Original Paper

Journal of Cancer Research and Clinical Oncology

, Volume 137, Issue 7, pp 1105-1115

Identification of vitronectin as a novel serum marker for early breast cancer detection using a new proteomic approach

  • Masami KadowakiAffiliated withDepartment of General Surgery, Graduate School of Medicine, Chiba UniversityDepartment of Molecular Diagnosis, Graduate School of Medicine, Chiba University Email author 
  • , Takafumi SangaiAffiliated withDepartment of General Surgery, Graduate School of Medicine, Chiba University
  • , Takeshi NagashimaAffiliated withDepartment of General Surgery, Graduate School of Medicine, Chiba University
  • , Masahiro SakakibaraAffiliated withDepartment of General Surgery, Graduate School of Medicine, Chiba University
  • , Hideyuki YoshitomiAffiliated withDepartment of General Surgery, Graduate School of Medicine, Chiba University
  • , Shigetsugu TakanoAffiliated withDepartment of General Surgery, Graduate School of Medicine, Chiba University
  • , Kazuyuki SogawaAffiliated withDepartment of Molecular Diagnosis, Graduate School of Medicine, Chiba University
  • , Hiroshi UmemuraAffiliated withDepartment of Molecular Diagnosis, Graduate School of Medicine, Chiba University
  • , Koya FushimiAffiliated withDepartment of General Surgery, Graduate School of Medicine, Chiba University
    • , Yukio NakataniAffiliated withDepartment of Diagnostic Pathology, Graduate School of Medicine, Chiba University, and Department of Pathology, Chiba University Hospital
    • , Fumio NomuraAffiliated withDepartment of Molecular Diagnosis, Graduate School of Medicine, Chiba University
    • , Masaru MiyazakiAffiliated withDepartment of General Surgery, Graduate School of Medicine, Chiba University

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Abstract

Purpose

Breast cancer is the most frequent malignancy in women. However, no useful serum markers with high sensitivity and specificity for the detection of early breast cancer have been identified. The search for biological markers of early breast cancer is of continual interest in experimental and clinical breast cancer research. We recently described a simple and highly reproducible three-step proteome analysis for identifying potential disease-marker candidates among the low-abundance serum proteins.

Methods

Serum samples from breast ductal carcinoma in situ (DCIS) patients and normal controls were subjected to a three-step serum proteome analysis. The steps were the following: first, immunodepletion of most abundant proteins; second, fractionation using reverse-phase high-performance liquid chromatography; and third, separation using two-dimensional electrophoresis (2-DE). Differences revealed by protein staining were further confirmed by Western blotting, immunohistochemical staining, and enzyme-linked immunosorbent assays (ELISA).

Results

Twenty-two upregulated and 26 downregulated spots were detected on the 2-DE gels, and a total of 33 proteins were identified by liquid chromatography and tandem mass spectrometry. Western blotting confirmed that the level of vitronectin was significantly increased in DCIS patients compared with that of normal controls. Immunohistochemical staining of vitronectin in breast cancer tissue revealed high expression in small vessel walls surrounding cancer cells and the extracellular matrix of stroma. Moreover, vitronectin serum concentrations, as measured by ELISA, were significantly increased in patients with DCIS or more advanced breast cancer compared with those of normal controls.

Conclusions

Vitronectin could serve as a promising serum marker for the detection of primary breast cancer.

Keywords

Breast cancer Proteomics Serum marker Immunodepletion Vitronectin