Evaluation of assays for drug efficacy in a three-dimensional model of the lung
| Autor: | Petra Obexer, Jens M. Kelm, Gabriele Gamerith, Arno Amann, Edith Lorenz, Heinz Zwierzina, Julia M. Huber, Stefan Koeck |
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| Jazyk: | angličtina |
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Cancer Research Lung Neoplasms Computational biology Pharmacology Afatinib Vinblastine Efficacy Tissue Culture Techniques 03 medical and health sciences 0302 clinical medicine Spheroids Cellular medicine Tumor Cells Cultured Humans Lung cancer Lung Cell Proliferation business.industry In vivo mimicking Cell Cycle Hanging-drop Drug testing Vinorelbine General Medicine respiratory system medicine.disease 030104 developmental biology medicine.anatomical_structure Oncology 030220 oncology & carcinogenesis Quinazolines Three-dimensional model Cisplatin Drug Screening Assays Antitumor business Original Article – Cancer Research Three dimensional model |
| Zdroj: | Journal of Cancer Research and Clinical Oncology |
| ISSN: | 1432-1335 0171-5216 |
| Popis: | Background The focus of the outlined work is the establishment of a three-dimensional lung model for various drug-screening applications. Methods The non-small cell lung cancer (NSCLC) cell line Colo699 was cultivated as monolayer (2D) on plates for 5 days or as microtissues (3D) using a hanging-drop system for 5 and 10 days. Cells and microtissues were treated with afatinib (10–80 µM), cisplatin (100–800 µM) or vinorelbine (25–200 µM) for 24 or 48 hours (h). Cell proliferation and viability were analysed by intra-cellular adenosine triphosphate (ATP) and lactate dehydrogenase release (LDH) assays, annexin V/propidium iodide (PI) staining, and cell cycle determination. Microtissue morphology and size, as well as cell death were evaluated via phase contrast microscopy. Results Our results demonstrate the valid determination of viability and cell death using established assays in the 3D system for drug testing. The comparison of ATP, LDH and cytometry data showed moderate (0.40) to very strong (0.99) correlations. Thereby, we observed partially significant differences in drug efficacy between microtissues and 2D cultures dependent from the applied treatment and read-out method. Altogether, microtissues developed resistance to cisplatin and vinorelbine; but remained more vulnerable to afatinib. These findings were confirmed with microscopy. Conclusion In summary, we established an NSCLC 3D test system with multiple assays compatible for drug-testing applications of substances with different mechanisms of action. In addition, our data support the usage of microtissues as more accurate tools for drug-efficacy testing with the possibility of long-term cultivation and treatment. Electronic supplementary material The online version of this article (doi:10.1007/s00432-016-2198-0) contains supplementary material, which is available to authorized users. |
| Databáze: | OpenAIRE |
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