Apoptosis-induced ectodomain shedding of hypoxia-regulated carbonic anhydrase IX from tumor cells: a double-edged response to chemotherapy
| Autor: | Silvia Pastorekova, Michal Pastorek, Stanislav Stuchlík, Franck Dequiedt, Jaromir Pastorek, Lenka Jelenska, Olga Sedlakova, Ivana Vidlickova, Miriam Zatovicova |
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| Rok vydání: | 2016 |
| Předmět: |
Male
0301 basic medicine Carbonic anhydrase IX Cancer Research Epithelial-Mesenchymal Transition genetic structures Cell Apoptosis Ectodomain ADAM17 Protein Flow cytometry 03 medical and health sciences Paracrine signalling 0302 clinical medicine Neoplasms Carbonic anhydrase Genetics medicine Chemotherapy Humans Cycloheximide Hypoxia Autocrine signalling Metalloproteinase Shedding biology medicine.diagnostic_test business.industry Antibodies Monoclonal Molecular biology Cell Hypoxia 030104 developmental biology medicine.anatomical_structure Oncology Tumor progression 030220 oncology & carcinogenesis Immunology biology.protein Female business Research Article HeLa Cells |
| Zdroj: | BMC Cancer |
| ISSN: | 1471-2407 |
| DOI: | 10.1186/s12885-016-2267-4 |
| Popis: | Background Carbonic anhydrase IX (CA IX) is a tumor-associated, highly active, transmembrane carbonic anhydrase isoform regulated by hypoxia and implicated in pH control and adhesion-migration-invasion. CA IX ectodomain (ECD) is shed from the tumor cell surface to serum/plasma of patients, where it can signify cancer prognosis. We previously showed that the CA IX ECD release is mediated by disintegrin and metalloproteinase ADAM17. Here we investigated the CA IX ECD shedding in tumor cells undergoing apoptosis in response to cytotoxic drugs, including cycloheximide and doxorubicin. Methods Presence of cell surface CA IX was correlated to the extent of apoptosis by flow cytometry in cell lines with natural or ectopic CA IX expression. CA IX ECD level was assessed by ELISA using CA IX-specific monoclonal antibodies. Effect of recombinant CA IX ECD on the activation of molecular pathways was evaluated using the cell-based dual-luciferase reporter assay. Results We found a significantly lower occurrence of apoptosis in the CA IX-positive cell subpopulation than in the CA IX-negative one. We also demonstrated that the cell-surface CA IX level dropped during the death progress due to an increased ECD shedding, which required a functional ADAM17. Inhibitors of metalloproteinases reduced CA IX ECD shedding, but not apoptosis. The CA IX ECD release induced by cytotoxic drugs was connected to elevated expression of CA IX in the surviving fraction of cells. Moreover, an externally added recombinant CA IX ECD activated a pathway driven by the Nanog transcription factor implicated in epithelial-mesenchymal transition and stemness. Conclusions These findings imply that the increased level of the circulating CA IX ECD might be useful as an indicator of an effective antitumor chemotherapy. Conversely, elevated CA IX ECD might generate unwanted effects through autocrine/paracrine signaling potentially contributing to resistance and tumor progression. |
| Databáze: | OpenAIRE |
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