Qu Li-juan, Ding Yan-qing, Liang Li
Department of Pathology, Southern Medical University, Guangzhou 510515, Chain.
Di Yi Jun Yi Da Xue Xue Bao. 2005 Oct;25(10):1211-5, 1220.
To investigate differential protein expression profiles of human colorectal carcinoma cell lines with different metastatic potentials and screen metastasis-associated proteins for analyzing the relationship between metastasis-associated proteins and the tumorigenesis, progression and metastasis of colorectal carcinomas.
With two-dimensional electrophoresis (2-DE) and matrix assisted laser desorption/ionization-time of flight-mass spectrometry, we analyzed the differentially expressed proteins in two human colorectal carcinoma cell lines SW620 and SW480 with high and low metastatic potentials, and screened for proteins associated with colorectal carcinoma metastasis.
Image analysis software Malenie III demonstrated good match and reproducibility of the 2-DE maps obtained from 3 independent experiments, with 1316+/-62 spots detected for SW620 cells and 1332+/-74 spots for SW480 cells with the average matching rate of 82% and 80%, respectively. The spots distributed in the greatest density at the isoelectric points of 4-7 and relative molecular mass weight of 20,000-70,000. Twenty-five distinctly different protein spots (14 spots for SW620 and 11 for SW480) in-gel digested by TPCK trypsin and 23 peptide mass fingerprint maps were obtained by mass spectrometry. Three highly matched proteins and 14 preliminarily matched proteins or fragments were obtained by analysis with Mascot software in the NCBInr database. Some of these differentially expressed proteins were related to gene transcription, cell cycle, signal transduction, cell apoptosis, etc, with possible involvement of cell differentiation, proliferation, invasion, adhesion, and metastasis of colorectal carcinoma.
The 2-DE protein expression profile of SW620 cells with high metastatic potential displays obvious difference from that of SW480 cells with low metastatic potential, and a variety of proteins can be involved in the metastasis of colorectal carcinoma.
研究具有不同转移潜能的人结肠癌细胞系的差异蛋白质表达谱，筛选与转移相关的蛋白质，以分析转移相关蛋白与结肠癌发生、发展及转移之间的关系。
采用双向电泳（2-DE）和基质辅助激光解吸电离飞行时间质谱技术，分析两种具有高、低转移潜能的人结肠癌细胞系SW620和SW480中差异表达的蛋白质，筛选与结肠癌转移相关的蛋白质。
图像分析软件Malenie III显示，从3次独立实验获得的2-DE图谱匹配良好且具有可重复性，SW620细胞检测到1316±62个点，SW480细胞检测到1332±74个点，平均匹配率分别为82%和80%。这些点在等电点4-7和相对分子质量20,000-70,000处分布密度最大。用TPCK胰蛋白酶对25个明显不同的蛋白质点（SW620为14个点，SW480为11个点）进行胶内消化，通过质谱获得23个肽质量指纹图谱。用Mascot软件在NCBInr数据库中分析得到3个高度匹配的蛋白质以及14个初步匹配的蛋白质或片段。其中一些差异表达的蛋白质与基因转录、细胞周期、信号转导、细胞凋亡等有关，可能参与结肠癌的细胞分化、增殖、侵袭、黏附和转移。
具有高转移潜能的SW620细胞的2-DE蛋白质表达谱与低转移潜能的SW480细胞明显不同，多种蛋白质可能参与结肠癌的转移。
