同位素编码亲和标签法对乳腺癌血浆蛋白质组的差异分析显示生物胞素酶可作为乳腺癌生物标志物。
Differential profiling of breast cancer plasma proteome by isotope-coded affinity tagging method reveals biotinidase as a breast cancer biomarker.
机构信息
Life Sciences Division, Korea Institute of Science and Technology, Seoul 136-791, Korea.
出版信息
BMC Cancer. 2010 Mar 26;10:114. doi: 10.1186/1471-2407-10-114.
BACKGROUND
Breast cancer is one of the leading causes of women's death worldwide. It is important to discover a reliable biomarker for the detection of breast cancer. Plasma is the most ideal source for cancer biomarker discovery since many cells cross-communicate through the secretion of soluble proteins into blood.
METHODS
Plasma proteomes obtained from 6 breast cancer patients and 6 normal healthy women were analyzed by using the isotope-coded affinity tag (ICAT) labeling approach and tandem mass spectrometry. All the plasma samples used were depleted of highly abundant 6 plasma proteins by immune-affinity column chromatography before ICAT labeling. Several proteins showing differential abundance level were selected based on literature searches and their specificity to the commercially available antibodies, and then verified by immunoblot assays.
RESULTS
A total of 155 proteins were identified and quantified by ICAT method. Among them, 33 proteins showed abundance changes by more than 1.5-fold between the plasmas of breast cancer patients and healthy women. We chose 5 proteins for the follow-up confirmation in the individual plasma samples using immunoblot assay. Four proteins, alpha1-acid glycoprotein 2, monocyte differentiation antigen CD14, biotinidase (BTD), and glutathione peroxidase 3, showed similar abundance ratio to ICAT result. Using a blind set of plasmas obtained from 21 breast cancer patients and 21 normal healthy controls, we confirmed that BTD was significantly down-regulated in breast cancer plasma (Wilcoxon rank-sum test, p = 0.002). BTD levels were lowered in all cancer grades (I-IV) except cancer grade zero. The area under the receiver operating characteristic curve of BTD was 0.78. Estrogen receptor status (p = 0.940) and progesterone receptor status (p = 0.440) were not associated with the plasma BTD levels.
CONCLUSIONS
Our study suggests that BTD is a potential serological biomarker for the detection of breast cancer.
背景
乳腺癌是全球女性死亡的主要原因之一。发现可靠的生物标志物来检测乳腺癌非常重要。由于许多细胞通过将可溶性蛋白分泌到血液中来进行交叉通讯，因此血浆是发现癌症生物标志物的最理想来源。
方法
使用同位素编码亲和标签（ICAT）标记方法和串联质谱法分析了来自 6 名乳腺癌患者和 6 名正常健康女性的血浆蛋白质组。在进行 ICAT 标记之前，所有使用的血浆样本均通过免疫亲和柱色谱法去除高度丰富的 6 种血浆蛋白。根据文献搜索和对市售抗体的特异性，选择了几种显示出差异丰度水平的蛋白质，然后通过免疫印迹分析进行验证。
结果
通过 ICAT 方法共鉴定和定量了 155 种蛋白质。其中，33 种蛋白质在乳腺癌患者和健康女性的血浆之间的丰度变化超过 1.5 倍。我们选择了 5 种蛋白质用于使用免疫印迹测定法在个体血浆样本中进行后续确认。4 种蛋白质，α1-酸性糖蛋白 2、单核细胞分化抗原 CD14、生物素酶（BTD）和谷胱甘肽过氧化物酶 3，与 ICAT 结果显示出相似的丰度比。使用 21 名乳腺癌患者和 21 名正常健康对照者的盲血浆样本进行验证，我们确认 BTD 在乳腺癌血浆中明显下调（Wilcoxon 秩和检验，p = 0.002）。除了癌症零级，所有癌症分级（I-IV）中 BTD 水平均降低。BTD 的接收器工作特征曲线下面积为 0.78。雌激素受体状态（p = 0.940）和孕激素受体状态（p = 0.440）与血浆 BTD 水平无关。
结论
我们的研究表明，BTD 是一种潜在的用于检测乳腺癌的血清生物标志物。
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