两种大肠癌转移性细胞株双向电泳图谱的初步分析  

Analysis of protein expression maps of two metastatic colorectal carcinoma cell lines generated by two-dimensional gel electrophoresis

作　　者：梁莉[1] 丁彦青[1] 李欣[1] 江培洲[1] 李雪华[1] 张进华[1] 

机构地区：[1]南方医科大学病理教研室,广东广州510515

出　　处：《第一军医大学学报》2005年第1期18-21,共4页Journal of First Military Medical University

基　　金：国家重点研究发展规划973项目(2001C13510208);国家自然科学基金(30170423);军队医药卫生基金(01MA128)

摘　　要：目的初步分析大肠癌转移过程中差异表达的蛋白质,为阐述大肠癌转移的分子机制提供线索。方法以一对来自同一亲本、转移能力不同的人大肠癌细胞株SW620和SW480为实验对象,利用双向电泳技术(two-dimensionalgelelectrophoresis,2-DE),建立并优化两种细胞株的蛋白表达图谱;之后利用Melanieш软件分析与大肠癌转移相关的差异表达蛋白点。结果两种细胞株的2-DE蛋白表达谱具有良好的重复性和可比性;采用了非线性pH3-10及重叠窄范围的固相pH梯度胶条,提高了2-DE图谱的分辨率;初步选择11个差异表达蛋白点。结论非线性及重叠窄范围的固相pH梯度胶条可改善2-DE中蛋白分离的效果,初步分离的差异蛋白点为进一步鉴定大肠癌转移相关蛋白及建立大肠癌转移性细胞株的2-DE数据库奠定了基础。Objective To analyze the differentially expressed proteins in metastatic colorectal carcinoma and provide clues for the molecular mechanisms of colorectal cancer metastasis. Methods A pair of colorectal carcinoma cell lines SW620 and SW480 with different metastatic potentials from the same parent cell lines were studied. The protein expression maps of the two cell lines were obtained and optimized using two-dimensional gel electrophoresis (2-DE), and the differentially expressed protein spots associated with metastasis were analyzed by Melanie ш software. Results The 2-DE expression maps of the two cell lines were well matched. The resolution of 2-DE was increased after using the nonlinear pH3-10 or overlapping narrow immobilized pH gradient strips, and 11 differentially expressed protein spots were identified after analysis using the software. Conclusions Nonlinear pH3-10 and overlapping narrow immobilized pH gradient strips have higher separation efficiency for protein in 2-DE. The differentially expressed proteins separated will be useful for identifying metastasis-associated proteins of colorectal carcinoma and for establishing 2-DE database of the metastatic colorectal carcinoma cell lines.

关 键 词：转移 大肠癌 双向电泳 非线性梯度 重叠窄范围的固相pH梯度胶条 

分 类 号：R735.34[医药卫生—肿瘤]