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Ⅰ期非小细胞肺癌与健康人血清的差异表达蛋白质组学分析

王冬滨,张逊,韩洪利,徐医军,石珍亮   

  1. 天津市胸科医院
  • 收稿日期:2011-12-20 修回日期:2012-03-19 出版日期:2012-10-15 发布日期:2012-10-15
  • 通讯作者: 张逊

Proteomics-based analysis of differentially expressed proteins in the sera of stage I non-small cell lung cancer patients and healthy subjects

  • Received:2011-12-20 Revised:2012-03-19 Published:2012-10-15 Online:2012-10-15
  • Contact: Xun Zhang

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摘要： 目的：寻找与I期非小细胞肺癌（NSCLC）相关的血清差异表达蛋白。方法：收集天津市胸科医院2011年9月—11月经手术、病理证实的6例I期NSCLC患者血清，等量混合后设为肺癌组；同期15例健康体检血清，等量混合后设为健康对照组。利用双向凝胶电泳 (2-DE)和基质辅助激光解吸/电离飞行时间质谱（MALDI-TOF-MS）分离、筛选和鉴定2组间的差异表达蛋白。结果：成功获得了2组的双向凝胶电泳图谱，PDquest软件分析显示，2组间差异大于2倍的蛋白质点共有128个，质谱鉴定后确定了10种蛋白质，其中7种表达量上调，分别为α1-酸性糖蛋白、C1酯酶抑制物、血管紧张素原、转铁蛋白、转甲状腺素蛋白、间-α-胰蛋白酶抑制剂重链H2及无花果酶-3；3种表达量下调，分别为纤连蛋白、补体C4-A及补体C3。结论：应用2-DE及MALDI-TOF-MS在I期NSCLC血清中鉴定出4种与肿瘤关系密切的差异表达蛋白，为开发NSCLC早期诊断标志物提供了重要的候选蛋白。

关键词: 肺肿瘤, 非小细胞性, 电泳, 凝胶, 双向, 基质辅助激光解吸电离飞行时间质谱, 蛋白质组学, 血清

Abstract: Objective: To seek differentially expressed serum proteins related to stage I non-small cell lung cancer (NSCLC). Methods: The sera of lung cancer group were collected and balanced mixed from 6 patients who were surgically and pathologically diagnosed as stage I NSCLC in Tianjin chest hospital from September 2011 to November 2011. Those of control group were collected and balanced mixed from 15 healthy subjects in the corresponding period. Two-dimensional gel electrophoresis (2-DE) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) were employed to screen and identify differentially expressed serum proteins between the two groups. Results: Two-dimensional electrophoresis (2-DE) patterns of serum proteins with high-resolution and reproducibility were successfully obtained from sera of the two groups. The average protein spots were 330±12. One hundred and twenty eight significantly differentially expressed protein spots between the two groups were screened with PDquest software．Ten proteins were successfully identified via peptide mass fingerprinting using MALDI-TOF-MS. Among them, 7 proteins were overexpressed, including alpha-1-acid glycoprotein, C1 inhibitor, angiotensinogen, transferrin, transthyretin, inter-alpha-trypsin inhibitor heavy chain H2 and ficolin-3. Whereas three proteins were underexpressed, including fibronectin, complement C4-A and complement C3. Conclusion: Four differentially expressed proteins close related to cancer can be separated and identified initially in the sera of patients with stage I NSCLC by 2-DE and MALDI-TOF-MS, which may serve as candidate biomarkers for early detection of NSCLC.

Key words: lung neoplasms, non-small cell, electrophoresis, gel, two-dimensional, MALDI-TOF-MS, proteomics, 血清