Identification of vitronectin as a novel serum marker for early breast cancer detection using a new proteomic approach
Identification of vitronectin as a novel serum marker for early breast cancer detection using a new proteomic approach
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DOI：
10.1007/s00432-010-0974-9
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发表时间：
2011-07-01
影响因子：
3.6
通讯作者：
Miyazaki, Masaru
中科院分区：
文献类型：
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作者：
Kadowaki, Masami;Sangai, Takafumi;Miyazaki, Masaru
乳腺癌是女性最常见的恶性肿瘤。然而，尚未发现具有高灵敏度和特异性的有用血清标志物来检测早期乳腺癌。寻找早期乳腺癌的生物标志物一直是实验和临床乳腺癌研究的热点。我们最近描述了一种简单且高度可重复的三步蛋白质组分析，用于在低丰度血清蛋白中识别潜在的疾病标志物候选者。对乳腺导管原位癌 (DCIS) 患者和正常对照的血清样本进行了三步血清蛋白质组分析。步骤如下：首先，免疫耗竭最丰富的蛋白质；其次，采用反相高效液相色谱进行分级；第三，使用二维电泳（2-DE）分离。通过蛋白质印迹、免疫组化染色和酶联免疫吸附测定（ELISA）进一步证实了蛋白质染色显示的差异。2-DE凝胶上检测到22个上调点和26个下调点，通过液相色谱和串联质谱鉴定出总共33个蛋白质。 Western blotting证实，与正常对照相比，DCIS患者的玻连蛋白水平显着升高。乳腺癌组织中玻连蛋白的免疫组织化学染色显示癌细胞周围的小血管壁和间质的细胞外基质中高表达。此外，通过 ELISA 测量，与正常对照相比，DCIS 或更晚期乳腺癌患者的玻连蛋白血清浓度显着升高。玻连蛋白可以作为检测原发性乳腺癌的有前途的血清标志物。
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.