Differential expression of microRNAs in PK-15 cells infected with Pseudorabies virus-Fa strain

Abstract:

[Objective] The aim was to analyze the microRNAs expression in PK-15 cells infected with Pseudorabies virus-Fa. [Methods] Illumina deep sequencing was used to identify and analyze the differentially expressed miRNAs in PK-15 cells with or without PRV-Fa infection. The sequencing results for selected miRNAs were confirmed with RT-qPCR. The host target genes were predicted and Gene Ontology (GO) analysis was done. [Results] We identified 384 and 405 miRNAs in the infected and uninfected cells, respectively. Overall, 127 miRNAs were expressed significantly differently after infected with PRV-Fa (60 upregulated and 67 downregulated). RT-qPCR showed that the expressions of miRNAs were consistent with the sequencing results. GO analysis showed that miRNAs were widely involved in signal transduction, cells metabolism, immune response, and gene expression. And miR-10b, miR-16, miR-18a, miR-19b, miR-20a, miR-145-5p, miR-146a, miR-181a and miR-499-5p were associated with immune response. In the regulatory network of target genes, ssc-miR-30d and ssc-miR-30a-5p were in the key positions. Moreover, five miRNAs encoded by PRV-Fa were identified, PRV-miR-LLT2 and PRV-miR-LLT4 were targeted to PRV EPO gene. [Conclusion] The infection of PRV-Fa has significant influence on PK-15 cells encoding miRNAs.

Key words:Pseudorabies virus, microRNAs, Illumina deep sequencing, PK-15 cells