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Open Access Highly Accessed Research article

Differential apoptosis gene expressions of rhabdomyosarcoma cells in response to enterovirus 71 infection

Weifeng Shi1*, Xiang Li1, Xueling Hou1, Hongjun Peng1, Qingbo Jiang1, Mei Shi1, Yun Ji1, Xiping Liu1 and Jinbo Liu2

Author Affiliations

1 Department of Clinical Laboratory, The Third Affiliated Hospital of Suzhou University, Changzhou, Jiangsu, 213003, China

2 Department of Orthopedics, The Third Affiliated Hospital of Suzhou University, Changzhou, Jiangsu, 213003, China

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BMC Infectious Diseases 2012, 12:327  doi:10.1186/1471-2334-12-327

Published: 28 November 2012

Abstract

Background

Enterovirus 71 (EV71) infection can induce the apoptosis of infected cells. The aim of this study is to explore the effect of EV71 infection on apoptosis mechanisms in virus-infected human rhabdomyosarcoma (RD) cells.

Methods

The apoptosis of RD cells was examined using annexin V-FITC/PI by flow cytometry and cytokines were detected by ELISA. Cellular RNA was extracted and transcribed to cDNA. PCR array was employed to analyze the expressions of 84 apoptotic genes from EV71-infected RD cells at 8 and 20 h postinfection, respectively. In addition, the expressions of FasL, caspase, AKT2, JNK1/2, c-Jun and NF-κB proteins were detected by western blotting.

Results

Flow cytometry demonstrated that the apoptosis or death of EV71-infected RD cells was increased by 37.1% with a multiplicity of infection (MOI) of 5 at 20 h postinfection. The production of IL-4, IL-10 and TNF-α was enhanced by the subsequent EV71 infection. PCR array revealed significant changes in the expressions of apoptotic genes. Among 84 genes, 42 genes were down-regulated after EV71 infection at 8 h, whereas 32 genes were up-regulated at 20 h postinfection. Moreover, the ligands of TNF superfamily such as FasL, CD40L and TNF-α were significantly up-regulated and enhanced the expressions of apoptosis-related cysteine peptidases, including caspase-10, -8, -7 and -3. In addition, EV71 infection induces the phosphorylation of AKT2, JNK1/2, c-Jun and NF-κB at 20 h postinfection.

Conclusion

PCR array for the determination of apoptosis gene expressions is an informative assay in elucidating biological pathways. During the early stage of EV71 infection, the apoptotic process of RD cells is significantly delayed. EV71 infection can also induce the expressions of FasL, TNF-α and CD40L, which contribute to the apoptosis of RD cells.

Keywords:
Enterovirus 71; PCR array; FasL; CD40L; Apoptosis