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Updated: Jul 3, 2026
The Use of Reverse Phase Protein Arrays (RPPA) to Explore Protein Expression Variation within Individual Renal Cell Cancers
Published on: January 22, 2013
Quantitative proteomic analysis to discover potential diagnostic markers and therapeutic targets in human renal cell
Noboru Okamura1, Taro Masuda, Akinobu Gotoh
1Department of Clinical Evaluation of Pharmacotherapy, Kobe University Graduate School of Medicine, Kobe, Japan. okumurak@med.kobe-u.ac.jp
Abstract:
Renal cell carcinoma (RCC) is relatively resistant to chemotherapy and radiotherapy. Recent advances in drug development are providing novel agents for the treatment of RCC, but the effects are still minimal. In addition, there is an urgent need to identify diagnostic markers for RCC. In this report, to discover potential diagnostic markers and therapeutic targets, we subjected RCC samples to a quantitative proteomic analysis utilizing 2-nitrobenzenesulfenyl (NBS) reagent. Proteins were extracted from RCC and adjacent normal tissue, obtained surgically from patients, and labeled with NBS reagent containing six (12)C or (13)C. This was followed by trypsin digestion and the enrichment of labeled peptides. Samples were then subjected to analysis by MALDI-TOF MS. NBS-labeled peptides with a 6 Da difference were identified by MS/MS. Thirty-four proteins were upregulated in more than 60% of the patients of which some were previously known, and some were novel. The identity of a few proteins was confirmed by Western blotting and quantitative real time RT-PCR. The results suggest that NBS-based quantitative proteomic analysis is useful for discovering diagnostic markers and therapeutic targets for RCC.
Insights
Researchers identified new diagnostic markers and therapeutic targets for renal cell carcinoma (RCC) using quantitative proteomics. This novel approach helps find proteins crucial for early RCC detection and treatment strategies.
Area of Science:
- Proteomics
- Oncology
- Biochemistry
Background:
- Renal cell carcinoma (RCC) exhibits resistance to conventional therapies.
- Novel therapeutic agents for RCC show limited efficacy.
- There is a critical need for effective diagnostic markers for RCC.
Purpose of the Study:
- To discover potential diagnostic markers for RCC.
- To identify novel therapeutic targets for RCC treatment.
- To evaluate the utility of 2-nitrobenzenesulfenyl (NBS) reagent-based quantitative proteomic analysis.
Main Methods:
- Quantitative proteomic analysis of RCC and adjacent normal tissues using NBS reagent.
- Labeling of extracted proteins with (12)C or (13)C NBS reagent.
- Peptide enrichment, trypsin digestion, and analysis by MALDI-TOF MS and MS/MS.
Main Results:
- Identified 34 upregulated proteins in over 60% of RCC patients.
- Discovered both known and novel proteins implicated in RCC.
- Confirmed protein identities using Western blotting and qRT-PCR.
Conclusions:
- NBS-based quantitative proteomic analysis is effective for discovering RCC diagnostic markers.
- This proteomic approach can identify potential therapeutic targets for RCC.
- The study highlights promising avenues for advancing RCC diagnostics and therapeutics.
