ALCAM is a Novel Cytoplasmic Membrane Protein in TNF-α Stimulated Invasive Cholangiocarcinoma Cells
| Autor: | Songsak Petmitr, Sopit Wongkham, Polkit Sangvanich, Nantana Suwandittakul, Poom Adisakwattana, Onrapak Reamtong |
|---|---|
| Rok vydání: | 2015 |
| Předmět: |
Fetal Proteins
Proteomics Cytoplasm Cancer Research Epidemiology Cell Adhesion Molecules Neuronal Blotting Western Fluorescent Antibody Technique Apoptosis Real-Time Polymerase Chain Reaction Cholangiocarcinoma Antigen Western blot Antigens CD Cell Movement Tumor Cells Cultured medicine Humans Neoplasm Invasiveness RNA Messenger ALCAM Cell Proliferation biology medicine.diagnostic_test Reverse Transcriptase Polymerase Chain Reaction Tumor Necrosis Factor-alpha Cell growth Cell Membrane Public Health Environmental and Occupational Health Molecular biology Blot Bile Ducts Intrahepatic Bile Duct Neoplasms Oncology Membrane protein Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization biology.protein Antibody |
| Zdroj: | Asian Pacific Journal of Cancer Prevention. 16:3849-3856 |
| ISSN: | 1513-7368 |
| DOI: | 10.7314/apjcp.2015.16.9.3849 |
| Popis: | Background: Cholangiocarcinoma (CCA), or bile duct cancer, is incurable with a high mortality rate due to a lack of effective early diagnosis and treatment. Identifying cytoplasmic membrane proteins of invasive CCA that facilitate cancer progression would contribute toward the development of novel tumor markers and effective chemotherapy. Materials and Methods: An invasive CCA cell line (KKU-100) was stimulated using TNF-α and then biotinylated and purified for mass spectrometry analysis. Novel proteins expressed were selected and their mRNAs expression levels were determined by real-time RT-PCR. In addition, the expression of ALCAM was selected for further observation by Western blot analysis, immunofluorescent imaging, and antibody neutralization assay. Results: After comparing the proteomics profile of TNF-α induced invasive with non-treated control cells, over-expression of seven novel proteins was observed in the cytoplasmic membrane of TNF-α stimulated CCA cells. Among these, ALCAM is a novel candidate which showed significant higher mRNA- and protein levels. Immunofluorescent assay also supported that ALCAM was expressed on the cell membrane of the cancer, with increasing intensity associated with TNF-α. Conclusions: This study indicated that ALCAM may be a novel protein candidate expressed on cytoplasmic membranes of invasive CCA cells that could be used as a biomarker for development of diagnosis, prognosis, and drug or antibody-based targeted therapies in the future. |
| Databáze: | OpenAIRE |
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