Proteomic Analysis of Urine to Identify Breast Cancer Biomarker Candidates Using a Label-Free LC-MS/MS Approach.
Proteomic Analysis of Urine to Identify Breast Cancer Biomarker Candidates Using a Label-Free LC-MS/MS Approach.
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DOI：
10.1371/journal.pone.0141876
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Li Y
中科院分区：
文献类型：
--
作者：
Beretov J;Wasinger VC;Millar EK;Schwartz P;Graham PH;Li Y
乳腺癌是一种复杂的异质性疾病，是女性死亡的主要原因。早期诊断和监测乳腺癌的进展对改善预后非常重要。本研究的目的是鉴定尿液中的蛋白质生物标志物，用于早期筛查检测和监测浸润性乳腺癌进展。我们使用离子计数相对定量无标记LC-MS/MS分析对来自乳腺癌患者（n = 20）和健康对照女性（n = 20）的尿液进行比较蛋白质组学分析。使用无偏标记的LC-MS/MS蛋白质组学技术提供乳腺癌患者生物系统中丰富蛋白质的谱。数据分析显示，乳腺癌患者与正常对照受试者相比，有59种尿蛋白存在显著差异（p<0.05，倍数变化>3）。36种尿蛋白仅在特定的乳腺癌阶段发现，其中24种增加，12种减少。在鉴定的59种重要的尿蛋白中，发现了13种新的上调蛋白，可用于检测乳腺癌。这些包括与导管原位癌（DCIS）样品中的浸润前乳腺癌（亮氨酸LRC 36、MAST 4和未表征的蛋白质CI 131）、早期浸润性乳腺癌（DYH 8、HBA、PEPA、未表征的蛋白质C4 orf 14（CD 014）、聚丝蛋白和MMRN 2）和转移性乳腺癌（AGRIN、NEGR 1、FIBA和角蛋白KIC 10）相关的阶段特异性标志物。通过蛋白质印迹法在乳腺癌细胞系中对鉴定的3种潜在标志物（ECM 1、MAST 4和聚丝蛋白）进行初步验证。一个潜在的标志物MAST 4在人乳腺癌组织以及个体人乳腺癌尿液样品中分别用免疫组织化学和蛋白质印迹法进一步验证。我们的研究结果表明，尿液是一种有用的非侵入性生物标志物来源，所鉴定的特征模式（生物标志物）具有在BC检测中临床应用的潜力。在以下研究中，需要使用更大的独立患者队列进行验证。
Breast cancer is a complex heterogeneous disease and is a leading cause of death in women. Early diagnosis and monitoring progression of breast cancer are important for improving prognosis. The aim of this study was to identify protein biomarkers in urine for early screening detection and monitoring invasive breast cancer progression. We performed a comparative proteomic analysis using ion count relative quantification label free LC-MS/MS analysis of urine from breast cancer patients (n = 20) and healthy control women (n = 20). Unbiased label free LC-MS/MS-based proteomics was used to provide a profile of abundant proteins in the biological system of breast cancer patients. Data analysis revealed 59 urinary proteins that were significantly different in breast cancer patients compared to the normal control subjects (p<0.05, fold change >3). Thirty-six urinary proteins were exclusively found in specific breast cancer stages, with 24 increasing and 12 decreasing in their abundance. Amongst the 59 significant urinary proteins identified, a list of 13 novel up-regulated proteins were revealed that may be used to detect breast cancer. These include stage specific markers associated with pre-invasive breast cancer in the ductal carcinoma in-situ (DCIS) samples (Leucine LRC36, MAST4 and Uncharacterized protein CI131), early invasive breast cancer (DYH8, HBA, PEPA, uncharacterized protein C4orf14 (CD014), filaggrin and MMRN2) and metastatic breast cancer (AGRIN, NEGR1, FIBA and Keratin KIC10). Preliminary validation of 3 potential markers (ECM1, MAST4 and filaggrin) identified was performed in breast cancer cell lines by Western blotting. One potential marker MAST4 was further validated in human breast cancer tissues as well as individual human breast cancer urine samples with immunohistochemistry and Western blotting, respectively. Our results indicate that urine is a useful non-invasive source of biomarkers and the profile patterns (biomarkers) identified, have potential for clinical use in the detection of BC. Validation with a larger independent cohort of patients is required in the following study.