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Urine expression of TIMP-1, serpinB1 and seminogelin 2 may differentiate those men with low risk or no evidence of prostate cancer from men with high risk or metastatic disease
Adam S. Feldman, MD, MPH, Sarah Prophet, BA, Mary Fergus, BA, Bruce Zetter, PhD.
Massachusetts General Hospital, Boston, MA, USA.

BACKGROUND - While serum PSA and other available diagnostic biomarkers can provide valuable guidance for assessing the risk of prostate cancer (PCa), limitations in accuracy persist. Novel biomarkers with improved performance characteristics are needed. Using mass spectrometry-based proteomics, we have identified three urinary proteins with significantly different expression patterns across PCa stages. This study evaluates the expression of these potential biomarkers in urine.
METHODS - Urinary protein concentrations of three proteins, TIMP1, serpinB1, and semenogelin 2, were assessed via Western blot and ELISA for 160 total urine samples. Each patient group (control, Gleason 6 PCa, Gleason ≥8 PCa, and metastatic PCa) had 40 samples. Urine protein was isolated using Amicon Ultra-15 Centrifugal Filter Units for Western blotting. ELISAs were performed using untreated raw urine samples. Immunohistochemistry (IHC) was performed on prostate tissue sections for all three proteins of interest.
RESULTS - TIMP1 levels were statistically higher in control and Gleason 6 PCa urine samples than for Gleason ≥8 and metastatic disease (2.19 ± 1.7 vs. 1.23 ± 1.13 ng/mL; p = 0.002). Expression of serpinB1 was significantly higher in men with Gleason 6 PCa than those with high-grade Gleason ≥8 PCa (0.71 ± 0.49 vs. 0.23 ± 0.33 ng/mL; p = 0.003). Metastatic PCa had significantly higher semenogelin2 concentrations in urine than healthy men (155.68 ± 74.3 vs. 81.55 ± 55.6 pg/mL; p = 0.02), and expression levels seem to rise with disease progression. IHC staining of tissue sections corroborated these findings.
CONCLUSIONS - Our results indicate differences in urinary concentrations of TIMP1, serpinB1, and semenogelin 2 across PCa stages. These novel biomarkers allowed distiction between men without prostate cancer or low-risk disease and those with high-risk or metastatic disease. These proteins represent potentially valuable non-invasive prostate cancer biomarkers and warrant further investigation.


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