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Updated: May 25, 2026
The Use of Reverse Phase Protein Arrays (RPPA) to Explore Protein Expression Variation within Individual Renal Cell Cancers
Published on: January 22, 2013
Proteomic analysis in clear cell renal cell carcinoma: identification of differentially expressed protein by 2-D DIGE
Francesca Raimondo1, Claudia Salemi, Clizia Chinello
1Department of Experimental Medicine, Univ. of Milano-Bicocca, Via Cadore 48, 20052 Monza, Italy.
Abstract:
Renal cell carcinoma (RCC), the most common neoplasm affecting the adult kidney, is characterised by heterogeneity of histological subtypes, drug resistance, and absence of molecular markers. Two-dimensional difference gel electrophoresis (2-D DIGE) technology in combination with mass spectrometry (MS) was applied to detect differentially expressed proteins in 20 pairs of RCC tissues and matched adjacent normal kidney cortex (ANK), in order to search for RCC markers. After gel analysis by DeCyder 6.5 and EDA software, differentially expressed protein spots were excised from Deep Purple stained preparative 2DE gel. A total of 100 proteins were identified by MS out of 2500 spots, 23 and 77 of these were, respectively, over- and down-expressed in RCC. The Principal Component Analysis applied to gels and protein spots exactly separated the two sample classes in two groups: RCC and ANK. Moreover, some spots, including ANXA2, PPIA, FABP7 and LEG1, resulted highly differential. The DIGE data were also confirmed by immunoblotting analysis for these proteins. In conclusion, we suggest that applying 2-D DIGE to RCC may provide the basis for a better molecular characterization and for the discovery of candidate biomarkers.
Insights
Researchers identified differentially expressed proteins in renal cell carcinoma (RCC) using 2-D DIGE and mass spectrometry. This study highlights potential new biomarkers for improved molecular characterization of kidney cancer.
Area of Science:
- Proteomics
- Oncology
- Biochemistry
Background:
- Renal cell carcinoma (RCC) presents challenges due to histological diversity, drug resistance, and lack of specific molecular markers.
- Identifying novel biomarkers is crucial for better diagnosis and treatment of kidney cancer.
Purpose of the Study:
- To detect differentially expressed proteins in RCC tissues compared to adjacent normal kidney cortex.
- To identify potential protein biomarkers for renal cell carcinoma.
Main Methods:
- Two-dimensional difference gel electrophoresis (2-D DIGE) coupled with mass spectrometry (MS).
- Analysis of 20 pairs of RCC tissues and matched adjacent normal kidney cortex (ANK).
- Bioinformatic analysis including Principal Component Analysis (PCA) and immunoblotting validation.
Main Results:
- Over 2500 protein spots were analyzed, identifying 100 differentially expressed proteins (23 over-expressed, 77 down-expressed) in RCC.
- Principal Component Analysis effectively distinguished between RCC and ANK samples.
- Specific proteins such as ANXA2, PPIA, FABP7, and LEG1 were identified as highly differential and validated by immunoblotting.
Conclusions:
- 2-D DIGE combined with MS is a powerful approach for molecular characterization of RCC.
- The identified proteins show promise as candidate biomarkers for renal cell carcinoma.
- This research provides a foundation for developing improved diagnostic and therapeutic strategies for kidney cancer.
