title: Comparative Tissue Proteomics of Microdissected Specimens Reveals Novel Candidate Biomarkers of Bladder Cancer.
abstract: More than 380,000 new cases of bladder cancer are diagnosed worldwide, accounting for ∼150,200 deaths each year. To discover potential biomarkers of bladder cancer, we employed a strategy combining laser microdissection, isobaric tags for relative and absolute quantitation labeling, and liquid chromatography-tandem MS (LC-MS/MS) analysis to profile proteomic changes in fresh-frozen bladder tumor specimens. Cellular proteins from four pairs of surgically resected primary bladder cancer tumor and adjacent nontumorous tissue were extracted for use in two batches of isobaric tags for relative and absolute quantitation experiments, which identified a total of 3220 proteins. A DAVID (database for annotation, visualization and integrated discovery) analysis of dysregulated proteins revealed that the three top-ranking biological processes were extracellular matrix organization, extracellular structure organization, and oxidation-reduction. Biological processes including response to organic substances, response to metal ions, and response to inorganic substances were highlighted by up-expressed proteins in bladder cancer. Seven differentially expressed proteins were selected as potential bladder cancer biomarkers for further verification. Immunohistochemical analyses showed significantly elevated levels of three proteins-SLC3A2, STMN1, and TAGLN2-in tumor cells compared with noncancerous bladder epithelial cells, and suggested that TAGLN2 could be a useful tumor tissue marker for diagnosis (AUC = 0.999) and evaluating lymph node metastasis in bladder cancer patients. ELISA results revealed significantly increased urinary levels of both STMN1 and TAGLN2 in bladder cancer subgroups compared with control groups. In comparisons with age-matched hernia urine specimens, urinary TAGLN2 in bladder cancer samples showed the largest fold change (7.13-fold), with an area-under-the-curve value of 0.70 (p < 0.001, n = 205). Overall, TAGLN2 showed the most significant overexpression in individual bladder cancer tissues and urine specimens, and thus represents a potential biomarker for noninvasive screening for bladder cancer. Our findings highlight the value of bladder tissue proteome in providing valuable information for future validation studies of potential biomarkers in urothelial carcinoma.
authors.affiliation: From the ‡Department of Urology, Chang Gung Memorial Hospital, Taoyuan, Taiwan; §School of Medicine, College of Medicine, Chang Gung University, Taoyuan, Taiwan;
authors.forename: Chien-Lun
authors.initials: CL
authors.lastname: Chen
authors.affiliation: ¶Molecular Medicine Research Center, Chang Gung University, Taoyuan, Taiwan;
authors.forename: Ting
authors.initials: T
authors.lastname: Chung
authors.affiliation: ¶Molecular Medicine Research Center, Chang Gung University, Taoyuan, Taiwan; ‖Department of Medical Biotechnology and Laboratory Science, College of Medicine, Chang Gung University, Taoyuan, Taiwan;
authors.forename: Chih-Ching
authors.initials: CC
authors.lastname: Wu
authors.affiliation: **Department of Pathology, Chang Gung Memorial Hospital, Taoyuan, Taiwan;
authors.forename: Kwai-Fong
authors.initials: KF
authors.lastname: Ng
authors.affiliation: ¶Molecular Medicine Research Center, Chang Gung University, Taoyuan, Taiwan; ‡‡Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan, Taiwan;
authors.forename: Jau-Song
authors.initials: JS
authors.lastname: Yu
authors.affiliation: ‡‡Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan, Taiwan;
authors.forename: Cheng-Han
authors.initials: CH
authors.lastname: Tsai
authors.affiliation: ¶Molecular Medicine Research Center, Chang Gung University, Taoyuan, Taiwan; ‡‡Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan, Taiwan;
authors.forename: Yu-Sun
authors.initials: YS
authors.lastname: Chang
authors.affiliation: ¶Molecular Medicine Research Center, Chang Gung University, Taoyuan, Taiwan;
authors.forename: Ying
authors.initials: Y
authors.lastname: Liang
authors.affiliation: From the ‡Department of Urology, Chang Gung Memorial Hospital, Taoyuan, Taiwan; §School of Medicine, College of Medicine, Chang Gung University, Taoyuan, Taiwan;
authors.forename: Ke-Hung
authors.initials: KH
authors.lastname: Tsui
authors.affiliation: ¶Molecular Medicine Research Center, Chang Gung University, Taoyuan, Taiwan; ‡‡Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan, Taiwan; §§Department of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan, Taiwan ytchen@mail.cgu.edu.tw.
authors.forename: Yi-Ting
authors.initials: YT
authors.lastname: Chen
mesh_terms: D000368:Aged; D014408:Biomarkers, Tumor / Q000378:metabolism* / Q000652:urine; D002853:Chromatography, Liquid; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D008297:Male; D042282:Microdissection; D008840:Microfilament Proteins / Q000378:metabolism* / Q000652:urine; D008875:Middle Aged; D009124:Muscle Proteins / Q000378:metabolism* / Q000652:urine; D040901:Proteomics / Q000379:methods*; D050777:Stathmin / Q000378:metabolism* / Q000652:urine; D053719:Tandem Mass Spectrometry; D001749:Urinary Bladder Neoplasms / Q000378:metabolism* / Q000652:urine
chemical_list: D014408:Biomarkers, Tumor; D008840:Microfilament Proteins; D009124:Muscle Proteins; C493995:STMN1 protein, human; D050777:Stathmin; C585387:Tagln2 protein, human