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Quantitative proteomic analysis to discover potential diagnostic markers and therapeutic targets in human renal cell carcinoma
Author(s) -
Okamura Noboru,
Masuda Taro,
Gotoh Akinobu,
Shirakawa Toshiro,
Terao Shuji,
Kaneko Naoki,
Suganuma Kazuki,
Watanabe Makoto,
Matsubara Toshiya,
Seto Ryota,
Matsumoto Jun,
Kawakami Megumi,
Yamamori Motohiro,
Nakamura Tsutomu,
Yagami Tatsurou,
Sakaeda Toshiyuki,
Fujisawa Masato,
Nishimura Osamu,
Okumura Katsuhiko
Publication year - 2008
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200700619
Subject(s) - renal cell carcinoma , quantitative proteomics , proteomics , trypsin , blot , carcinoma , cancer research , biology , microbiology and biotechnology , medicine , pathology , biochemistry , gene , enzyme
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.