Basic Research
Copyright ©2008 The WJG Press and Baishideng. All rights reserved.
World J Gastroenterol. Jan 28, 2008; 14(4): 554-562
Published online Jan 28, 2008. doi: 10.3748/wjg.14.554
Discovering differential protein expression caused by CagA-induced ERK pathway activation in AGS cells using the SELDI-ProteinChip platform
Zhen Ge, Yong-Liang Zhu, Xian Zhong, Jie-Kai Yu, Shu Zheng
Zhen Ge, Xian Zhong, Jie-Kai Yu, Shu Zheng, Cancer Institute, Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou 310009, Zhejiang Province, China
Yong-Liang Zhu, Gastroenterological Department, Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou 310009, Zhejiang Province, China
Zhen Ge, James D Watson, Institute of Genome Sciences, College of Life Sciences, Zhejiang University, Hangzhou 310008, Zhejiang Province, China
Correspondence to: Professor Shu Zheng, 88 Jiefang Road, Cancer Institute of Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou 310009, Zhejiang, China. zhengshu@zju.edu.cn
Telephone: +86-571-87784501
Fax: +86-571-87214404
Received: August 25, 2007
Revised: October 8, 2007
Published online: January 28, 2008
Abstract

AIM: To identify the protein expression differences related to the CagA-induced ERK pathway activation in AGS cells.

METHODS: Human AGS cells transfected with cagA and blank vector were treated with specific mitogen-activated protein kinase kinase (MEK) inhibitor. Total cell proteins were combined by strong anion exchange (SAX2) and weak cation exchange (CM10) ProteinChip arrays and analyzed using surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF-MS) proteomics technology. Protein expression profiles were compared with those of inhibitor-untreated cagA transfectants. SwissProt/TrEMBL database searching for differentially expressed proteins was carried out using the TagIdent tool with the pI and mass information.

RESULTS: When a total of 16 proteins that showed expression differences in inhibitor-untreated cagA transfectants were compared with vector transfectants, three proteins with m/z 4229, 8162 and 9084 were found to have no expression differences after treatment with MEK inhibitor, while the other 13 maintained the same expression differences after inhibitor treatment. Seven pieces of meaningful matching information for the three proteins were obtained from database searching.

CONCLUSION: Biomarkers with m/z 4229, 8162 and 9084 are ERK1/2 phosphorylation dependent, and therefore are the downstream molecules of ERK1/2 in the ERK/MAPK signaling pathway. The three biomarkers may be important cancer-associated proteins according to SwissProt/TrEMBL database information.

Keywords: CagA; ERK pathway; SELDI-TOF-MS; ProteinChip