Identification and validation of potential new biomarkers for prostate cancer diagnosis and prognosis using 2D-DIGE and MS.

Cordelia Geisler,Till Braunschweig,Glen Kristiansen,David Pfister,Helmut E Meyer,Axel Heidenreich,Corinna Henkel,Christoph H Borchers,Susanne Fuessel,Hanna C Diehl,Birte Beine,Angela M Jackson,Sonja Gostek,Nadine T Gaisa,Ruth Knüchel

doi:10.1155/2015/454256

Abstract

This study was designed to identify and validate potential new biomarkers for prostate cancer and to distinguish patients with and without biochemical relapse. Prostate tissue samples analyzed by 2D-DIGE (two-dimensional difference in gel electrophoresis) and mass spectrometry (MS) revealed downregulation of secernin-1 (P < 0.044) in prostate cancer, while vinculin showed significant upregulation (P < 0.001). Secernin-1 overexpression in prostate tissue was validated using Western blot and immunohistochemistry while vinculin expression was validated using immunohistochemistry. These findings indicate that secernin-1 and vinculin are potential new tissue biomarkers for prostate cancer diagnosis and prognosis, respectively. For validation, protein levels in urine were also examined by Western blot analysis. Urinary vinculin levels in prostate cancer patients were significantly higher than in urine from nontumor patients (P = 0.006). Using multiple reaction monitoring-MS (MRM-MS) analysis, prostatic acid phosphatase (PAP) showed significant higher levels in the urine of prostate cancer patients compared to controls (P = 0.012), while galectin-3 showed significant lower levels in the urine of prostate cancer patients with biochemical relapse, compared to those without relapse (P = 0.017). Three proteins were successfully differentiated between patients with and without prostate cancer and patients with and without relapse by using MRM. Thus, this technique shows promise for implementation as a noninvasive clinical diagnostic technique.

Highlights

Prostate cancer is the most commonly occurring cancer among men in economically developed countries
For the identification of differentially regulated proteins in prostate cancer, 12 prostatectomy samples from prostate cancer patients without biochemical relapse, 11 prostatectomy samples from patients with biochemical relapse, and 14 corresponding tumor-free prostate cancer tissue samples were comparatively analyzed by 2D-DIGE saturation labeling
2D-DIGE combined with mass spectrometry (MS) is a labor-intensive and expensive method with a low throughput of the 2D-DIGE

Summary

Introduction

Prostate cancer is the most commonly occurring cancer among men in economically developed countries. 248,500 men died of prostate cancer in 2008 [1], most men diagnosed with prostate cancer die from causes other than prostate cancer [2]. Some prostate cancers are clinically relevant from the start, while others will acquire clinical significance over the years [3, 4]. High-grade prostatic intraepithelial neoplasia often develops into prostate cancer [5,6,7], many prostate cancers may remain indolent for 10–15 years or longer [8]. Almost 100% of patients who show a biochemical relapse will later develop a clinical relapse [11], with metastasis causing death [12, 13]

Results

Discussion

Conclusion