Identification of Differentially Expressed Proteins in Direct Expressed Prostatic Secretions of Men with Organ-confined Versus Extracapsular Prostate Cancer

被引：75

作者：

Kim, Yunee ^{[1
]}

Ignatchenko, Vladimir ^{[2
]}

Yao, Cindy Q. ^{[1
,4
]}

Kalatskaya, Irina ^{[4
]}

Nyalwidhe, Julius O. ^{[3
,5
,6
]}

Lance, Raymond S. ^{[3
,6
,7
]}

Gramolini, Anthony O. ^{[8
]}

Troyer, Dean A. ^{[3
,5
,6
]}

Stein, Lincoln D. ^{[4
]}

Boutros, Paul C. ^{[4
]}

Medin, Jeffrey A. ^{[1
,2
,9
]}

Semmes, O. John ^{[3
,5
,6
]}

Drake, Richard R. ^{[3
,5
,6
,10
]}

Kislinger, Thomas ^{[1
,2
]}

机构：

[1] Univ Toronto, Dept Med Biophys, Toronto, ON, Canada

[2] Univ Hlth Network, Ontario Canc Inst, Toronto, ON M5R 2W7, Canada

[3] Univ Hlth Network, Ontario Canc Inst, Toronto, ON M5R 2W7, Canada

[4] Ontario Inst Canc Res, Toronto, ON, Canada

[5] Eastern Virginia Med Sch, Dept Microbiol & Mol Cell Biol, Norfolk, VA 23501 USA

[6] Eastern Virginia Med Sch, Leroy T Canoles Jr Canc Res Ctr, Norfolk, VA 23501 USA

[7] Eastern Virginia Med Sch, Dept Urol, Norfolk, VA 23501 USA

[8] Univ Toronto, Dept Physiol, Toronto, ON, Canada

[9] Univ Toronto, Inst Med Sci, Toronto, ON M5S 1A1, Canada

[10] Med Univ S Carolina, Dept Cell & Mol Pharmacol & Expt Therapeut, Charleston, SC 29425 USA

来源：

MOLECULAR & CELLULAR PROTEOMICS | 2012年 / 11卷 / 12期

基金：

美国国家卫生研究院;

关键词：

RADICAL RETROPUBIC PROSTATECTOMY; BIOMARKER DISCOVERY; PROTEOMIC ANALYSIS; MASS-SPECTROMETRY; BIOCHEMICAL RECURRENCE; NEUTRAL ENDOPEPTIDASE; SHOTGUN PROTEOMICS; HUMAN URINE; LARGE-SCALE; OVARIAN;

D O I：

10.1074/mcp.M112.017889

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Current protocols for the screening of prostate cancer cannot accurately discriminate clinically indolent tumors from more aggressive ones. One reliable indicator of outcome has been the determination of organ-confined versus nonorgan-confined disease but even this determination is often only made following prostatectomy. This underscores the need to explore alternate avenues to enhance outcome prediction of prostate cancer patients. Fluids that are proximal to the prostate, such as expressed prostatic secretions (EPS), are attractive sources of potential prostate cancer biomarkers as these fluids likely bathe the tumor. Direct-EPS samples from 16 individuals with extracapsular (n = 8) or organ-confined (n = 8) prostate cancer were used as a discovery cohort, and were analyzed in duplicate by a nine-step MudPIT on a LTQ-Orbitrap XL mass spectrometer. A total of 624 unique proteins were identified by at least two unique peptides with a 0.2% false discovery rate. A semiquantitative spectral counting algorithm identified 133 significantly differentially expressed proteins in the discovery cohort. Integrative data mining prioritized 14 candidates, including two known prostate cancer biomarkers: prostate-specific antigen and prostatic acid phosphatase, which were significantly elevated in the direct-EPS from the organ-con-fined cancer group. These and five other candidates (SFN, MME, PARK7, TIMP1, and TGM4) were verified by Western blotting in an independent set of direct-EPS from patients with biochemically recurrent disease (n = 5) versus patients with no evidence of recurrence upon follow-up (n = 10). Lastly, we performed proof-of-concept SRM-MS-based relative quantification of the five candidates using unpurified heavy isotope-labeled synthetic peptides spiked into pools of EPS-urines from men with extracapsular and organ-confined prostate tumors. This study represents the first efforts to define the direct-EPS proteome from two major subclasses of prostate cancer using shotgun proteomics and verification in EPS-urine by SRM-MS. Molecular & Cellular Proteomics 11: 10.1074/mcp.M112.017889, 1870-1884, 2012.

引用

页码：1870 / 1884

页数：15

共 83 条

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