A proteomic approach to investigate potential biomarkers directed against membrane-associated breast cancer proteins.
The identification of specific protein markers for breast cancer would provide the basis for early diagnosis. Particularly, membrane and membrane-associated proteins are rich in targets for antibodies that may constitute suitable biomarkers of carcinogenesis. However, membrane proteins separation using 2-DE remains difficult. In this work, the breast cancer cell line MCF7 was used as source of proteins for the screening of potential cell membrane-associated antigens recognized by autoantibodies in patients with breast cancer and healthy volunteers. The protein extract obtained using trifluoroethanol (TFE) as cosolvent was compared to a total cell lysate protein extract prepared by a current technique. After 2-DE separation of the two extracts, their protein patterns clearly differed. About 63% of the proteins identified in the TFE-extract were predicted to possess at least one transmembrane domain. 2-D blots probed with sera from cancer patients or from healthy volunteers showed that, as expected, additional antigens were provided in the TFE-extract. Thus, the method described here appeared well suited for proteomic investigation of potential biomarkers undetected by current techniques.
Laboratory of Protein Biochemistry and Proteomics, (BioMoCeTi), Paris13 University, Bobigny, France.
Ludovic
L
Canelle
Jordane
J
Bousquet
Cedric
C
Pionneau
Julie
J
Hardouin
Genevieve
G
Choquet-Kastylevsky
Raymonde
R
Joubert-Caron
Michel
M
Caron
D015415:Biomarkers / Q000032:analysis*; D001943:Breast Neoplasms / Q000175:diagnosis / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods; D008565:Membrane Proteins / Q000032:analysis*; D009363:Neoplasm Proteins / Q000032:analysis*; D040901:Proteomics / Q000379:methods*; D014407:Tumor Cells, Cultured
D015415:Biomarkers; D008565:Membrane Proteins; D009363:Neoplasm Proteins