基础医学与临床 ›› 2014, Vol. 34 ›› Issue (8): 1054-1058.

• 研究论文 • 上一篇    下一篇

应用iTRAQ定量蛋白质组学方法筛选乳腺癌患者血清差异表达蛋白

曹美群,孙珂焕,吴安民,陈德珩,吴正治   

  1. 深圳市人民医院
  • 收稿日期:2013-08-09 修回日期:2013-11-25 出版日期:2014-08-05 发布日期:2014-07-15
  • 通讯作者: 吴正治 E-mail:1097838442@qq.com
  • 基金资助:
    广东省自然科学基金项目;广东省科技计划项目

Application of iTRAQ quantitative proteomics in identification of serum biomarkers in breast cancer

  • Received:2013-08-09 Revised:2013-11-25 Online:2014-08-05 Published:2014-07-15

摘要: 目的 筛选乳腺癌患者血清特异性表达蛋白,以寻找乳腺癌早期诊断的生物标记物。方法 采集血清,提取蛋白,iTRAQ标记和ESI-QTOF-QⅡ质谱检测,再搜库和Scaffold软件分析,得到各组间血清差异表达蛋白。进一步采用Western-blotting验证差异蛋白。结果 血清中共鉴定出335个蛋白,达到严格定量标准的蛋白151个;乳腺癌与对照组间共筛选出23个差异表达蛋白;乳腺癌与乳腺纤维瘤组间筛选出8个差异蛋白。Westernblot验证Serotran-sferrin,Vitronectin,soform 1 of Gelsolin 3 个蛋白在各组中的表达结果与iTRAQ检测结果一致。结论 本研究证实iTRAQ-LC-MS/MS 技术能够快速、有效地进行差异蛋白质组学研究,并筛选到多个与乳腺癌发病相关的血清蛋白,有可能作为乳腺癌早期诊断的生物标记。

关键词: 乳腺癌, iTRAQ, 蛋白质组学, 血清, 生物标记物

Abstract: Objective: To explore the presence of informative protein biomarkers in the human serum proteome of breast cancer. Methods:Serum samples were profiled using iTRAQ technology coupled with LC-ESI-QTOF-MS, and Mascot searching. Western-blotting were used for validation of the candidate biomarkers. Results: 335 proteins were identified, and 23 proteins were associated with breast cancer. Three biomarker candidates generated from iTRAQ experiments were successfully verified using Western-blotting . Conclusion: This study provided a global view of potential mechanisms and potentional biomarkers of breast cancer, and demonstrated that iTRAQ combined with LC-ESI-QTOF-MS quantitative proteomics is a powerful tool for biomarker discovery.

Key words: breast cancer, iTRAQ, proteomics, serum, biomarker