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首页> 外文期刊>BMC Cancer >Proteomic analysis of nipple aspirate fluid from women with early-stage breast cancer using isotope-coded affinity tags and tandem mass spectrometry reveals differential expression of vitamin D binding protein
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Proteomic analysis of nipple aspirate fluid from women with early-stage breast cancer using isotope-coded affinity tags and tandem mass spectrometry reveals differential expression of vitamin D binding protein

机译：使用同位素编码的亲和标签和串联质谱对早期乳腺癌女性的乳头抽吸液进行蛋白质组学分析，揭示了维生素D结合蛋白的差异表达

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Background Isotope-coded affinity tag (ICAT) tandem mass spectrometry (MS) allows for qualitative and quantitative analysis of paired protein samples. We sought to determine whether ICAT technology could quantify and identify differential expression of tumor-specific proteins in nipple aspirate fluid (NAF) from the tumor-bearing and contralateral disease-free breasts of patients with unilateral early-stage breast cancer. Methods Paired NAF samples from 18 women with stage I or II unilateral invasive breast carcinoma and 4 healthy volunteers were analyzed using ICAT labeling, sodium dodecyl sulfate-polyacrylamide gel (SDS-PAGE), liquid chromatography, and MS. Proteins were identified by sequence database analysis. Western blot analysis of NAF from an independent sample set from 12 women (8 with early-stage breast cancer and 4 healthy volunteers) was also performed. Results 353 peptides were identified from tandem mass spectra and matched to peptide sequences in the National Center for Biotechnology Information database. Equal numbers of peptides were up- versus down-regulated. Alpha2HS-glycoprotein [Heavy:Light (H:L) ratio 0.63] was underexpressed in NAF from tumor-bearing breasts, while lipophilin B (H:L ratio 1.42), beta-globin (H:L ratio 1.98), hemopexin (H:L ratio 1.73), and vitamin D-binding protein precursor (H:L ratio 1.82) were overexpressed. Western blot analysis of pooled samples of NAF from healthy volunteers versus NAF from women with breast cancer confirmed the overexpression of vitamin D-binding protein in tumor-bearing breasts. Conclusion ICAT tandem MS was able to identify and quantify differences in specific protein expression between NAF samples from tumor-bearing and disease-free breasts. Proteomic screening techniques using ICAT and NAF may be used to find markers for diagnosis of breast cancer.
机译：背景同位素编码亲和标签（ICAT）串联质谱（MS）可以对成对的蛋白质样品进行定性和定量分析。我们试图确定ICAT技术是否可以量化和鉴定单侧早期乳腺癌患者的带瘤和对侧无病乳房的乳头抽吸液（NAF）中肿瘤特异性蛋白的差异表达。方法采用ICAT标记，十二烷基硫酸钠-聚丙烯酰胺凝胶（SDS-PAGE），液相色谱和质谱法对18例I或II期单侧浸润性乳腺癌女性和4名健康志愿者的配对NAF样品进行分析。通过序列数据库分析鉴定蛋白质。还对来自12名妇女（8名患有早期乳腺癌和4名健康志愿者）的独立样本进行了NAF的蛋白质印迹分析。结果从串联质谱图中鉴定出353种肽，并与美国国家生物技术信息中心数据库中的肽序列匹配。相等数量的肽被上调与下调。在携带肿瘤的乳房中，NAF中Alpha2HS-糖蛋白[重：轻（H：L）比0.63]的表达不足，而亲脂素B（H：L比率1.42），β-珠蛋白（H：L比率1.98），血红素（H ：L比1.73）和维生素D结合蛋白前体（H：L比1.82）过表达。对健康志愿者的NAF样本与乳腺癌女性的NAF样本进行蛋白质印迹分析，证实了荷瘤乳房中维生素D结合蛋白的过表达。结论ICAT串联质谱能够鉴定和定量来自荷瘤和无病乳房的NAF样品之间特异性蛋白质表达的差异。使用ICAT和NAF的蛋白质组学筛选技术可用于寻找诊断乳腺癌的标志物。

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