Proteomic Analysis of Membrane-associated Proteins from the Breast Cancer Cell Line MCF7

Cancer Genomics - Proteomics

J. BIGEARD²  R. JOUBERT-CARON²  J. BOUSQUET²  J. HARDOUIN²  L. CANELLE¹  C. PIONNEAU²  M. CARON² 

[1] rotein Biochemistry and Proteomics Laboratory (LBPP), UFR SMBH, Université Paris 13, 93017 Bobigny&D Immunoessais et Protéomique, bioMérieux, Chemin de l'Orme, 69280 Marcy l'Etoile, Francerotein Biochemistry and Proteomics Laboratory (LBPP), UFR SMBH, Université Paris 13, 93017 Bobignyrotein Biochemistry and Proteomics Laboratory (LBPP), UFR SMBH, Université Paris 13, 93017 Bobigny&D Immunoessais et Protéomique, bioMérieux, Chemin de l'Orme, 69280 Marcy l'Etoile, France&D Immunoessais et Protéomique, bioMérieux, Chemin de l'Orme, 69280 Marcy l'Etoile, Francerotein Biochemistry and Proteomics Laboratory (LBPP), UFR SMBH, Université Paris 13, 93017 Bobigny&D Immunoessais et Protéomique, bioMérieux, Chemin de l'Orme, 69280 Marcy l'Etoile, France;rotein Biochemistry and Proteomics Laboratory (LBPP), UFR SMBH, Université Paris 13, 93017 Bobignyrotein Biochemistry and Proteomics Laboratory (LBPP), UFR SMBH, Université Paris 13, 93017 Bobignyrotein Biochemistry and Proteomics Laboratory (LBPP), UFR SMBH, Université Paris 13, 93017 Bobigny

关键词: Biomarker;    proteomics;    membrane proteins;    breast cancer;   

DOI  :  

来源: Delinasios GJ CO

【 摘 要 】

Background: Proteins associated with cancer cell membranes represent targets of choice for humoral immune response as well as potential tumour marker proteins in human malignancies. However, proteomic analysis of these proteins, and more generally of low-soluble proteins, remains difficult. Materials and Methods: The breast cancer cell line MCF7 was selected to evaluate a sequential extraction method that enables simple fractionation of human cell proteins according to their subcellular localization, yielding subproteomes enriched in cytosolic and membrane-associated proteins, respectively. A crude plasma membrane preparation was followed by the solubilisation of proteins using trifluoroethanol (TFE) as co-solvent. Results: Cross-matching and statistical analysis performed for each set of two-dimensional electrophoresis (whole-cell, membrane and soluble extracts) and between the different sets highlighted the reproducibility of the extraction process and its usefulness for proteomic analysis. Eighty-three % of the spots of the gels corresponding to the membrane fraction were not found in the gels of the soluble fraction. Conclusion: Due to its simplicity, the approach described here appears well suited for membrane proteomic investigation of human cancer cells and detection of potential biomarkers undetected by current techniques.

【 授权许可】

Unknown