Identification of proteins of human colorectal carcinoma cell line SW480 by two-dimensional electrophoresis and MALDI-TOF mass spectrometry

• Published in: World journal of gastroenterology : WJG 2005-08, Vol.11 (30), p.4679-4684
• Main Authors: Zhang, Ying-Tao, Geng, Yi-Ping, Zhou, Le, Lai, Bao-Chang, Si, Lv-Sheng, Wang, Yi-Li
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Cell Line, Tumor; Colorectal Cancer; Colorectal Neoplasms - chemistry; Colorectal Neoplasms - genetics; Electrophoresis, Gel, Two-Dimensional; Humans; MALDI-TOF; Molecular Sequence Data; Neoplasm Proteins - chemistry; Neoplasm Proteins - genetics; Neoplasm Proteins - isolation & purification; Peptide Mapping; Proteome - isolation & purification; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; SW480; 二维电泳; 光谱测定法; 结直肠癌; 肿瘤细胞; 蛋白质; 鉴别诊断
• ISSN: 1007-9327; 2219-2840
• DOI: 10.3748/wjg.v11.i30.4679

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.