1317 PROTEOMIC ANALYSIS OF LASER-MICRODISSECTED FORMALIN-FIXED AND PARAFFIN-EMBEDDED TISSUES OF LOW AND HIGH GLEASON SCORE AND METASTATIC PROSTATE CANCER

Abstract

You have accessJournal of UrologyProstate Cancer: Basic Research II1 Apr 20121317 PROTEOMIC ANALYSIS OF LASER-MICRODISSECTED FORMALIN-FIXED AND PARAFFIN-EMBEDDED TISSUES OF LOW AND HIGH GLEASON SCORE AND METASTATIC PROSTATE CANCER Takahiro Kimura, Toshihiro Yamamoto, Makoto Kihara, Yuko Kamata, Naoki Tanaka, Makiko Otsuji, Yasuhiko Bando, Sayaka Mikami, Bungo Furusato, Hiroyuki Takahashi, Toshihide Nishimura, and Shin Egawa Takahiro KimuraTakahiro Kimura Tokyo, Japan More articles by this author , Toshihiro YamamotoToshihiro Yamamoto Tokyo, Japan More articles by this author , Makoto KiharaMakoto Kihara Kanagawa, Japan More articles by this author , Yuko KamataYuko Kamata Tokyo, Japan More articles by this author , Naoki TanakaNaoki Tanaka Kanagawa, Japan More articles by this author , Makiko OtsujiMakiko Otsuji Kanagawa, Japan More articles by this author , Yasuhiko BandoYasuhiko Bando Tokyo, Japan More articles by this author , Sayaka MikamiSayaka Mikami Tokyo, Japan More articles by this author , Bungo FurusatoBungo Furusato Tokyo, Japan More articles by this author , Hiroyuki TakahashiHiroyuki Takahashi Tokyo, Japan More articles by this author , Toshihide NishimuraToshihide Nishimura Kanagawa, Japan More articles by this author , and Shin EgawaShin Egawa Tokyo, Japan More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2012.02.1664AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES New biomarkers of prostate cancer (PCa) which distinguish between aggressive cancers with metastatic potential and cancers with low risk for progression are required. Although proteomic analysis is useful for discovery of cancer biomarkers, PCa has some difficulties. Collecting fresh and homogeneous cancer tissue from prostatectomy (RP) specimens is difficult because PCa is multifocal and heterogeneous and accuracy of pathological diagnosis of frozen section is limited. In present study, we performed shotgun LC/MS-based global proteomics using recently developed technique, laser-microdissection (LMD) of formalin-fixed and paraffin-embedded (FFPE) tissues to identify grade/stage related biomarkers from low and high Gleason score (GS) and metastatic PCa. METHODS The nanoLC-MS/MS proteomics was performed using FFPE tissues obtained from RP specimens with low GS, high GS and biopsy specimens with metastatic cancer (n=5). Normal epithelium from RP specimens were used as control. Targeted cells were microdissected and proteins were extracted, digested and subjected to LC/MS. Identified proteins were semi-quantified by spectral counting and subjected further to statistical analysis. Candidate proteins were extracted from 3D-scatter plot analysis based on G-test. Extracted candidates were validated using immunohistochemistry (IHC) in 92 RP specimens. RESULTS Total 371 proteins were identified. According to 3D-scatter plot analysis, we have extracted 11 proteins as low GS specific marker candidates and 23 proteins as high GS specific marker candidates. Validation analysis using IHC revealed that methylcrotonoyl-CoA carboxylase b (MCCC2) and fatty acid synthase (FASN) were significantly higher expressed in cancer cells comparing to normal cells (p>0.0001 and p>0.001, respectively). CONCLUSIONS We established the global clinical shotgun proteomics that could work excellently within the small patients number feasibility study in PCa. To achieve such performance, combination of LMD of FFPE specimens, high-resolution mass spectrometry, spectral counting method and 3D-scatter plot analysis are indispensable. MCCC2 and FASN were suggested as biomarkers with different expression profiles associated with grade/stage of PCa. © 2012 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 187Issue 4SApril 2012Page: e534 Advertisement Copyright & Permissions© 2012 by American Urological Association Education and Research, Inc.MetricsAuthor Information Takahiro Kimura Tokyo, Japan More articles by this author Toshihiro Yamamoto Tokyo, Japan More articles by this author Makoto Kihara Kanagawa, Japan More articles by this author Yuko Kamata Tokyo, Japan More articles by this author Naoki Tanaka Kanagawa, Japan More articles by this author Makiko Otsuji Kanagawa, Japan More articles by this author Yasuhiko Bando Tokyo, Japan More articles by this author Sayaka Mikami Tokyo, Japan More articles by this author Bungo Furusato Tokyo, Japan More articles by this author Hiroyuki Takahashi Tokyo, Japan More articles by this author Toshihide Nishimura Kanagawa, Japan More articles by this author Shin Egawa Tokyo, Japan More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...