Journal of Cancer Research and Clinical Oncology

, Volume 137, Issue 7, pp 1105–1115 | Cite as

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

  • Masami Kadowaki
  • Takafumi Sangai
  • Takeshi Nagashima
  • Masahiro Sakakibara
  • Hideyuki Yoshitomi
  • Shigetsugu Takano
  • Kazuyuki Sogawa
  • Hiroshi Umemura
  • Koya Fushimi
  • Yukio Nakatani
  • Fumio Nomura
  • Masaru Miyazaki
Original Paper



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.


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).


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.


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


Breast cancer Proteomics Serum marker Immunodepletion Vitronectin 



Liquid chromatography and tandem mass spectrometry


Ductal carcinoma in situ


Enzyme-linked immunosorbent assay


High-performance liquid chromatography


Sodium dodecyl sulfate polyacrylamide gel electrophoresis


Isoelectric focusing


Two-dimensional electrophoresis


Receiver-operating characteristics


Arbitrary units


Area under the ROC curve


Estrogen receptor


Progesterone receptor


Plasminogen activator inhibitor-1


Urokinase plasminogen activator receptor


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Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Masami Kadowaki
    • 1
    • 2
  • Takafumi Sangai
    • 1
  • Takeshi Nagashima
    • 1
  • Masahiro Sakakibara
    • 1
  • Hideyuki Yoshitomi
    • 1
  • Shigetsugu Takano
    • 1
  • Kazuyuki Sogawa
    • 2
  • Hiroshi Umemura
    • 2
  • Koya Fushimi
    • 1
  • Yukio Nakatani
    • 3
  • Fumio Nomura
    • 2
  • Masaru Miyazaki
    • 1
  1. 1.Department of General Surgery, Graduate School of MedicineChiba UniversityChuo-ku, ChibaJapan
  2. 2.Department of Molecular Diagnosis, Graduate School of MedicineChiba UniversityChuo-ku, ChibaJapan
  3. 3.Department of Diagnostic Pathology, Graduate School of Medicine, Chiba University, and Department of PathologyChiba University HospitalChuo-ku, ChibaJapan

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