Characterization of the Human Gastric Fluid Proteome Reveals Distinct pH-Dependent Protein Profiles: Implications for Biomarker Studies

被引：18

作者：

Kam, Siok Yuen ^{[1
]}

Hennessy, Thomas ^{[2
]}

Chua, Seow Ching ^{[1
]}

Gan, Chee Sian ^{[2
]}

Philp, Robin ^{[2
]}

Hon, Ka Ka ^{[1
]}

Lai, Liyun ^{[1
]}

Chan, Weng Hoong ^{[3
]}

Ong, Hock Soo ^{[3
]}

Wong, Wai Keong ^{[3
]}

Lim, Kiat Hon ^{[4
]}

Ling, Khoon Lin ^{[5
]}

Tan, Hwee Sian ^{[2
]}

Tan, Mei Mei ^{[2
]}

Ho, Mengfatt ^{[1
]}

Kon, Oi Lian ^{[1
]}

机构：

[1] Natl Canc Ctr, Div Med Sci, Humphrey Oei Inst Canc Res, Singapore 169610, Singapore

[2] Agilent Technol, Life Sci & Chem Anal Grp, Singapore, Singapore

[3] Singapore Gen Hosp, Dept Gen Surg, Singapore 0316, Singapore

[4] Singapore Gen Hosp, Dept Pathol, Singapore 0316, Singapore

[5] Singapore Gen Hosp, Dept Gastroenterol, Singapore 0316, Singapore

来源：

JOURNAL OF PROTEOME RESEARCH | 2011年 / 10卷 / 10期

关键词：

gastric juice; gastric cancer; biomarkers; two-dimensional gel electrophoresis (2-DE); HPLC-Chip/MS; multidimensional LC-MS/MS; mass spectrometry; ELECTROPHORETIC PATTERN; CANCER; JUICE; DIAGNOSIS; EXPRESSION; ALPHA-1-ANTITRYPSIN; IDENTIFICATION; SPECTROMETRY; COMPONENTS; SECRETION;

D O I：

10.1021/pr200349z

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Gastric fluid is a source of gastric cancer biomarkers. However, very little is known about the normal gastric fluid proteome and its biological variations. In this study, we performed a comprehensive analysis of the human gastric fluid proteome using samples obtained from individuals with benign gastric conditions. Gastric fluid proteins were prefractionated using ultracentrifuge filters (3 kDa cutoff) and analyzed by two-dimensional gel electrophoresis (2-DE) and multidimensional LC-MS/MS. Our 2-DE analysis of 170 gastric fluid samples revealed distinct protein profiles for acidic and neutral samples, highlighting pH effects on protein composition. By 2D LC-MS/MS analysis of pooled samples, we identified 284 and 347 proteins in acidic and neutral samples respectively (FDR <= 1%), of which 265 proteins (72.4%) overlapped. However, unlike neutral samples, most proteins in acidic samples were identified from peptides in the filtrate (i.e., <3 kDa). Consistent with this finding, immunoblot analysis of six potential gastric cancer biomarkers rarely detected full-length proteins in acidic samples. These findings have important implications for biomarker studies because a majority of gastric cancer patients have neutral gastric fluid compared to noncancer controls. Consequently, sample stratification, choice of proteomic approaches, and validation strategy can profoundly affect the interpretation of biomarker findings. These observations should help to refine gastric fluid biomarker studies.

引用

页码：4535 / 4546

页数：12

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