Identification of vitronectin as a novel serum marker for early breast cancer detection using a new proteomic approach.
OBJECTIVE
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.
Department of General Surgery, Graduate School of Medicine, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba, 260-8670, Japan. masami-kd@nifty.com
Masami
M
Kadowaki
Takafumi
T
Sangai
Takeshi
T
Nagashima
Masahiro
M
Sakakibara
Hideyuki
H
Yoshitomi
Shigetsugu
S
Takano
Kazuyuki
K
Sogawa
Hiroshi
H
Umemura
Koya
K
Fushimi
Yukio
Y
Nakatani
Fumio
F
Nomura
Masaru
M
Miyazaki
D000368:Aged; D014408:Biomarkers, Tumor / Q000097:blood*; D015153:Blotting, Western; D001943:Breast Neoplasms / Q000097:blood* / Q000175:diagnosis; D018270:Carcinoma, Ductal, Breast / Q000097:blood* / Q000175:diagnosis; D002285:Carcinoma, Intraductal, Noninfiltrating / Q000097:blood* / Q000175:diagnosis; D055088:Early Detection of Cancer; D015180:Electrophoresis, Gel, Two-Dimensional; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D008875:Middle Aged; D020543:Proteome / Q000032:analysis*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D019096:Vitronectin / Q000097:blood*
D014408:Biomarkers, Tumor; D020543:Proteome; D019096:Vitronectin
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