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

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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.

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Abbreviations

LC–MS/MS:

Liquid chromatography and tandem mass spectrometry

DCIS:

Ductal carcinoma in situ

ELISA:

Enzyme-linked immunosorbent assay

HPLC:

High-performance liquid chromatography

SDS–PAGE:

Sodium dodecyl sulfate polyacrylamide gel electrophoresis

IEF:

Isoelectric focusing

2-DE:

Two-dimensional electrophoresis

ROC:

Receiver-operating characteristics

AU:

Arbitrary units

AUC:

Area under the ROC curve

ER:

Estrogen receptor

PgR:

Progesterone receptor

PAI-1:

Plasminogen activator inhibitor-1

uPAR:

Urokinase plasminogen activator receptor

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Acknowledgments

We thank Yoshio Kodera and Mamoru Satoh for their technical assistance and advice for this research.

Conflict of interest

None to declare.

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Authors and Affiliations

  1. Department of General Surgery, Graduate School of Medicine, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba, 260-8670, Japan

    Masami Kadowaki, Takafumi Sangai, Takeshi Nagashima, Masahiro Sakakibara, Hideyuki Yoshitomi, Shigetsugu Takano, Koya Fushimi & Masaru Miyazaki

  2. Department of Molecular Diagnosis, Graduate School of Medicine, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba, 260-8670, Japan

    Masami Kadowaki, Kazuyuki Sogawa, Hiroshi Umemura & Fumio Nomura

  3. Department of Diagnostic Pathology, Graduate School of Medicine, Chiba University, and Department of Pathology, Chiba University Hospital, 1-8-1 Inohana, Chuo-ku, Chiba, 260-8670, Japan

    Yukio Nakatani

Authors
  1. Masami Kadowaki
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  2. Takafumi Sangai
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  3. Takeshi Nagashima
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  4. Masahiro Sakakibara
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  5. Hideyuki Yoshitomi
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  7. Kazuyuki Sogawa
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  9. Koya Fushimi
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  10. Yukio Nakatani
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  11. Fumio Nomura
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  12. Masaru Miyazaki
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Corresponding author

Correspondence to Masami Kadowaki.

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Kadowaki, M., Sangai, T., Nagashima, T. et al. Identification of vitronectin as a novel serum marker for early breast cancer detection using a new proteomic approach. J Cancer Res Clin Oncol 137, 1105–1115 (2011). https://doi.org/10.1007/s00432-010-0974-9

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  • DOI: https://doi.org/10.1007/s00432-010-0974-9

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