过基因本体富集分析，431个差异表达基因（DEG）被归类到937个基因本体术语中。通路富集分析表明，535个基因参与254个通路术语。有趣的是，这些DEG中的大多数与癌症或传染病通路相关。当这些通路中选定的DEG（n = 162）与120个差异转录本相交时，鉴定出11个DEG。随后，通过功能分析在DEG中鉴定出几个与调节、癌症或病毒感染相关的基因，如LEF1、AXIN2、MX1和FCGR2B。此外，通过将bta-miR-192的靶基因与DEG相交，检测到bta-miR-192与DEG之间的关联，表明包括CBL、DICER1和TRERF1在内的三个基因参与了这种关系。这些发现为研究bta-miR-192在牛细胞的细胞功能中所起的作用提供了有用的指导。
