Abstract
Transmissible gastroenteritis virus (TGEV; Coronaviridae family) causes huge economic losses to the swine industry. MicroRNAs (miRNAs) play a regulatory role in viral infection and may be involved in the mammalian immune response. Here, we report a comprehensive analysis of host miRNA expression in TGEV-infected swine testis (ST) cells. Deep sequencing generated 3,704,353 and 2,763,665 reads from uninfected ST cells and infected ST cells, respectively. The reads were aligned to known Sus scrofa pre-miRNAs in miRBase 19, identifying 284 annotated miRNAs. Certain miRNAs were differentially regulated during TGEV infection. 59 unique miRNAs displayed significant differentially expression between the normal and TGEV-infected ST cell samples: 15 miRNAs were significantly up-regulated and 44 were significantly down-regulated. Stem-loop RT-PCR was carried out to determine the expression levels of specific miRNAs in the two samples, and the results were consistent with those of sequencing. Gene ontology enrichment analysis of host target genes demonstrated that the differentially expressed miRNAs are involved in regulatory networks, including cellular process, metabolic process, immune system process. This is the first report of the identification of ST cell miRNAs and the comprehensive analysis of the miRNA regulatory mechanism during TGEV infection, which revealed the miRNA molecular regulatory mechanisms for the viral infection, expression of viral genes and the expression of immune-related genes. The results presented here will aid research on the prevention and treatment of viral diseases.
Highlights
Transmissible gastroenteritis virus (TGEV), belonging to the Coronaviridae family, is the pathogen of transmissible gastroenteritis of swine (TGE)
To investigate the miRNA expression profiles of infected swine testis (ST) cells, ST cells were infected with TGEV SC strain and the normal ST cell line was used as a control. 15–25 nt sRNAs were isolated and analyzed by deep sequencing
The analysis showed the target genes of the differential expressed miRNAs belong to the T cell receptor signaling pathway, which is involved in the regulation of T cell development, cytokine production and the activation of induced cell death, which is crucial for anti-virus activity
Summary
TGEV Sichuan strain and the ST cell line (Provided by Animal Biotechnology Center of Sichuan Agricultural University) were used in this study. Cell cultures were performed at 106 cells/ 175 cm dishes and were maintained under standard conditions of 37°C and 5% CO2 in modified RPMI-1640 nutrient solution (Thermo Fisher Scientific, Waltham, UK), supplemented with 10% heat-inactivated fetal bovine serum and 50 mg/ml penicillin/streptomycin antibiotic solution (GIBCO, Beijing, China). When the cells were 70% confluent in the culture dishes, they were infected with TGEV at 10 PFU per cell. The infected and control cells groups were harvested at 72 post infection (hpi), and resuspended in Trizol (Invitrogen, Shanghai, China) and stored at-70°C. All infected and control samples were combined into a single control group and a single infected group, respectively
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
