Quantitative proteomic analysis to discover potential diagnostic markers and therapeutic targets in human renal cell carcinoma.
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.
Okamura
Noboru
N
Department of Clinical Evaluation of Pharmacotherapy, Kobe University Graduate School of Medicine, Kobe, Japan. okumurak@med.kobe-u.ac.jp
Masuda
Taro
T
Gotoh
Akinobu
A
Shirakawa
Toshiro
T
Terao
Shuji
S
Kaneko
Naoki
N
Suganuma
Kazuki
K
Watanabe
Makoto
M
Matsubara
Toshiya
T
Seto
Ryota
R
Matsumoto
Jun
J
Kawakami
Megumi
M
Yamamori
Motohiro
M
Nakamura
Tsutomu
T
Yagami
Tatsurou
T
Sakaeda
Toshiyuki
T
Fujisawa
Masato
M
Nishimura
Osamu
O
Okumura
Katsuhiko
K
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D014408:Biomarkers, Tumor / Q000032:analysis* / Q000037:antagonists & inhibitors; D015153:Blotting, Western; D002292:Carcinoma, Renal Cell / Q000175:diagnosis / Q000188:drug therapy / Q000378:metabolism*; D005260:Female; D006801:Humans; D007680:Kidney Neoplasms / Q000175:diagnosis / Q000188:drug therapy / Q000378:metabolism*; D008297:Male; D008875:Middle Aged; D020543:Proteome / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D014408:Biomarkers, Tumor; D020543:Proteome