| 摘要: |
| 目的 探讨乳腺癌患者血清中蛋白的表达变化，筛选并分析乳腺癌诊断的生物学标志物。方法 采用定量蛋白质组学串联质量标签（TMT）标记技术对68例Ⅱ期乳腺癌患者及62例健康女性的血清蛋白进行检测，并进行蛋白质定量分析，筛选乳腺癌患者血清中发生显著变化的差异蛋白。利用UniProt数据库和Proteome Discoverer软件及在线工具STRING对差异蛋白进一步行生物信息学分析，应用蛋白质印迹法和qPCR对变化倍数显著的差异蛋白VIME（上调为健康对照女性的3.918倍）和RAF1（下调为健康对照女性的0.251）进行验证。结果 共获得67种差异蛋白，其中26种表达上调，41种表达下调。基因本体论（GO）注释分析和功能聚类分析显示上述差异蛋白与肿瘤的血管生成、新陈代谢进程、生物黏附能力等相关。蛋白质印迹法和qPCR验证结果显示RAF1蛋白和mRNA在Ⅱ期乳腺癌患者血清中的表达水平均低于健康对照女性，而VIME蛋白和mRNA的表达均高于健康对照女性，与筛选结果一致。结论 定量蛋白质组学TMT法能有效筛选Ⅱ期乳腺癌患者血清中的差异蛋白，其中VIME和RAF1蛋白有望成为乳腺癌淋巴结转移的候选血清标志物。 |
| 关键词: 乳腺肿瘤 蛋白质组学 串联质量标签 生物学肿瘤标志物 |
| DOI：10.16781/j.0258-879x.2017.05.0622 |
| 投稿时间：2017-01-28修订日期：2017-03-31 |
| 基金项目:国家自然科学基金（8150111616）. |
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| TMT-based quantitative proteomics analysis and identification of serum biomarkers of breast cancer |
| LI Xi-zhou,FANG Min,SHENG Yuan,YU En-da* |
(Department of General Surgery, Changhai Hospital, Second Military Medical University, Shanghai 200433, China
*Corresponding author) |
| Abstract: |
| Objective To explore the different expression of serum proteins of the patients with breast cancer and to screen the diagnostic biomarkers. Methods Tandem mass tag (TMT)-based quantitative proteomics technology was used to detect and quantificationally analyze the serum proteins of 68 stage Ⅱ breast cancer patients and 62 healthy females; the differentially expressed proteins were screened. Bioinformatics analysis of the differentially expressed proteins were performed by UniPort Knowledgebase, Proteome Discoverer software and online software STRING (http://www.string-db.org). The protein and mRNA expressions of screened proteins, vimentin (VIME, up-regulated 3.918 folds than that of healthy females) and Raf-1 threonine-protein kinase (RAF1, down-regulated 0.251 folds) were detected by Western blotting and qPCR, respectively. Results We screened and identified 67 differentially expressed proteins, with 26 significantly up-regulated and 41 significantly down-regulated. Gene ontology (GO) and enrichment analysis showed that the differentially expressed proteins were related to tumor angiogenesis, metabolism and bioadhesive capacity. The protein and mRNA expressions of RAF1 in the patients with stage Ⅱ breast cancer were significantly lower than those in healthy females, and the protein and mRNA expressions of VIME were significantly higher, which was similar to the results of screening analysis. Conclusion TMT-based quantitative proteomics can effectively identify the differentially expressed proteins of stage Ⅱ breast cancer. VIME and RAF1 may be potential serum biomarkers of lymph node metastasis of breast cancer. |
| Key words: breast neoplasms proteomics tandem repeat sequences biological tumor markers |
