High-Throughput Sequencing of MicroRNAs in Adenovirus Type 3 Infected Human Laryngeal Epithelial Cells

Yuhua Qi,Yunfeng Shan,Hua Wang,Zuhong Lu,Lunbiao Cui,Yiyue Ge,Xiling Guo,Jing Tu,Jun Shan,Wenting Shi,Shuchun Li,Zhiyang Shi

doi:10.1155/2010/915980

Abstract

Adenovirus infection can cause various illnesses depending on the infecting serotype, such as gastroenteritis, conjunctivitis, cystitis, and rash illness, but the infection mechanism is still unknown. MicroRNAs (miRNA) have been reported to play essential roles in cell proliferation, cell differentiation, and pathogenesis of human diseases including viral infections. We analyzed the miRNA expression profiles from adenovirus type 3 (AD3) infected Human laryngeal epithelial (Hep2) cells using a SOLiD deep sequencing. 492 precursor miRNAs were identified in the AD3 infected Hep2 cells, and 540 precursor miRNAs were identified in the control. A total of 44 miRNAs demonstrated high expression and 36 miRNAs showed lower expression in the AD3 infected cells than control. The biogenesis of miRNAs has been analyzed, and some of the SOLiD results were confirmed by Quantitative PCR analysis. The present studies may provide a useful clue for the biological function research into AD3 infection.

Highlights

Adenovirus (AD) belongs to a family of nonenveloped icosahedral viruses containing a double-stranded DNA genome, Adenoviridae, and genus Mastadenovirus
We analyzed the miRNA expression profiles from adenovirus type 3 (AD3) infected Human laryngeal epithelial (Hep2) cells using a Sequencing by Oligonucleotide Ligation and Detection (SOLiD) deep sequencing. 492 precursor miRNAs were identified in the AD3 infected Hep2 cells, and 540 precursor miRNAs were identified in the control
Cytopathic effect (CPE) was first observed at 24 hours post infection and progressed to moderate and severe CPE at 72 hours and 96 hours, respectively. miRNA extracted from cells after 72 hours of infection was used for SOLiD deep sequencing

Summary

Introduction

Adenovirus (AD) belongs to a family of nonenveloped icosahedral viruses containing a double-stranded DNA genome, Adenoviridae, and genus Mastadenovirus. Studies showed that AD can evade host immune responses, such as inhibition of interferon functions by RNA and E1A, and inhibition of intrinsic cellular apoptosis in infected cells [2]. Several studies demonstrated that RNA interference (RNAi) can be used for treatment with viral infection in vitro, such as hepatitis B virus, coxsackievirus B3, and Coxsackievirus A16 [6,7,8]. These findings indicate that those specific siRNAs targeted viruses are not as effective as expected

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