MicroRNA expression profile of human umbilical vein endothelial cells in response to Coxsackievirus A10 infection

Coxsackievirus A10 (CV-A10) constitutes one of the major pathogens of hand, foot, and mouth disease (HFMD), which can cause mild to severe illness and even death. Most of these severe and death cases were closely associated with their neurological impairments, but the underlying mechanism of neuropathological injury induced by CV-A10 infection has not been elucidated. MicroRNAs (miRNAs), implicated in the regulation of gene expression in a posttranscriptional manner, play a vital role in the pathogenesis of various central nervous systems (CNS) diseases; thereby they are served as diagnostic biomarkers and are emerging as novel therapeutic targets for CNS injuries. To gain insights in the CV-A10-induced regulation of host miRNA-processing machinery, we employed high-throughput sequencing to identify differentially expressed miRNAs in CV-A10-infected HUVEC cells and further analyzed the potential functions of these miRNAs during CV-A10 infection. The results showed that CV-A10 infection could elicit 189 and 302 significantly differentially expressed miRNAs in HUVEC cells at 24 hpi and 72 hpi, respectively, as compared with the uninfected control. For virus infection, single-cell suspension at a density of 5 × 105 cells/ml was added into 6-well plates. When the cells were reached 80% confluences, the cells were incubated with CV-A10 (sub-genotype C, GenBank: MN557275), which isolated from an epidemic in Xiangyang, China, in 2017, at a multiplicity of infection (MOI) of 1 at 37°C for 2 h absorption. Subsequently, the infected-cells were continued to culture in RPMI-1640 containing 2% FBS and antibiotics at 37 °C in an atmosphere of 5% CO2, and collected with a cell scraper at 0, 24 and 72 h. Cells infected with CV-A10 at 0 hpi were regarded as control.