Proteomic analyses of pancreatic cyst fluids.
Proteomic analyses of pancreatic cyst fluids.
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DOI：
10.1097/mpa.0b013e318193a08f
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发表时间：
2009-03
期刊：
影响因子：
2.9
通讯作者：
Yeung AT
中科院分区：
文献类型：
--
作者：
Ke E;Patel BB;Liu T;Li XM;Haluszka O;Hoffman JP;Ehya H;Young NA;Watson JC;Weinberg DS;Nguyen MT;Cohen SJ;Meropol NJ;Litwin S;Tokar JL;Yeung AT
目前还没有一致可靠、可获得且具有结论性的诊断指标来诊断胰腺囊肿并对其进行风险分层。进行蛋白质组分析以探索胰腺囊肿液，以获得有效的诊断生物标志物。我们前瞻性地招募了 20 名研究参与者，并准备了他们的胰腺囊肿液，专门用于蛋白质组分析。应用的蛋白质组学方法有：1) MALDI-TOF（基质辅助激光解吸电离飞行时间）质谱肽组学与 LC/MS/MS（HPLC-串联质谱）蛋白质鉴定。 2) 二维凝胶电泳。 3) GeLC/MS/MS（胰蛋白酶消化通过 SDS-PAGE 分级并通过 LC/MS/MS 鉴定的蛋白质）。对超过 350 个游离肽的测序表明，肽链外切酶活性使得囊液的肽组学不可靠；囊液中的蛋白酶对蛋白质进行切割，从主要蛋白质中产生数百个蛋白质点，使得 2D 凝胶蛋白质组学难以管理； GeLC/MS/MS 揭示了一组与 CEA（癌胚抗原）相关的潜在生物标志物蛋白。淀粉酶的两种同系物、四种粘蛋白的溶解分子、四种溶解的 CEACAM（CEA 相关细胞粘附分子）和四种 S100 同系物可能是促进未来胰腺囊肿诊断和风险分层的候选生物标志物。这种方法每个样本所需的包囊液少于 40 微升，从而可以分析直径小于 1 厘米的包囊。
There are currently no diagnostic indicators that are consistently reliable, obtainable, and conclusive for diagnosing and risk-stratifying pancreatic cysts. Proteomic analyses were performed to explore pancreatic cyst fluids to yield effective diagnostic biomarkers. We have prospectively recruited 20 research participants and prepared their pancreatic cyst fluids specifically for proteomic analyses. Proteomic approaches applied were: 1) MALDI-TOF (matrix-assisted laser-desorption-ionization time-of-flight) mass spectrometry peptidomics with LC/MS/MS (HPLC-tandem mass spectrometry) protein identification. 2) 2D gel electrophoresis. 3) GeLC/MS/MS (tryptic digestion of proteins fractionated by SDS-PAGE and identified by LC/MS/MS). Sequencing of over 350 free peptides showed that exopeptidase activities rendered peptidomics of cyst fluids unreliable; Protein nicking by proteases in the cyst fluids produced hundreds of protein spots from the major proteins, making 2D gel proteomics unmanageable; GeLC/MS/MS revealed a panel of potential biomarker proteins that correlated with CEA (carcinoembryonic antigen). Two homologs of amylase, solubilized molecules of four mucins, four solubilized CEACAMs (CEA-related cell adhesion molecules), and four S100 homologs, may be candidate biomarkers to facilitate future pancreatic cyst diagnosis and risk-stratification. This approach required less than 40 microliters of cyst fluid per sample, offering the possibility to analyze cysts smaller than 1 cm diameter.