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Use of quantitative shotgun proteomics to identify fibronectin 1 as a potential plasma biomarker for clear cell carcinoma of the kidney

Article type: Research Article

Authors: Yokomizo, Akiraa; *; 1 | Takakura, Michikob; 1 | Kanai, Yaec | Sakuma, Tomohirod | Matsubara, Junichib | Honda, Kazufumib | Naito, Seijia | Yamada, Tesshib | Ono, Masayab

Affiliations: [a] Department of Urology, Graduate School Medical Sciences, Kyushu University, Fukuoka, Japan | [b] Division of Chemotherapy and Clinical Research, National Cancer Center Research Institute, Tokyo, Japan | [c] Division of Molecular Pathology, National Cancer Center Research Institute, Tokyo, Japan | [d] BioBusiness Group, Mitsui Knowledge Industry, Tokyo, Japan

Correspondence: [*] Corresponding author: Akira Yokomizo, Department of Urology, Graduate School of Medical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8252, Japan. Tel.: +81 92 642 5603; Fax: +81 92 642 5618; E-mail: [email protected].

Note: [1] These two authors contributed equally to this work.

Keywords: Renal cell carcinoma, tumor marker, proteomics, fibronectin

DOI: 10.3233/CBM-2012-0243

Journal: Cancer Biomarkers, vol. 10, no. 3-4, pp. 175-183, 2012

Published: 25 May 2012
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Abstract

Background:

Early detection would be one of the most effective means to improve the outcome of renal cell carcinoma (RCC). We searched for a new plasma marker for RCC using a label-free quantitative shotgun proteomics method.

Methods:

Plasma proteins were digested by trypsin, and the resulting peptides were analyzed by 2-Dimensional Image Converted Analysis of Liquid chromatography mass spectrometry (2DICAL). An identified biomarker candidate was subjected to validation using the Amplified Luminescent Proximity Homogeneous Assay (AlphaLISA).

Results:

Among a total of 23,407 independent MS peaks, we found that the mean intensity of 59 peaks significantly differed between 20 clear cell RCC patients and 20 healthy controls. MS/MS spectra from 16 of the 59 peaks matched the amino acid sequences of the fibronectin 1 (FN1) gene product. The increased plasma level of FN1 in RCC patients was validated in a cohort of in 77 patients and 130 healthy controls (p<0.0001).

Conclusions:

The FN1 is considered to be a promising biomarker candidate for clear cell RCC. Furthermore, AlphaLISA is an alternate to the conventional enzyme-linked immunosorbent assay and should prove useful for the rapid validation of biomarker candidates.

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