Comparative and targeted proteomic analyses of urinary microparticles from bladder cancer and hernia patients.
Bladder cancer is a common urologic cancer whose incidence continues to rise annually. Urinary microparticles are an attractive material for noninvasive bladder cancer biomarker discovery. In this study, we applied isotopic dimethylation labeling coupled with liquid chromatography-tandem mass spectrometry (LC-MS/MS) to discover bladder cancer biomarkers in urinary microparticles isolated from hernia (control) and bladder cancer patients. This approach identified 2964 proteins based on more than two distinct peptides, of which 2058 had not previously been reported as constituents of human urine exosomes/microparticles. A total of 107 differentially expressed proteins were identified as candidate biomarkers. Differences in the concentrations of 29 proteins (41 signature peptides) were precisely quantified by LC-MRM/MS in 48 urine samples of bladder cancer, hernia, and urinary tract infection/hematuria. Concentrations of 24 proteins changed significantly (p<0.05) between bladder cancer (n=28) and hernia (n=12), with area-under-the-curve values ranging from 0.702 to 0.896. Finally, we quantified tumor-associated calcium-signal transducer 2 (TACSTD2) in raw urine specimens (n=221) using a commercial ELISA and confirmed its potential value for diagnosis of bladder cancer. Our study reveals a strong association of TACSTD2 with bladder cancer and highlights the potential of human urinary microparticles in the noninvasive diagnosis of bladder cancer.
Chang Gung Bioinformatics Center, Department of Urology, Chang Gung Memorial Hospital, and Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan 333, Taiwan.
Chien-Lun
CL
Chen
Yue-Fan
YF
Lai
Petrus
P
Tang
Kun-Yi
KY
Chien
Jau-Song
JS
Yu
Cheng-Han
CH
Tsai
Hsiao-Wei
HW
Chen
Chih-Ching
CC
Wu
Ting
T
Chung
Chia-Wei
CW
Hsu
Chi-De
CD
Chen
Yu-Sun
YS
Chang
Phei-Lang
PL
Chang
Yi-Ting
YT
Chen
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000595:Amino Acid Sequence; D000951:Antigens, Neoplasm / Q000652:urine; D019540:Area Under Curve; D014408:Biomarkers, Tumor / Q000652:urine*; D016022:Case-Control Studies; D015815:Cell Adhesion Molecules / Q000652:urine; D002853:Chromatography, Liquid / Q000379:methods; D004797:Enzyme-Linked Immunosorbent Assay; D055354:Exosomes / Q000737:chemistry*; D005260:Female; D006417:Hematuria / Q000175:diagnosis; D006547:Hernia / Q000175:diagnosis; D006801:Humans; D007553:Isotope Labeling; D008297:Male; D013058:Mass Spectrometry / Q000379:methods; D008875:Middle Aged; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins / Q000652:urine; D020543:Proteome / Q000032:analysis*; D040901:Proteomics / Q000379:methods; D015203:Reproducibility of Results; D001749:Urinary Bladder Neoplasms / Q000737:chemistry / Q000175:diagnosis*
D000951:Antigens, Neoplasm; D014408:Biomarkers, Tumor; D015815:Cell Adhesion Molecules; D009363:Neoplasm Proteins; D020543:Proteome; C499380:TACSTD2 protein, human