Limits...
Differential expression of miRNAs in enterovirus 71-infected cells.

Xun M, Ma CF, Du QL, Ji YH, Xu JR - Virol. J. (2015)

Bottom Line: Increasing evidence suggests that miRNAs act as key effector molecules in the complicated pathogen-host interactions.Then, the results of microarray were further confirmed by quantitative RT-PCR.The expression levels of 8 most differentially up-regulated miRNAs and 3 most differentially down-regulated miRNAs were confirmed by qRT-PCR.

View Article: PubMed Central - PubMed

Affiliation: Department of Immunology and Microbiology, Medical School of Xi'an Jiaotong University, Xi'an, 710061, Shaanxi, China. xunmeng@mail.xjtu.edu.cn.

ABSTRACT

Background: 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.

Show MeSH

Related in: MedlinePlus

The morphological changes of EV71-induced cytopathic effect in RD cells were observed under a light microscope at 20× magnification at 48 h p.i.. RD cells were infected with EV71 at a m.o.i. of 0.1. (A) EV71-infected RD cells exhibited severe CPE appearance in response to virus replication at 48 h p.i..; (B) Uninfected RD cells showed normal morphology.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
getmorefigures.php?uid=PMC4416288&req=5

Fig1: The morphological changes of EV71-induced cytopathic effect in RD cells were observed under a light microscope at 20× magnification at 48 h p.i.. RD cells were infected with EV71 at a m.o.i. of 0.1. (A) EV71-infected RD cells exhibited severe CPE appearance in response to virus replication at 48 h p.i..; (B) Uninfected RD cells showed normal morphology.

Mentions: RD cells which are frequently used to isolate EV71 from clinical specimens are highly susceptible to EV71. After 48 h of infection, EV71 induced a severe CPE in RD cells (Figure 1).Figure 1


Differential expression of miRNAs in enterovirus 71-infected cells.

Xun M, Ma CF, Du QL, Ji YH, Xu JR - Virol. J. (2015)

The morphological changes of EV71-induced cytopathic effect in RD cells were observed under a light microscope at 20× magnification at 48 h p.i.. RD cells were infected with EV71 at a m.o.i. of 0.1. (A) EV71-infected RD cells exhibited severe CPE appearance in response to virus replication at 48 h p.i..; (B) Uninfected RD cells showed normal morphology.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4416288&req=5

Fig1: The morphological changes of EV71-induced cytopathic effect in RD cells were observed under a light microscope at 20× magnification at 48 h p.i.. RD cells were infected with EV71 at a m.o.i. of 0.1. (A) EV71-infected RD cells exhibited severe CPE appearance in response to virus replication at 48 h p.i..; (B) Uninfected RD cells showed normal morphology.
Mentions: RD cells which are frequently used to isolate EV71 from clinical specimens are highly susceptible to EV71. After 48 h of infection, EV71 induced a severe CPE in RD cells (Figure 1).Figure 1

Bottom Line: Increasing evidence suggests that miRNAs act as key effector molecules in the complicated pathogen-host interactions.Then, the results of microarray were further confirmed by quantitative RT-PCR.The expression levels of 8 most differentially up-regulated miRNAs and 3 most differentially down-regulated miRNAs were confirmed by qRT-PCR.

View Article: PubMed Central - PubMed

Affiliation: Department of Immunology and Microbiology, Medical School of Xi'an Jiaotong University, Xi'an, 710061, Shaanxi, China. xunmeng@mail.xjtu.edu.cn.

ABSTRACT

Background: 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.

Show MeSH
Related in: MedlinePlus