title: Quantitative proteomic analysis to discover potential diagnostic markers and therapeutic targets in human renal cell carcinoma.
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.
authors.affiliation: Department of Clinical Evaluation of Pharmacotherapy, Kobe University Graduate School of Medicine, Kobe, Japan. okumurak@med.kobe-u.ac.jp
authors.forename: Noboru
authors.initials: N
authors.lastname: Okamura
authors.forename: Taro
authors.initials: T
authors.lastname: Masuda
authors.forename: Akinobu
authors.initials: A
authors.lastname: Gotoh
authors.forename: Toshiro
authors.initials: T
authors.lastname: Shirakawa
authors.forename: Shuji
authors.initials: S
authors.lastname: Terao
authors.forename: Naoki
authors.initials: N
authors.lastname: Kaneko
authors.forename: Kazuki
authors.initials: K
authors.lastname: Suganuma
authors.forename: Makoto
authors.initials: M
authors.lastname: Watanabe
authors.forename: Toshiya
authors.initials: T
authors.lastname: Matsubara
authors.forename: Ryota
authors.initials: R
authors.lastname: Seto
authors.forename: Jun
authors.initials: J
authors.lastname: Matsumoto
authors.forename: Megumi
authors.initials: M
authors.lastname: Kawakami
authors.forename: Motohiro
authors.initials: M
authors.lastname: Yamamori
authors.forename: Tsutomu
authors.initials: T
authors.lastname: Nakamura
authors.forename: Tatsurou
authors.initials: T
authors.lastname: Yagami
authors.forename: Toshiyuki
authors.initials: T
authors.lastname: Sakaeda
authors.forename: Masato
authors.initials: M
authors.lastname: Fujisawa
authors.forename: Osamu
authors.initials: O
authors.lastname: Nishimura
authors.forename: Katsuhiko
authors.initials: K
authors.lastname: Okumura
mesh_terms: 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
chemical_list: D014408:Biomarkers, Tumor; D020543:Proteome