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[iTRAQ] 小细胞肺癌-糖基化-iTRAQ-Label free

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发表于 2015-12-28 09:51:53 | 显示全部楼层 |阅读模式

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Mol Cell Proteomics. 2014 Jan;13(1):30-48. doi: 10.1074/mcp.M113.028621. Epub 2013 Oct 1.Integrated Glycoproteomics Demonstrates Fucosylated Serum Paraoxonase 1 Alterations in Small Cell Lung Cance

IF=
14年6.564
13年7.254
12年7.251
11年7.398

研究关键词 小细胞肺癌,生物标记物, iTRAQ,label free,糖基化


研究概要
      本研究目的在于通过蛋白质组学技术探索小细胞肺癌的血清糖蛋白生物标记物。 岩藻糖基是最常见的糖基化修饰,糖基化水平的改变是潜在的生物标记物。本研究留取健康人、早期肺癌患者、进展期肺癌患者的血清样本, 富集岩藻糖基化的糖蛋白后,分别利用iTRAQ及Label free技术检测3组样本的蛋白差异,通过生物信息学分析找到4个可能作为生物标记物的差异蛋白后,利用HLE和western blot验证差异蛋白结果,结果发现在患者血清中低表达的蛋白PON1,其岩藻糖基化水平明显增加,它可能可以作为肺癌诊断和预后评估的分子标记物。



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Abstract
Small cell lung cancer (SCLC) is an aggressive type of lung cancer, and the detection of SCLCs at an early stage is necessary for successful therapy and for improving cancer survival rates. Fucosylation is one of the most common glycosylation-based modifications. Increased levels of fucosylation have been reported in a number of pathological conditions, including cancers. In this study, we aimed to identify and validate the aberrant and selective fucosylated glycoproteins in the sera of patients with SCLC. Fucosylated glycoproteins were enriched by the Aleuria aurantia lectin column after serum albumin and IgG depletion. In a narrowed down and comparative data analysis of both label-free proteomics and isobaric peptide-tagging chemistry iTRAQ approaches, the fucosylated glycoproteins were identified as up- or down-regulated in the sera of limited disease and extensive disease stage patients with SCLC. Verification was performed by multiple reaction monitoring-mass spectrometry to select reliable markers. Fourfucosylated proteins, APCS, C9, SERPINA4, and PON1, were selected and subsequently validated by hybrid A. aurantia lectin ELISA (HLE) and Western blotting. Compared with Western blotting, the HLE analysis of these four proteins produced more optimal diagnostic values for SCLC. The PON1 protein levels were significantly reduced in the sera of patients with SCLC, whereas the fucosylation levels of PON1 were significantly increased. Fucosylated PON1 exhibited an area under curve of 0.91 for the extensive disease stage by HLE, whereas the PON1 protein levels produced an area under curve of 0.82 by Western blot. The glycan structural analysis of PON1 by MS/MS identified a biantennary fucosylated glycan modification consisting of a core + 2HexNAc + 1Fuc at increased levels in the sera of patients with SCLC. In addition, the PON1 levels were decreased in the sera of the Lewis lung carcinoma lung cancer mouse model that we examined. Our data suggest that fucosylated protein biomarkers, such as PON1, and their fucosylation levels and patterns can serve as diagnostic and prognostic serological markers for SCLC.


PMID: 24085812
免费全文链接
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3879622/



Mol Cell Proteomics-2014-Ahn-IF7.251.pdf

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发表于 2015-12-28 11:03:02 | 显示全部楼层
嗯嗯 学习一下……
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不明菲行物
不明菲行物  发表于 2015-12-28 11:03:22
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