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Proteomics ; 5(4): 1160-6, 2005 Mar.
Article in English | MEDLINE | ID: mdl-15759317

ABSTRACT

MG7 is an early gastrointestinal cancer specific monoclonal antibody. It can detect gastric cancer with high sensitivity and specificity. However, the target antigen for MG7 has not been identified. Western blot analysis revealed that the MG7 antibody reproducibly recognized two approximately 35 kDa proteins in the total cell lysates of human gastric carcinoma cell lines KATO III and MKN-45. Using a proteomic approach, we identified these MG7 immunoreactive proteins as the human heterogeneous nuclear ribonucleoprotein A2/B1 (hnRNP A2/B1). Western blot analysis of nuclear and cytosolic fraction of KATO III cells using either MG7 or hnRNP A2/B1 antibodies confirmed that the target antigen is located exclusively in the nucleus. With the use of archival samples, we also found that the level of hnRNP A2/B1 protein was increased in gastric cancer tissues (4 out of 5 patients), when compared to their corresponding matching normal stomach tissue.


Subject(s)
Antibodies, Monoclonal/chemistry , Antibodies, Neoplasm/chemistry , Antigens, Neoplasm/chemistry , Gastrointestinal Neoplasms/metabolism , Gene Expression Regulation, Neoplastic , Heterogeneous-Nuclear Ribonucleoprotein Group A-B/biosynthesis , Heterogeneous-Nuclear Ribonucleoprotein Group A-B/chemistry , Proteomics/methods , Adult , Antibodies, Monoclonal/metabolism , Blotting, Western , Cell Line, Tumor , Cell Nucleus/metabolism , Cytosol/metabolism , Databases, Protein , Electrophoresis, Polyacrylamide Gel , Female , Gastric Mucosa/metabolism , Gastrointestinal Neoplasms/immunology , Humans , Male , Middle Aged , Peptides/chemistry , Polymerase Chain Reaction , Software , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Up-Regulation
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