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Identification of proteins of human colorectal carcinoma cell line sw480 by two-dimensional electrophoresis and MALDI-TOF mass spectrometry

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World Journal of Gastroenterology 2005年第30期11卷 4679-4684页

作者： Ying-Tao Zhang Yi-Ping Geng Le Zhou Bao-Chang Lai Lv-Sheng Si Yi-Li Wang The-Key Laboratory of Biomedical Information Engineering of Education Ministry Institute of Cancer Research School of Life Science and Technology Xi'an Jiaotong University Xi'an 710061 Shaanxi Province China Department of Gastroenterology First Hospital Xi'an Jiaotong University Xi'an 710061 Shaanxi Province China

AIM： To conduct bhe proteomic analysis of human colorectal carcinoma cell line, sw480 by using two-dimensional electrophoresis （2-DE） and matrix-assisted laser desorption /ionization-time of flight mass spectrometry （MALDITOFMS）. METHODS： The total proteins of human colorectal carcinoma cell line, sw480 were separated with 2-DE by using immobilized pH gradient strips and visualized by staining with silver nitrate. The gel images were acquired by scanner and 2-DE analysis software, Image Master 2D Elite. Nineteen distinct protein spots were excised from gel randomly and digested in gel by TPCK-trypsin. Mass analysis of the byptic digest peptides mixture was performed by using MALDI-TOF MS. Peptide mass fingerprints （PMFs） obtained by the MALDI-TOF analysis were used to search NCBI, swISS-PROT and MSDB databases by using Mascot software. RESULTS： PMF maps of all spots were obtained by MALDI-TOF MS and thirteen proteins were preliminarily identified. CONCLUSION： The methods of analysis and identification of protein spots of tumor cells in 2-DE gel with silver staining by MALDI-TOF MS derived PMF have been established. Protein expression profile of sw480 has been obtained. It is demonstrated that a combination of proteomics and cell culture is a useful approach to comprehend the process of colon carcinogenesis.

关键词： Colorectal carcinoma sw480 cell line Twodimensional electrophoresis MALDI-TOF MS Peptide mass fingerprinting

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