Comparative Glycoproteomic Profiling of Human Body Fluid between Healthy Controls and Patients with Papillary Thyroid Carcinoma
Comparative Glycoproteomic Profiling of Human Body Fluid between Healthy Controls and Patients with Papillary Thyroid Carcinoma
复制标题
健康对照者和甲状腺乳头状癌患者之间人体体液糖蛋白组学分析的比较。
DOI：
10.1021/acs.jproteome.9b00672
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发表时间：
2020
影响因子：
4.4
通讯作者：
Yang Hao
中科院分区：
文献类型：
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作者：
Zhang Yong;Zhao Wanjun;Zhao Yang;Mao Yonghong;Su Tao;Zhong Yi;Wang Shisheng;Zhai Rui;Cheng Jingqiu;Fang Xiang;Zhu Jingqiang;Yang Hao
甲状腺乳头状癌（PTC）是世界范围内女性最常见的甲状腺癌类型。它主要通过细针穿刺活检（FNAB）来证实，这是一种侵入性诊断方法。负责甲状腺激素生物合成的关键蛋白质是糖基化的。因此，位点特异性糖基化的变化与甲状腺癌相关。PTC患者体液的综合定量蛋白质组学和糖蛋白质组学分析可以识别潜在的非侵入性生物标志物，提高诊断准确性，并阐明肿瘤发展的基本机制。在本研究中，我们展示了一个集成的，高度重复性的，快速的方法，涉及体液蛋白质组和糖蛋白质组分析。此外，该方法可以定量分析蛋白质糖基化。采用亲水相互作用液相色谱法富集健康对照（HC）、PTC和PTC伴桥本甲状腺炎（PHT）患者尿液和血浆中的完整N-糖肽。用三氟乙酸和加热从N-糖肽中除去唾液酸。通过HCD-MS/MS使用阶梯碰撞能量和几个搜索引擎进行定量分析，对去唾液酸-N-糖肽进行分析。从三组的血浆和尿液样品（来自15名受试者的90个样品）中分离出92种改变的蛋白质和134种完整的N-糖肽。据我们所知，这项研究是第一个比较血浆和尿蛋白质组和糖蛋白质组的HC，PTC，PHT。此外，我们揭示了一种新的指标（岩藻糖基化的非岩藻糖基化的N-糖肽或F/NF的比例），通过去唾液酸-N-糖肽分析。这些不同表达的糖蛋白和F/NF可作为有助于临床癌症诊断的生物标志物，并可用于非侵入性地提高诊断准确性。
Papillary thyroid carcinoma (PTC) is the most common type of thyroid cancer among women worldwide. It is confirmed mainly by fine-needle aspiration biopsy (FNAB), an invasive diagnostic method. The key proteins responsible for thyroid hormone biosynthesis are glycosylated. Hence, changes in site-specific glycosylation are associated with thyroid cancer. Integrated quantitative proteomic and glycoproteomic analyses of body fluids from patients with PTC may identify potential noninvasive biomarkers, improve diagnostic accuracy, and elucidate the basic mechanisms of tumor development. In the present study, we demonstrate an integrated, highly-reproducible, rapid method involving body fluid proteome and glycoproteome analysis. Moreover, this method may quantitatively profile protein glycosylation. Intact N-glycopeptides from the urine and plasma of healthy controls (HC), PTC, and PTC with Hashimoto's thyroiditis (PHT) were enriched by hydrophilic interaction liquid chromatography. Sialic acid was removed from the N-glycopeptides with trifluoroacetic acid and heat. The desialo-N-glycopeptides were analyzed by HCD-MS/MS using stepped collision energies and several search engines for quantitative profiling. Ninety-two altered proteins and 134 intact N-glycopeptides were isolated from the plasma and urine samples of the three groups (90 samples from 15 subjects). To the best of our knowledge, this study is the first to compare the plasma and urinary proteomes and glycoproteomes of HC, PTC, and PHT. Moreover, we reveal a novel indicator (ratio of fucosylated to non-fucosylated N-glycopeptide or F/NF) through desialo-N-glycopeptide analysis. These differently expressed glycoproteins and F/NF may serve as biomarkers contributing to clinical cancer diagnostics and could be used to improve diagnostic accuracy noninvasively.