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首页> 外文期刊>World Journal of Gastroenterology >Loss of clusterin both in serum and tissue correlates with the tumorigenesis of esophageal squamous cell carcinoma via proteomics approaches
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Loss of clusterin both in serum and tissue correlates with the tumorigenesis of esophageal squamous cell carcinoma via proteomics approaches

机译：血清和组织中簇蛋白的丢失与通过蛋白质组学方法研究的食管鳞状细胞癌的发生有关

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AIM: To identify the differentially secreted proteins or polypeptides associated with tumorigenesis of esophageal squamous cell carcinoma (ESCC) from serum and to find potential tumor secreted biomarkers. METHODS: Proteins from human ESCC tissue and its matched adjacent normal tissue; pre-surgery and post-surgery serum; and pre-surgery and normal control serum were separated by two-dimensional electrophoresis (2-DE) to identify differentially expressed proteins. The silver-stained 2-DE were scanned with digital ImageScanner and analyzed with ImageMaster 2D Elite 3.10 software. A cluster of protein spots differentially expressed were selected and identified with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). One of the differentially expressed proteins, clusterin, was down-regulated in cancer tissue and pre-surgery serum, but it was reversed in post-surgery serum. The results were confirmed by semi-quantitative reverse-transcription (RT)-PCR and western blot. RESULTS: Comparisons of the protein spots identified on the 2-DE maps from human matched sera showed that some proteins were differentially expressed, with most of them showing no differences in composition, shape or density. Being analyzed by MALDI-TOF-MS and database searching, clusterin was differentially expressed and down-regulated in both cancer tissue and pre-surgery serum compared with their counterparts. The results were also validated by RT-PCR and western blot. CONCLUSION: The differentially expressed clusterin may play a key role during tumorigenesis of ESCC. The 2DE-MS based proteomic approach is one of the powerful tools for discovery of secreted markers from peripheral.
机译：目的：从血清中鉴定与食管鳞状细胞癌（ESCC）的肿瘤发生有关的差异分泌的蛋白质或多肽，并寻找潜在的肿瘤分泌生物标志物。方法：人ESCC组织及其匹配的正常组织中的蛋白质；手术前和手术后血清；二维电泳（2-DE）分离术前血清和正常对照血清，以鉴定差异表达的蛋白质。用数字ImageScanner扫描银染过的2-DE，并使用ImageMaster 2D Elite 3.10软件进行分析。选择并差异表达的蛋白质斑点簇，并通过基质辅助激光解吸/电离飞行时间质谱（MALDI-TOF-MS）进行鉴定。差异表达的蛋白质之一，簇蛋白，在癌组织和手术前血清中被下调，但在手术后血清中被逆转。半定量逆转录（RT）-PCR和western blot证实了结果。结果：从人匹配血清的2-DE图谱上鉴定出的蛋白质斑点的比较表明，某些蛋白质表达差异，其中大多数蛋白质在组成，形状或密度上没有差异。通过MALDI-TOF-MS和数据库搜索进行分析，与之相比，簇蛋白在癌组织和术前血清中均差异表达并下调。 RT-PCR和蛋白质印迹也验证了结果。结论：簇蛋白差异表达可能在ESCC的发生中起关键作用。基于2DE-MS的蛋白质组学方法是从外围设备中发现分泌标记的强大工具之一。

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