Stage-related alterations in renal cell carcinoma--comprehensive quantitative analysis by 2D-DIGE and protein network analysis

• Published in: PloS one 2011-07, Vol.6 (7), p.e21867-e21867
• Main Authors: Junker, Heike, Venz, Simone, Zimmermann, Uwe, Thiele, Andrea, Scharf, Christian, Walther, Reinhard
• Format: Article
• Language: English
• Subjects: Adult; Aged; Aged, 80 and over; Biochemistry; Biology; Biomarkers; Biopsy; Blotting, Western; Cancer; Cancer therapies; Carcinoma, Renal Cell - metabolism; Carcinoma, Renal Cell - pathology; Cell cycle; Cluster Analysis; Clustering; Correlation analysis; Cytokines; Cytoskeleton; Diagnostic systems; Endoscopy; Ethics; Evaluation; Female; Fluorescence; Gel electrophoresis; Gene expression; Genomics; Humans; Immunohistochemistry; Kidney cancer; Kidney Neoplasms - metabolism; Kidney Neoplasms - pathology; Kinases; Labeling; Lipid metabolism; Lung cancer; Male; Mass Spectrometry; Medicine; Metabolism; Metastasis; Middle Aged; Mitochondria; Molecular biology; Neoplasia; Neoplasm Proteins - chemistry; Neoplasm Proteins - classification; Neoplasm Proteins - metabolism; Neoplasm Staging; Neoplasms; Network analysis; Otolaryngology; Peptidases; Principal Component Analysis; Principal components analysis; Proteases; Proteins; Proteome - metabolism; Proteomics; Quality control; Quantitative analysis; Renal cell carcinoma; Reproducibility of Results; Robust control; Scientific imaging; Signal Transduction; Spots; Statistical analysis; Statistical methods; Studies; Two dimensional analysis; Two-Dimensional Difference Gel Electrophoresis - methods; Urology; Viral proteins
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0021867

Renal cell carcinoma accounts for about 3% of adult malignancies and 85% of neoplasms arising from the kidney. To identify potential progression markers for kidney cancer we examined non-neoplastic and neoplastic kidney tissue from three groups of patients, which represent different tumor stages (pT1, pT2, pT3) by a fluorescence two-dimensional difference gel electrophoresis (2D-DIGE) approach combined with MALDI-ToF-MS/MS. Delta2D software package was used for gel image based quantification and statistical analysis. Thereby, a comprehensive Principal Component Analysis (PCA) could be performed and allowed a robust quality control of the experiment as well as a classification of the analyzed samples, which correlated with the predicted stages from the pathological examination. Additionally for selected candidate proteins we detected a correlation to the tumor grading as revealed by immunohistochemistry. On the 2D protein map 176 spots out of 989 were detected as at least 2-fold differentially expressed. These spots were analyzed by MALDI-ToF-MS/MS and 187 different proteins were identified. The functional clustering of the identified proteins revealed ten groups. Within these groups we found 86 enzymes, 63 proteins of unknown function, 14 transporter, 8 peptidases and 7 kinases. From the systems biology approach we could map many of these proteins in major pathways involved in remodelling of cytoskeleton, mitochondrial dysfunctions and changes in lipid metabolism. Due to complexity of the highly interconnected pathway network, further expression and functional validation of these proteins might provide new insights in kidney cancer progression to design novel diagnostic and therapeutic strategies.