Glioblastoma-derived spheroid cultures as an experimental model for analysis of EGFR anomalies
| Autor: | Ewelina Stoczynska-Fidelus, Sylwester Piaskowski, James W. Ironside, Sylwia M. Gresner, Dominika Kulczycka-Wojdala, Pawel P. Liberski, Michał Bieńkowski, Wielisław Papierz, Krystyna Hulas-Bigoszewska, Piotr Rieske, Krzysztof Zakrzewski, Magdalena Zakrzewska, Maciej Kolasa, Monika Witusik-Perkowska, Robert Stawski, Dariusz J. Jaskólski |
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| Jazyk: | angličtina |
| Rok vydání: | 2011 |
| Předmět: |
EGFRvIII
Cellular pathology Cancer Research Time Factors Clinical Neurology Biology Gene dosage Polysomy 7 Adherent Culture Spheroids Cellular Cell Adhesion Tumor Cells Cultured Animals Humans EGFR amplification RNA Messenger Cell Proliferation Regulation of gene expression Brain Neoplasms SOXB1 Transcription Factors Cell Cycle Laboratory Investigation - Human/Animal Tissue Cell cycle Cell cultures Molecular biology In vitro ErbB Receptors Gene Expression Regulation Neoplastic Bromodeoxyuridine Neurology Oncology Cell culture Models Animal Neurology (clinical) Spheroids Glioblastoma |
| Zdroj: | Witusik-Perkowska, M, Rieske, P, Hulas-Bigoszewska, K, Zakrzewska, M, Stawski, R, Kulczycka-Wojdala, D, Bienkowski, M, Stoczynska-Fidelus, E, Gresner, S M, Piaskowski, S, Jaskolski, D J, Papierz, W, Zakrzewski, K, Kolasa, M, Ironside, J W & Liberski, P P 2011, ' Glioblastoma-derived spheroid cultures as an experimental model for analysis of EGFR anomalies ', Journal of Neuro-Oncology, vol. 102, no. 3, pp. 395-407 . https://doi.org/10.1007/s11060-010-0352-0 Journal of Neuro-Oncology |
| DOI: | 10.1007/s11060-010-0352-0 |
| Popis: | Glioblastoma cell cultures in vitro are frequently used for investigations on the biology of tumors or new therapeutic approaches. Recent reports have emphasized the importance of cell culture type for maintenance of tumor original features. Nevertheless, the ability of GBM cells to preserve EGFR overdosage in vitro remains controversial. Our experimental approach was based on quantitative analysis of EGFR gene dosage in vitro both at DNA and mRNA level. Real-time PCR data were verified with a FISH method allowing for a distinction between EGFR amplification and polysomy 7. We demonstrated that EGFR amplification accompanied by EGFRwt overexpression was maintained in spheroids, but these phenomena were gradually lost in adherent culture. We noticed a rapid decrease of EGFR overdosage already at the initial stage of cell culture establishment. In contrast to EGFR amplification, the maintenance of polysomy 7 resulted in EGFR locus gain and stabilization even in long-term adherent culture in serum presence. Surprisingly, the EGFRwt expression pattern did not reflect the latter phenomenon and we observed no overexpression of the tested gene. Moreover, quantitative analysis demonstrated that expression of the truncated variant of receptor-EGFRvIII was preserved in GBM-derived spheroids at a level comparable to the initial tumor tissue. Our findings are especially important in the light of research using glioblastoma culture as the experimental model for testing novel EGFR-targeted therapeutics in vitro, with special emphasis on the most common mutated form of receptor-EGFRvIII. |
| Databáze: | OpenAIRE |
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