Differential protein expression between EBV-positive and negative epithelial cells

Haibo Yu,Guiyuan Li,Jianhong Lu,Lian Zhao,Zhengyuan Yu,Shourong Sheng,Zhaojian Gong,Xiaoling Li,Lielian Zuo,Qijia Yan,Wei Xiong

doi:10.4236/jbpc.2013.42011

Abstract

Epstein Barr virus infection is believed to play a role in the development of nasopharyngeal carcinoma. In order to investigate the function of EBV in epithelial cell, proteomic methods were used to find and identify the differential proteins and expected to elucidate the mechanism of EBV. Altered protein expressions were found between 293 cell (HEK293) and EBV infected cell (293-EBV). In this study, we separated differential expressed proteins using 2D-DIGE method while matrix-assisted laser desorption/ionization tandem time of flight mass spectrometry (MALDI-TOF-MS) method was used to identify proteins. The results showed that 14 proteins were up regulated and 3 proteins were down regulated in 293-EBV cells. Bioinformatic analysis showed that these proteins are involved in cell proliferation, metastasis, apoptosis, metabolism, and signal transduction. Western blotting analysis was further carried out to verify the MS results. Thus, EBV may exert its functions by mediating differential expression of these proteins.

Highlights

Epstein Barr virus infection has been believed to play a key role in the development of many tumors such as
Nasopharyngeal carcinoma (NPC) which prevalently accrus in southern China and Southeast Asia
We obtained the images of proteins from 293 cells, 293-EBV cells and from the internal standard using different emission filters on the Typhoon 9400 fluorescence scanner

Summary

Introduction

Epstein Barr virus infection has been believed to play a key role in the development of many tumors such as nasopharyngeal carcinoma (NPC) which prevalently accrus in southern China and Southeast Asia. The proliferation rate of epithelial cell was faster after transfection with EBV genome. The mechanism of EBV is still unclear, in this work powerful proteomic technologies were used to elucidate the potential roles of EBV. As it is known, protein is the ultimate life performer. How does EBV regulate the protein profile of epithelial cell? Our study uses the differential in-gel electrophoresis DIGE and MALDI-TOF-MS(Matrix-assisted laser desorption/ionization time of flight mass spectrometry) to select and identify differential expressed protein compared 293 cells with 293-EBV cells. The results are analyzed to illuminate the mechanism of EBV in promoting cell proliferation and differentiation in protein level, which supply a new target and clue for cure and prognostic of EBV associated cancer

Methods

Results

Conclusion