標題: An Informatics-assisted Label-free Approach for Personalized Tissue Membrane Proteomics: Case Study on Colorectal Cancer
作者: Han, Chia-Li
Chen, Jinn-Shiun
Chan, Err-Cheng
Wu, Chien-Peng
Yu, Kun-Hsing
Chen, Kuei-Tien
Tsou, Chih-Chiang
Tsai, Chia-Feng
Chien, Chih-Wei
Kuo, Yung-Bin
Lin, Pei-Yi
Yu, Jau-Song
Hsueh, Chuen
Chen, Min-Chi
Chan, Chung-Chuan
Chang, Yu-Sun
Chen, Yu-Ju
生物科技學院
College of Biological Science and Technology
公開日期: 1-Apr-2011
摘要: We developed a multiplexed label-free quantification strategy, which integrates an efficient gel-assisted digestion protocol, high-performance liquid chromatography tandem MS analysis, and a bioinformatics alignment method to determine personalized proteomic profiles for membrane proteins in human tissues. This strategy provided accurate (6% error) and reproducible (34% relative S. D.) quantification of three independently purified membrane fractions from the same human colorectal cancer (CRC) tissue. Using CRC as a model, we constructed the personalized membrane protein atlas of paired tumor and adjacent normal tissues from 28 patients with different stages of CRC. Without fractionation, this strategy confidently quantified 856 proteins (>= 2 unique peptides) across different patients, including the first and robust detection (Mascot score: 22,074) of the well-documented CRC marker, carcinoembryonic antigen 5 by a discovery-type proteomics approach. Further validation of a panel of proteins, annexin A4, neutrophils defensin A1, and claudin 3, confirmed differential expression levels and high occurrences (48-70%) in 60 CRC patients. The most significant discovery is the overexpression of stomatin-like 2 (STOML2) for early diagnostic and prognostic potential. Increased expression of STOML2 was associated with decreased CRC-related survival; the mean survival period was 34.77 +/- 2.03 months in patients with high STOML2 expression, whereas 53.67 +/- 3.46 months was obtained for patients with low STOML2 expression. Further analysis by ELISA verified that plasma concentrations of STOML2 in early-stage CRC patients were elevated as compared with those of healthy individuals (p < 0.001), suggesting that STOML2 may be a noninvasive serological biomarker for early CRC diagnosis. The overall sensitivity of STOML2 for CRC detection was 71%, which increased to 87% when combined with CEA measurements. This study demonstrated a sensitive, label-free strategy for differential analysis of tissue membrane proteome, which may provide a roadmap for the subsequent identification of molecular target candidates of multiple cancer types. Molecular & Cellular Proteomics 10: 10.1074/mcp.M110.003087, 1-15, 2011.
URI: http://dx.doi.org/10.1074/mcp.M110.003087
http://hdl.handle.net/11536/9080
ISSN: 1535-9476
DOI: 10.1074/mcp.M110.003087
期刊: MOLECULAR & CELLULAR PROTEOMICS
Volume: 10
Issue: 4
結束頁: 
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