Searching Urinary Tumor Markers for Bladder Cancer Using a Two-Dimensional Differential Gel Electrophoresis (2D-DIGE) Approach

Aiming at identifying biomarkers for bladder cancer, the urinary proteome was explored through a two-dimensional gel-based proteomic approach (2D-DIGE) coupled with mass spectrometry and database interrogation. The increased expression of proteins differentially expressed between patients with bladder tumors and controls such as Reg-1 and keratin 10 was confirmed to be associated with bladder cancer progression on bladder cancer cell lines by immunoblotting, and bladder tumors by immunohistochemistry. Moreover, the association of these proteins, especially Reg-1, with tumor staging and clinical outcome was confirmed by immunohistochemistry using an independent series of bladder tumors contained in tissue microarrays (n = 292). Furthermore, Reg-1 was quantified using an independent series of urinary specimens (n = 80) and provided diagnostic utility to discriminate patients with bladder cancer and controls (area under the curve (AUC = 0.88)). Thus, the 2D-DIGE approach has identified Reg-1 as a biomarker for bladder cancer diagnostics, staging, and outcome prognosis. Keywords: 2D-DIGE • bladder cancer • MALDI-TOF−TOF • Reg-1