人胰腺导管腺癌组织及癌旁组织双向电泳图谱的差异分析

目的:探讨人胰腺导管腺癌组织和癌旁组织之间差异的表达蛋白。方法:利用双向凝胶电泳（2-DE）对8例胰腺导管腺癌组织和癌旁组织的总蛋白进行分离,并用质谱仪对两组间差异蛋白质点进行肽指纹谱和串联质谱鉴定。利用蛋白质印迹分析和免疫组化法检测差异蛋白在胰腺癌和癌旁组织的表达。结果:2-DE显示,肿瘤组织中有28个蛋白质点表达上调,17个表达下调。上述蛋白质点经质谱鉴定得到30个蛋白质,包括酶类、抗氧化蛋白、信号转导蛋白、钙结合蛋白、结构蛋白及分子伴侣等。Western blot和免疫组化结果显示差异表达蛋白annexin II在胰腺癌组织中表达上调,与2-DE结果一致。结论:以2-DE为基础的蛋白质组学技术是研究肿瘤的一种重要手段。本实验所得的annexin II等差异表达蛋白可能成为潜在的诊断胰腺癌的分子标志或控制肿瘤生长的治疗靶点。

Differential analysis of two-dimensional gel electrophoresis profile of pancreatic ductal adenocarcinoma and adjacent tissue

Objective To study the difference of expression of proteins in pancreatic cancer and adjacent fissue.Methods Proteomes of eight pairs of pancreatic ductal adenocarcinoma tissue samples and adjacent tissue samples were obtained by high resolution two-dimensional gel electrophoresis(2-DE).Comprehensive analyses of proteins were focused on total protein spots exhibiting statistical alternations between the two groups.Protein identification was done by peptide mass fingerprinting with tandem mass spectrometry(MS/MS).In addition,Western blotting and immunohistochemistry were performed to verify the expression of certain candidate protein.Results A total of 28 protein spot-features were found to be significantly increased and 17 significantly decreased in tumor tissues.Thirty of these protein spots were identified,which included enzymes,antioxidant proteins,signal transduction proteins,calcium-binding protein,structural proteins,chaperones and others.Western blotting and IHC further validated up-regulated expressions of one candidate protein(annexin II)in tumor tissues.Conclusions The analysis of proteomics with 2-DE on human tissue is a useful method for discovering valuable cancer marker candidates.These differential expressed proteins may serve as biomarkers for early detection and therapeutic targets to pancreatic cancer.