Parallel mRNA and MicroRNA Profiling of HEV71-Infected Human Neuroblastoma Cells Reveal the Up-Regulation of miR-1246 in Association with DLG3 Repression

Abstract

Human enterovirus 71 (HEV71) has emerged as the leading cause of viral encephalitis in children in most Asian countries. The roles of host miRNAs in the neurological pathogenesis of HEV71 infection remain unknown. In the present study, comprehensive miRNA expression profiling in HEV71-infected human neuroblastoma SH-SY5Y cells was performed using the Affymetrix Gene Chip microarray assay and was validated using real-time RT-PCR. Among the 69 differentially expressed miRNAs, miR-1246 was specifically induced by HEV71 infection in human neuroblastoma cells, but inhibition of miR-1246 failed to affect HEV71 replication. Parallel mRNA and microRNA profiling based on the 35 K Human Genome Array identified 182 differentially regulated genes. Target prediction of miR-1246 and network modeling revealed 14 potential target genes involved in cell death and cell signaling. Finally, a combined analysis of the results from mRNA profiling and miR-1246 target predication led to the identification of disc-large homolog 3 (DLG3), which is associated with neurological disorders, for further validation. Sequence alignment and luciferase reporter assay showed that miR-1246 directly bound with the 3′-UTR of DLG3 gene. Down-regulation of miR-1246 induced significant changes in DLG3 expression levels in HEV71-infected SHSY5Y cells. Together, these results suggested that miR-1246 might play a role in neurological pathogenesis of HEV71 by regulating DLG3 gene in infected cells. These findings provide new information on the miRNA and mRNA profiles of HEV71-infected neuroblastoma cells. The biological significance of miR-1246 and DLG3 during the course of HEV71 infection deserves further investigation.

Systematic Identification and Bioinformatic Analysis of MicroRNAs in Response to Infections of Coxsackievirus A16 and Enterovirus 71

“…One of the 27 differentially expressed miRNAs identified in our study, miR-1246, was increased by 3.95-fold at the expression level in EV71-infected cells. This finding was consistent with a previous study by Xu and colleagues [30]. They also found the EV71-mediated upregulation of miR-1246 in infected cells and further unveiled that miR-1246 suppressed the expression of disc-large homolog 3 (DLG3) gene by directly binding the 3′-UTR sequence, which contributed to the neurological disorders induced by EV71 infection.…”

“…The EV71-mediated miRNA expression profile identified in present study displayed a little difference from that performed in previous report by Xu et al [30]. This discrepancy might be largely due to different cell types used in viral infection between studies.…”

Octamer 4/microRNA‐1246 signaling axis drives Wnt/β‐catenin activation in liver cancer stem cells

“…miR‐1246 promotes proliferation, migration and invasion in human cervical carcinoma cells though targeting thrombospondin 2 . Up‐regulation of miR‐1246 in human neuroblastoma cells is associated with discs large homolog 3 . miR‐1246 have also been found to promote angiogenesis by activating small mothers of decapentaplegic 1/5/8 signaling elicited by promyelocytic leukemia down‐regulation in endothelial cells .…”

Upregulation of miR-181 Decreases c-Fos and SIRT-1 in the Hippocampus of 3xTg-AD Mice

“…F) However, c-Fos mRNA levels were similar between genotypes at 12 months in the ventral hippocampus. G) Representative blots of the data presented in A-B, D-E. * p < 0.05. diseases [44,45]. Fourty-eight hours after transfection of miR-181 or a non-related sequence as a negative control, the steady-state levels of SIRT-1 and c-Fos were determined.…”