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显微切割胰腺癌的蛋白质谱分析及热休克蛋白27作为潜在血清标志物的鉴定。

Protein profiling of microdissected pancreas carcinoma and identification of HSP27 as a potential serum marker.

作者信息

Melle Christian, Ernst Günther, Escher Niko, Hartmann Daniel, Schimmel Bettina, Bleul Annett, Thieme Heike, Kaufmann Roland, Felix Klaus, Friess Helmut M, Settmacher Utz, Hommann Merten, Richter Konrad K, Daffner Wolfgang, Täubig Horst, Manger Thomas, Claussen Uwe, von Eggeling Ferdinand

机构信息

Core Unit Chip Application, Institute of Human Genetics and Anthropology, Medical Faculty at Friedrich Schiller University Jena, Jena, Germany.

出版信息

Clin Chem. 2007 Apr;53(4):629-35. doi: 10.1373/clinchem.2006.079194. Epub 2007 Feb 15.

Abstract

BACKGROUND

Patients with pancreatic adenocarcinomas have a poor prognosis because of late clinical manifestation and the tumor's aggressive nature. We used proteomic techniques to search for markers of pancreatic carcinoma.

METHODS

We performed protein profiling of microdissected cryostat sections of 9 pancreatic adenocarcinomas and 10 healthy pancreatic tissue samples using ProteinChip technology (surface-enhanced laser desorption/ionization). We identified proteins by use of 2-dimensional gel electrophoresis, peptide fingerprint mapping, and immunodepletion and used immunohistochemistry for in situ localization of the proteins found. We used ELISA to quantify these proteins in preoperative serum samples from 35 patients with pancreatic cancer and 37 healthy individuals.

RESULTS

From among the differentially expressed signals that were detected by ProteinChip technology, we identified 2 proteins, DJ-1 and heat shock protein 27 (HSP27). We then detected HSP27 in sera of patients by use of ELISA, indicating a sensitivity of 100% and a specificity of 84% for the recognition of pancreatic cancer.

CONCLUSIONS

The detection of DJ-1 and HSP27 in pure defined tissue and the retrieval of HSP27 in serum by antibody-based methods identifies a potential marker for pancreatic cancer.

摘要

背景

胰腺腺癌患者预后较差,原因是临床表现出现较晚且肿瘤具有侵袭性。我们运用蛋白质组学技术寻找胰腺癌标志物。

方法

我们使用蛋白质芯片技术(表面增强激光解吸/电离)对9例胰腺腺癌和10例健康胰腺组织样本的冷冻切片进行显微切割后的蛋白质谱分析。我们通过二维凝胶电泳、肽指纹图谱和免疫去除法鉴定蛋白质,并使用免疫组织化学对所发现蛋白质进行原位定位。我们使用酶联免疫吸附测定法对35例胰腺癌患者和37例健康个体的术前血清样本中的这些蛋白质进行定量。

结果

从蛋白质芯片技术检测到的差异表达信号中,我们鉴定出2种蛋白质,即DJ-1和热休克蛋白27(HSP27)。然后我们通过酶联免疫吸附测定法在患者血清中检测到HSP27,这表明其对胰腺癌识别的敏感性为100%,特异性为84%。

结论

在纯的特定组织中检测到DJ-1和HSP27,并通过基于抗体的方法在血清中检测到HSP27,确定了一种潜在的胰腺癌标志物。

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