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| Status |
Public on Jul 08, 2020 |
| Title |
Comparative Analysis of the Development of Acquired Radioresistance in Canine and Human Mammary Cancer Cell Lines |
| Platform organism |
Canis lupus familiaris |
| Sample organisms |
Homo sapiens; Canis lupus familiaris |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
Background: Research using in vitro canine mammary carcinoma cell lines and studies involving naturally occurring canine mammary tumours are not only fundamental models used to advance the understanding of cancer in veterinary patients but are also regarded as excellent translational models for human breast cancer. In human medicine, following disease classification and staging breast cancer is typically treated with multimodal therapies including radiotherapy. 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. Comparative canine and human studies investigating the molecular mechanisms of radioresistance may lead to the development of effective cancer treatments that benefit both species.
Methods: Human (MDA-MB-231, MCF-7 and ZR-751) and canine (REM-134) radioresistant cell lines were established by exposing 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. Intrinsic differences between parental and radioresistant cells were investigated by whole-transcriptome gene expression analysis. Gene enrichment and pathway-focused analyses identified signalling networks differentially activated in radioresistant cells.
Results: Colony formation assays identified a range of intrinsic radiosensitivities in the human and canine parental cell lines with the REM-134 cell line showing significantly greater radioresistance compared to the human cell lines. Colony formation and proliferation assays confirmed radioresistance in all developed radioresistant cell lines. Radioresistant cells exhibited enhanced migration and invasion, with evidence of epithelial-to-mesenchymal-transition. Gene analysis identified that following the acquisition of radioresistance in MCF-7 and ZR-751 cell lines the cell lines 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. MDA-MB-231 and MDA-MB-231 RR cell lines both classified as basal, whereas the REM-134 and REM-134 RR cell lines with enriched for HER2 overexpression. Whole-transcriptome gene expression analysis identified down-regulation of ER signalling genes and up-regulation of genes associated with PI3K, MAPK and WNT pathway activity in radioresistant cell lines derived from ER+ cells; this was confirmed by western blot, which showed increased p-AKT and p-ERK expression following radiation.
Conclusions: To our knowledge, our study is the first to develop a canine radioresistant cell line and provide a comparative genetic and phenotypic analysis of the mechanisms of acquired radioresistance between canine and human cell lines. The cellular process that occur with the development of acquired radioresistance are similar between the human and canine cell lines; this suggests that not only is the canine model an appropriate model to study human disease but that treatment strategies used in human medicine may also be applicable to veterinary patients.
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| Overall design |
Comparative whole-transcriptome gene expression analysis of 3 breast cancer cell lines (MCF-7, ZR-751 and MDA-MB-231) and 1 canine breast cancer cell line (REM-134) and the radio-resistant models derived from these (MCF-7-RR, ZR-751-RR, MDA-MB-231-RR and REM-134-RR, respectively), using Lexogen QuantSeq technology. The study included both radiosensitive and radioresistant models exposed to different levels of radiation therapy (untreated or 2 Gray), with samples collected at different timepoints post-treatment.
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| Contributor(s) |
Gray M, Turnbull AK, Martinez-Perez C |
| Citation(s) |
32851022 |
| |
| Submission date |
May 06, 2020 |
| Last update date |
Aug 31, 2020 |
| Contact name |
Carlos Martinez-Perez |
| E-mail(s) |
[email protected]
|
| Organization name |
The University of Edinburgh
|
| Department |
Institute of Genetics and Molecular Medicine
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| Street address |
Western General Hospital, Crewe Road South
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| City |
Edinburgh |
| ZIP/Postal code |
EH4 2XU |
| Country |
United Kingdom |
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| Platforms (1) |
| GPL28490 |
NextSeq 550 (Canis lupus familiaris) |
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| Samples (10)
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| Relations |
| Reanalysis of |
GSM3415939 |
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GSM3415966 |
| Reanalysis of |
GSM3415967 |
| BioProject |
PRJNA630734 |
| SRA |
SRP260296 |
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