iTRAQ Quantitative Proteomic Comparison of Metastatic and Non-Metastatic Uveal Melanoma Tumors.
iTRAQ Quantitative Proteomic Comparison of Metastatic and Non-Metastatic Uveal Melanoma Tumors.
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DOI：
10.1371/journal.pone.0135543
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Singh AD
中科院分区：
文献类型：
--
作者：
Crabb JW;Hu B;Crabb JS;Triozzi P;Saunthararajah Y;Tubbs R;Singh AD
葡萄膜黑色素瘤是成人眼部最常见的恶性肿瘤。总死亡率很高，因为这种侵袭性癌症经常在眼科诊断之前转移。对原发性转移性和非转移性肿瘤进行定量蛋白质组学分析，以深入了解葡萄膜黑色素瘤转移的机制和生物标志物。通过LC MS/MS iTRAQ技术分析了8例转移性和7例非转移性人原发性葡萄膜黑色素瘤肿瘤，其中来自正常死后眼睛的Bruch膜/脉络膜复合体作为对照组织。用iTRAQ标签标记来自肿瘤和对照蛋白的胰蛋白酶肽，通过阳离子交换色谱法进行分级分离，并通过LC MS/MS进行分析。蛋白质鉴定使用Mascot搜索引擎和人Uni-Prot/Swiss-Protein数据库，错误发现≤ 1%;蛋白质定量使用Mascot加权平均法。在5个转移性和5个非转移性肿瘤的训练集中，指定为差异表达的蛋白质表现出定量差异（p ≤ 0.05，t检验）。从训练集开发的逻辑回归模型被用于对五个独立肿瘤的转移状态进行分类。在5例转移性和5例非转移性肿瘤中鉴定和定量的1644种蛋白质中，12种蛋白质在≥ 3例转移性肿瘤中独特发现，28种蛋白质仅在转移性肿瘤中显著升高，30种蛋白质仅在转移性肿瘤中显著降低，31种蛋白质在转移性和非转移性肿瘤之间被指定为差异表达。差异表达的胶原蛋白α-3（VI）和热休克蛋白β-1的Logistic回归模型允许正确预测5个独立肿瘤标本的转移状态。目前的数据为转移性和非转移性葡萄膜黑色素瘤的分子差异提供了新的线索。虽然样本量有限且需要验证，但结果支持胶原蛋白α-3（VI）和热休克蛋白β-1作为葡萄膜黑色素瘤转移的候选生物标志物，并建立葡萄膜黑色素瘤原发性肿瘤的定量蛋白质组学数据库。
Uveal melanoma is the most common malignancy of the adult eye. The overall mortality rate is high because this aggressive cancer often metastasizes before ophthalmic diagnosis. Quantitative proteomic analysis of primary metastasizing and non-metastasizing tumors was pursued for insights into mechanisms and biomarkers of uveal melanoma metastasis. Eight metastatic and 7 non-metastatic human primary uveal melanoma tumors were analyzed by LC MS/MS iTRAQ technology with Bruch’s membrane/choroid complex from normal postmortem eyes as control tissue. Tryptic peptides from tumor and control proteins were labeled with iTRAQ tags, fractionated by cation exchange chromatography, and analyzed by LC MS/MS. Protein identification utilized the Mascot search engine and the human Uni-Prot/Swiss-Protein database with false discovery ≤ 1%; protein quantitation utilized the Mascot weighted average method. Proteins designated differentially expressed exhibited quantitative differences (p ≤ 0.05, t-test) in a training set of five metastatic and five non-metastatic tumors. Logistic regression models developed from the training set were used to classify the metastatic status of five independent tumors. Of 1644 proteins identified and quantified in 5 metastatic and 5 non-metastatic tumors, 12 proteins were found uniquely in ≥ 3 metastatic tumors, 28 were found significantly elevated and 30 significantly decreased only in metastatic tumors, and 31 were designated differentially expressed between metastatic and non-metastatic tumors. Logistic regression modeling of differentially expressed collagen alpha-3(VI) and heat shock protein beta-1 allowed correct prediction of metastasis status for each of five independent tumor specimens. The present data provide new clues to molecular differences in metastatic and non-metastatic uveal melanoma tumors. While sample size is limited and validation required, the results support collagen alpha-3(VI) and heat shock protein beta-1 as candidate biomarkers of uveal melanoma metastasis and establish a quantitative proteomic database for uveal melanoma primary tumors.