下咽癌相关血浆肿瘤标志物的蛋白质组学筛选 |
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引用本文: | 李军政,田文栋,刘雄,胡水旺,张宝,李湘平.下咽癌相关血浆肿瘤标志物的蛋白质组学筛选[J].南方医科大学学报,2013,33(8):1173. |
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作者姓名: | 李军政 田文栋 刘雄 胡水旺 张宝 李湘平 |
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摘 要: | 目的分析下咽癌患者与健康人血浆中差异表达的蛋白质组,筛选潜在的与下咽癌诊断密切相关的血浆肿瘤标志物。方 法取下咽癌患者与健康人空腹血浆标本各6 例分别等量混合成2 组样本,去除血浆高丰度蛋白后,利用双向电泳(2-DE)进行分 离,经软件分析后找出表达量变化2倍以上的差异蛋白点,接着利用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF/TOF) 进行差异蛋白质鉴定,然后利用Western blotting 对α2-HS-糖蛋白的表达量进行验证。结果下咽癌患者与健康人血浆相比较, 筛选出表达量差异2倍以上的差异蛋白点共11 个,其中表达上调的有5个,表达下调的有6个,进行质谱分析后,表达上调的蛋 白有α2-HS-糖蛋白、结合珠蛋白;表达下调的蛋白有免疫球蛋白κ链C结构域、载脂蛋白A1。Western blotting结果表明,α-2-HS- 糖蛋白的表达上调,与双向电泳结果一致。结论下咽癌患者与健康人血浆的蛋白质表达谱表现出一定差异,这些差异蛋白有可 能成为下咽癌患者早期诊断的特异性血浆肿瘤标志物。
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Screening of plasma tumor biomarkers for hypopharyngeal carcinoma by proteomics method |
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Abstract: | Objective To screen tumor biomarkers in the plasma close related with hypopharyngeal carcinoma. Methods Pooled plasm from 6 patients with hypopharyngeal carcinoma and 6 healthy individuals were collected. After removal of high-abundance plasm proteins, two-dimensional gel electrophoresis (2-DE) was performed to isolate the total proteins, and the protein spots with more than 2-fold differential expressions were detected by 2D analysis software followed by identification by MALDI-TOF/TOF mass spectrometer. Western blotting was performed to validate the expression level of α 2-HS-glycoprotein. Results A total of 11 differentially expressed protein spots were selected, including 5 upregulated proteins and 6 downregulated proteins. MALDI-TOF/TOF identified the upregulated proteins in hypopharyngeal carcinoma patients as alpha-2-HS-glycoprotein and haptoglobin and downregulated ones as Ig kappa chain C region and apolipoprotein A-I. Western blotting demonstrated that α-2-HS- glycoprotein expression level was consistent with that detected by 2-DE. Conclusion Patients with hypopharyngeal carcinoma show different plasma protein profiles from healthy individuals. These differentially expressed proteins may serve as potential specific tumor biomarkers for hypopharyngeal carcinoma.
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