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Sci Rep. 2016 Sep 30;6:34446. doi: 10.1038/srep34446.

Proteome Profiling of Urinary Exosomes Identifies Alpha 1-Antitrypsin and H2B1K as Diagnostic and Prognostic Biomarkers for Urothelial Carcinoma.

Lin SY1,2,3, Chang CH4, Wu HC4, Lin CC1,5, Chang KP6, Yang CR4, Huang CP1,4, Hsu WH1,2,7, Chang CT1,2,3, Chen CJ8,9.

Author information

1
Institute of Clinical Medical Science, China Medical University College of Medicine, Taichung, Taiwan.
2
Department of Internal Medicine, China Medical University Hospital, Taichung, Taiwan.
3
Division of Nephrology and Kidney Institute, China Medical University Hospital, Taichung, Taiwan.
4
Department of Urology, China Medical University Hospital, Taichung, Taiwan.
5
Department of Hematology and Oncology, China Medical University Hospital, Taichung, Taiwan.
6
Department of Pathology, China Medical University Hospital, Taichung, Taiwan.
7
Department of Chest Medicine, China Medical University Hospital, Taichung, Taiwan.
8
Graduate Institute of Integrated Medicine, China Medical University, Taichung, Taiwan.
9
Proteomics Core Laboratory, Department of Medical Research, China Medical University Hospital, Taichung, Taiwan.

Abstract

MALDI-TOF spectrometry has not been used for urinary exosome analysis. We used it for determining UC biomarkers. From 2012 to 2015, we enrolled 129 consecutive patients with UC and 62 participants without UC. Exosomes from their urine were isolated, and analyzed through MALDI-TOF spectrometry. Immunohistochemical (IHC) analysis of another 122 UC and 26 non-UC tissues was conducted to verify the discovered biomarkers. Two peaks at m/z 5593 (fragmented peptide of alpha-1-antitrypsin; sensitivity, 50.4%; specificity, 96.9%) and m/z 5947 (fragmented peptide of histone H2B1K sensitivity, 62.0%; specificity, 92.3%) were identified as UC diagnosis exosome biomarkers. UC patients with detectable histone H2B1K showed 2.29- and 3.11-fold increased risks of recurrence and progression, respectively, compared with those with nondetectable histone H2B1K. Verification results of IHC staining revealed significantly higher expression of alpha 1-antitrypsin (p = 0.038) and H2B1K (p = 0.005) in UC tissues than in normal tissues. The expression of alpha 1-antitrypsin and H2B1K in UC tissues was significantly correlated with UC grades (p < 0.05). Urinary exosome proteins alpha 1-antitrypsin and histone H2B1K, which are identified through MALDI-TOF analysis, could facilitate rapid diagnosis and prognosis of UC.

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