Enterovirus 71 (EV71) is one of the major etiological pathogens of
hand, foot and mouth disease (HFMD) and can cause severe cerebral and pulmonary complications and even fatality.
MicroRNAs (
miRNAs), a class of
small non-coding RNA molecules, play an important role in post-transcriptional regulation of gene expression and thereby influencing various physiological and
pathological processes. Increasing evidence suggests that
miRNAs act as key effector molecules in the complicated pathogen-host interactions. However, the roles of
miRNAs in EV71
infection and pathogenesis are not well understood.
METHODS: To identify special
miRNAs involved in EV71
infection, a microarray assay was performed to study the expression pattern of
miRNAs in EV71-infected human
rhabdomyosarcoma cells (RD cells) and uninfected RD cells. We further predicted the putative target genes for the dysregulated
miRNAs using the online bioinformatic algorithms (TargetScan, miRanda and PicTar) and carried out functional annotation including GO enrichment and KEGG pathway analysis for
miRNA predicted targets. Then, the results of microarray were further confirmed by quantitative RT-PCR.
RESULTS: Totally, 45 differentially expressed
miRNAs ware identified by microarray, among which 36
miRNAs were up-regulated and 9 were down-regulated. 7166 predicted target genes for the dysregulated
miRNAs were revealed by using TargetScan in conjunction with miRanda and PicTar. The GO annotation suggested that predicted targets of
miRNAs were enriched into the category of signal transduction, regulation of transcription, metabolic process,
protein phosphorylation, apoptotic process and immune response. KEGG pathway analysis suggested that these predicted target genes were involved in many important pathways, mainly including endocytosis and focal adhesion, MAPK signaling pathway,
hypertrophic cardiomyopathy, melanogenesis and ErbB signaling pathway. The expression levels of 8 most differentially up-regulated
miRNAs and 3 most differentially down-regulated
miRNAs were confirmed by qRT-PCR. The expressions of hsa-miR-4530, hsa-miR-4492, hsa-miR-6125, hsa-miR-494-3p,
hsa-miR-638, hsa-miR-6743-5p, hsa-miR-4459 and
hsa-miR-4443 detected by qRT-PCR were consistent with the microarray data.
CONCLUSION: These results might extend our understanding to the regulatory mechanism of
miRNAs underlying the pathogenesis of EV71
infection, thus strengthening the preventative and therapeutic strategies of HFMD caused by EV71.