Basic Research
Copyright ©The Author(s) 2003. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Dec 15, 2003; 9(12): 2726-2731
Published online Dec 15, 2003. doi: 10.3748/wjg.v9.i12.2726
Differentially expressed proteins of gamma-ray irradiated mouse intestinal epithelial cells by two-dimensional electrophoresis and MALDI-TOF mass spectrometry
Bo Zhang, Yong-Ping Su, Guo-Ping Ai, Xiao-Hong Liu, Feng-Chao Wang, Tian-Min Cheng
Bo Zhang, Yong-Ping Su, Guo-Ping Ai, Xiao-Hong Liu, Feng-Chao Wang, Tian-Min Cheng, Institute of Combined Injury of PLA, Third Military Medical University, Chongqing 400038, China
Author contributions: All authors contributed equally to the work.
Supported by the National Natural Science Foundation of China, No.30230360
Correspondence to: Professor Yong-Ping Su, Institute of Combined Injury of PLA, Third Military Medical University, Gaotanyan Street 30, Chongqing 400038, China.
Telephone: +86-23-68752355 Fax: +86-23-68752279
Received: May 13, 2003
Revised: May 23, 2003
Accepted: June 12, 2003
Published online: December 15, 2003

AIM: To identify the differentially expressed proteins involved in ionizing radiation in mice and to explore new ways for studying radiation-related proteins.

METHODS: Bal B/c mice grouped as sham-irradiation, 3 h and 72 h irradiation were exposed to 9.0Gy single dose of γ-irradiation. Intestinal epithelia were isolated from mice, and total proteins were extracted with urea containing solution. A series of methods were used, including two-dimensional electrophoresis, PDQuest 2-DE software analysis, peptide mass fingerprinting based on matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) and SWISS-PROT database searching, to separate and identify the differential proteins. Western blotting and RT-PCR were used to validate the differentially expressed proteins.

RESULTS: Mouse intestine was severely damaged by 9.0 Gy γ-irradiation. Image analysis of two-dimensional gels revealed that averages of 638 ± 39, 566 ± 32 and 591 ± 29 protein spots were detected in 3 groups, respectively, and the majority of these protein spots were matched. About 360 protein spots were matched between normal group and 3 h irradiation group, and the correlation coefficient was 0.78 by correlation analysis of gels. Also 312 protein spots matched between normal group and 72 h irradiation group, and 282 protein spots between 3 h and 72 h irradiation groups. Twenty-eight differential protein spots were isolated from gels, digested with trypsin, and measured with MALDI-TOF-MS. A total of 25 spots yielded good spectra, and 19 spots matched known proteins after database searching. These proteins were mainly involved in anti-oxidation, metabolism, signal transduction, and protein post-translational processes. Western-blotting confirmed that enolase was up-regulated by γ-irradiation. Up-regulation of peroxiredoxin I was verified by applying RT-PCR technique, but no change occurred in Q8VC72.

CONCLUSION: These differentially expressed proteins might play important roles when mouse intestine was severely injured by γ-irradiation. It is suggested that differential proteomic analysis may be a useful tool to study the proteins involved in radiation damage of mouse intestinal epithelia.

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