Development and characterisation of acquired radioresistant breast cancer cell lines
Abstract Background Radiotherapy plays an important role in the multimodal treatment of breast cancer. The response of a breast tumour to radiation depends not only on its innate radiosensitivity but also on tumour repopulation by cells that have developed radioresistance. Development of effective c...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2019-04-01
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| Series: | Radiation Oncology |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s13014-019-1268-2 |
| _version_ | 1819063663550529536 |
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| author | Mark Gray Arran K. Turnbull Carol Ward James Meehan Carlos Martínez-Pérez Maria Bonello Lisa Y. Pang Simon P. Langdon Ian H. Kunkler Alan Murray David Argyle |
| author_facet | Mark Gray Arran K. Turnbull Carol Ward James Meehan Carlos Martínez-Pérez Maria Bonello Lisa Y. Pang Simon P. Langdon Ian H. Kunkler Alan Murray David Argyle |
| author_sort | Mark Gray |
| collection | DOAJ |
| description | Abstract Background Radiotherapy plays an important role in the multimodal treatment of breast cancer. The response of a breast tumour to radiation depends not only on its innate radiosensitivity but also on tumour repopulation by cells that have developed radioresistance. Development of effective cancer treatments will require further molecular dissection of the processes that contribute to resistance. Methods Radioresistant cell lines were established by exposing MDA-MB-231, MCF-7 and ZR-751 parental cells to increasing weekly doses of radiation. The development of radioresistance was evaluated through proliferation and colony formation assays. Phenotypic characterisation included migration and invasion assays and immunohistochemistry. Transcriptomic data were also generated for preliminary hypothesis generation involving pathway-focused analyses. Results Proliferation and colony formation assays confirmed radioresistance. Radioresistant cells exhibited enhanced migration and invasion, with evidence of epithelial-to-mesenchymal-transition. Significantly, acquisition of radioresistance in MCF-7 and ZR-751 cell lines resulted in a loss of expression of both ERα and PgR and an increase in EGFR expression; based on transcriptomic data they changed subtype classification from their parental luminal A to HER2-overexpressing (MCF-7 RR) and normal-like (ZR-751 RR) subtypes, indicating the extent of phenotypic changes and cellular plasticity involved in this process. Radioresistant cell lines derived from ER+ cells also showed a shift from ER to EGFR signalling pathways with increased MAPK and PI3K activity. Conclusions This is the first study to date that extensively describes the development and characterisation of three novel radioresistant breast cancer cell lines through both genetic and phenotypic analysis. More changes were identified between parental cells and their radioresistant derivatives in the ER+ (MCF-7 and ZR-751) compared with the ER- cell line (MDA-MB-231) model; however, multiple and likely interrelated mechanisms were identified that may contribute to the development of acquired resistance to radiotherapy. |
| first_indexed | 2024-12-21T15:18:15Z |
| format | Article |
| id | doaj.art-f86ee81a892849e88ed5db0ce297194d |
| institution | Directory Open Access Journal |
| issn | 1748-717X |
| language | English |
| last_indexed | 2024-12-21T15:18:15Z |
| publishDate | 2019-04-01 |
| publisher | BMC |
| record_format | Article |
| series | Radiation Oncology |
| spelling | doaj.art-f86ee81a892849e88ed5db0ce297194d2022-12-21T18:59:07ZengBMCRadiation Oncology1748-717X2019-04-0114111910.1186/s13014-019-1268-2Development and characterisation of acquired radioresistant breast cancer cell linesMark Gray0Arran K. Turnbull1Carol Ward2James Meehan3Carlos Martínez-Pérez4Maria Bonello5Lisa Y. Pang6Simon P. Langdon7Ian H. Kunkler8Alan Murray9David Argyle10The Royal (Dick) School of Veterinary Studies and Roslin Institute, University of EdinburghCancer Research UK Edinburgh Centre and Division of Pathology Laboratories, Institute of Genetics and Molecular Medicine, Western General Hospital, University of EdinburghThe Royal (Dick) School of Veterinary Studies and Roslin Institute, University of EdinburghCancer Research UK Edinburgh Centre and Division of Pathology Laboratories, Institute of Genetics and Molecular Medicine, Western General Hospital, University of EdinburghCancer Research UK Edinburgh Centre and Division of Pathology Laboratories, Institute of Genetics and Molecular Medicine, Western General Hospital, University of EdinburghCancer Research UK Edinburgh Centre and Division of Pathology Laboratories, Institute of Genetics and Molecular Medicine, Western General Hospital, University of EdinburghThe Royal (Dick) School of Veterinary Studies and Roslin Institute, University of EdinburghCancer Research UK Edinburgh Centre and Division of Pathology Laboratories, Institute of Genetics and Molecular Medicine, Western General Hospital, University of EdinburghCancer Research UK Edinburgh Centre and Division of Pathology Laboratories, Institute of Genetics and Molecular Medicine, Western General Hospital, University of EdinburghSchool of Engineering, Faraday Building, The King’s Buildings, University of EdinburghThe Royal (Dick) School of Veterinary Studies and Roslin Institute, University of EdinburghAbstract Background Radiotherapy plays an important role in the multimodal treatment of breast cancer. The response of a breast tumour to radiation depends not only on its innate radiosensitivity but also on tumour repopulation by cells that have developed radioresistance. Development of effective cancer treatments will require further molecular dissection of the processes that contribute to resistance. Methods Radioresistant cell lines were established by exposing MDA-MB-231, MCF-7 and ZR-751 parental cells to increasing weekly doses of radiation. The development of radioresistance was evaluated through proliferation and colony formation assays. Phenotypic characterisation included migration and invasion assays and immunohistochemistry. Transcriptomic data were also generated for preliminary hypothesis generation involving pathway-focused analyses. Results Proliferation and colony formation assays confirmed radioresistance. Radioresistant cells exhibited enhanced migration and invasion, with evidence of epithelial-to-mesenchymal-transition. Significantly, acquisition of radioresistance in MCF-7 and ZR-751 cell lines resulted in a loss of expression of both ERα and PgR and an increase in EGFR expression; based on transcriptomic data they changed subtype classification from their parental luminal A to HER2-overexpressing (MCF-7 RR) and normal-like (ZR-751 RR) subtypes, indicating the extent of phenotypic changes and cellular plasticity involved in this process. Radioresistant cell lines derived from ER+ cells also showed a shift from ER to EGFR signalling pathways with increased MAPK and PI3K activity. Conclusions This is the first study to date that extensively describes the development and characterisation of three novel radioresistant breast cancer cell lines through both genetic and phenotypic analysis. More changes were identified between parental cells and their radioresistant derivatives in the ER+ (MCF-7 and ZR-751) compared with the ER- cell line (MDA-MB-231) model; however, multiple and likely interrelated mechanisms were identified that may contribute to the development of acquired resistance to radiotherapy.http://link.springer.com/article/10.1186/s13014-019-1268-2RadioresistanceBreast cancerGlobal gene analysisER and EGFR signallingCharacterisation of radioresistant cell lines |
| spellingShingle | Mark Gray Arran K. Turnbull Carol Ward James Meehan Carlos Martínez-Pérez Maria Bonello Lisa Y. Pang Simon P. Langdon Ian H. Kunkler Alan Murray David Argyle Development and characterisation of acquired radioresistant breast cancer cell lines Radiation Oncology Radioresistance Breast cancer Global gene analysis ER and EGFR signalling Characterisation of radioresistant cell lines |
| title | Development and characterisation of acquired radioresistant breast cancer cell lines |
| title_full | Development and characterisation of acquired radioresistant breast cancer cell lines |
| title_fullStr | Development and characterisation of acquired radioresistant breast cancer cell lines |
| title_full_unstemmed | Development and characterisation of acquired radioresistant breast cancer cell lines |
| title_short | Development and characterisation of acquired radioresistant breast cancer cell lines |
| title_sort | development and characterisation of acquired radioresistant breast cancer cell lines |
| topic | Radioresistance Breast cancer Global gene analysis ER and EGFR signalling Characterisation of radioresistant cell lines |
| url | http://link.springer.com/article/10.1186/s13014-019-1268-2 |
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