Glycoproteomic analysis of WGA-bound glycoprotein biomarkers in sera from patients with lung adenocarcinoma
Glycoproteomic analysis of WGA-bound glycoprotein biomarkers in sera from patients with lung adenocarcinoma
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DOI：
10.1002/elps.200800405
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发表时间：
2009-04-01
期刊：
影响因子：
2.9
通讯作者：
Chen, Shui-Tein
中科院分区：
文献类型：
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作者：
Hongsachart, Piyorot;Huang-Liu, Rosa;Chen, Shui-Tein
肺腺癌患者血清样本中的差异蛋白表达谱可能与癌症发展过程中的糖基化有关。在这项研究中，我们使用了各种糖蛋白质组学的方法来研究不同的糖蛋白质组学的人类正常和肺腺癌血清样本，并调查假定改变糖蛋白生物标志物。在我们的初步筛选中，FITC标记的凝集素染色用于检测特异性糖蛋白谱。麦胚凝集素（WGA）凝集素在两个样品中具有最高水平的与糖蛋白的特异性结合。我们用WGA凝集素亲和法富集血清样品中的糖蛋白，然后用抗触珠蛋白进行免疫共沉淀和2-DE，2-D差异凝胶电泳和MS分析。从这些分析中，我们确定了39个差异表达的蛋白质，包括27个上调蛋白质和12个下调蛋白质。利用生物信息学工具进行蛋白质本体的检索、分类和糖基化位点的预测。此外，三个上调的糖蛋白（脂联素，cerulolasmin和糖基磷脂酰肌醇-80）和两个下调的糖蛋白（细胞周期蛋白H和Fyn）被发现与肺癌的发展进行了验证，通过Western印迹分析。我们认为，这些改变的糖蛋白可能是有用的肺癌的发展和进展的生物标志物。
Differential protein expression profiles in the serum samples from patients with lung adenocarcinoma may be associated with glycosylation during cancer development. In this study, we used various glycoproteomic approaches to investigate the different glycoproteomic profiles of human normal and lung adenocarcinoma serum samples and to investigate putative altered glycoprotein biomarkers. In our preliminary screening, FITC-labeled lectin staining was used for the detection of specific glycoprotein profiles. wheat germ agglutinin (WGA) lectin had the highest level of specific binding to glycoproteins in both samples. We enriched for glycoproteins in the serum samples using WGA lectin affinity and then performed co-immunoprecipitation with anti-haptoglobin and 2-DE, 2-D difference in-gel electrophoresis and MS analyses. From these analyses, we identified 39 differentially expressed proteins, including 27 up-regulated proteins and 12 down-regulated proteins. Bioinformatics tools were used to search for protein ontology, category classifications and prediction of glycosylation sites. In addition, three up-regulated glycoproteins (adiponectin, cerulolasmin and glycosylphosphatidyl-inositol-80) and two downregulated glycoproteins (cyclin H and Fyn) that were found to be correlated with lung cancer development were validated by Western blot analysis. We suggest that these altered glycoproteins may be useful as biomarkers for lung cancer development and progression.