Identification of Clinically Relevant Protein Targets in Prostate Cancer with 2D-DIGE Coupled Mass Spectrometry and Systems Biology Network Platform

Abstract

Prostate cancer (PCa) is the most common type of cancer found in men and among the leading causes of cancer death in the western world. In the present study, we compared the individual protein expression patterns from histologically characterized PCa and the surrounding benign tissue obtained by manual micro dissection using highly sensitive two-dimensional differential gel electrophoresis (2D-DIGE) coupled with mass spectrometry. Proteomic data revealed 118 protein spots to be differentially expressed in cancer (n = 24) compared to benign (n = 21) prostate tissue. These spots were analysed by MALDI-TOF-MS/MS and 79 different proteins were identified. Using principal component analysis we could clearly separate tumor and normal tissue and two distinct tumor groups based on the protein expression pattern. By using a systems biology approach, we could map many of these proteins both into major pathways involved in PCa progression as well as into a group of potential diagnostic and/or prognostic markers. Due to complexity of the highly interconnected shortest pathway network, the functional sub networks revealed some of the potential candidate biomarker proteins for further validation. By using a systems biology approach, our study revealed novel proteins and molecular networks with altered expression in PCa. Further functional validation of individual proteins is ongoing and might provide new insights in PCa progression potentially leading to the design of novel diagnostic and therapeutic strategies.

Ubiquitin carboxyl-terminal hydrolase 1 (UCHL1) is a potential tumour suppressor in prostate cancer and is frequently silenced by promoter methylation

“…Our proteomic analysis of prostate tissue samples highlighted significant downregulation of UCHL1 in cancer compared to the surrounding benign tissue [3]. In the current study, we analyzed whether downregulation of UCHL1 in PCa is due to promoter hypermethylation.…”

From a 2DE-Gel Spot to Protein Function: Lesson Learned From HS1 in Chronic Lymphocytic Leukemia

“…The aim of this manuscript is to describe an optimized protocol for the identification of molecules involved in the pathogenesis of CLL, even if this approach can be extended to other diseases and/or other sample types [16][17][18] . By comparing the proteomic profiles of 2 subsets of CLL patients, good vs bad prognosis 19 , we demonstrated that they differ in the phosphorylation status of HS1 and that its activation affects the migration and adhesion capacity of leukemic cells.…”

The proteomics in prostate cancer biomarker discovery

“…2D-DIGE c протеомным MS-анализом образ-цов ткани РПЖ в сравнении с нормальной тканью тех же самых пациентов показал, что дифференциаль-но экспрессируемые белки (ДЭБ) в этих 2 группах тканей вовлечены в развитие опухолевого процесса и принадлежат, по данным Gene Ontology, группам семейства HSP, сигнальных транс дукторов, метабо-лических ферментов, опухолеассоциированных про-теинов и протеинов, контролирующих цитоскелетную перестройку и окислительный стресс. Системный подход к биологическим процессам доказал также, что все эти белки связаны в сети с центральными дви-жущими элементами -c-MYC, p53, РА и ПСА, кото-рые, как известно, являются ключевыми регулятора-ми при возникновении и прогрессии РПЖ и потенциальными мишенями для терапии [12].…”