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Chemoresistance is mediated by ovarian cancer leader cells in vitro
BACKGROUND: Leader cells are a subset of cancer cells that coordinate the complex cell-cell and cell-matrix interactions required for ovarian cancer migration, invasion, tumour deposition and are negatively associated with progression-free survival and response to therapy. Emerging evidence suggests...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8408956/ https://www.ncbi.nlm.nih.gov/pubmed/34470672 http://dx.doi.org/10.1186/s13046-021-02086-3 |
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author | Karimnia, Nazanin Wilson, Amy L. Green, Emma Matthews, Amelia Jobling, Thomas W. Plebanski, Magdalena Bilandzic, Maree Stephens, Andrew N. |
author_facet | Karimnia, Nazanin Wilson, Amy L. Green, Emma Matthews, Amelia Jobling, Thomas W. Plebanski, Magdalena Bilandzic, Maree Stephens, Andrew N. |
author_sort | Karimnia, Nazanin |
collection | PubMed |
description | BACKGROUND: Leader cells are a subset of cancer cells that coordinate the complex cell-cell and cell-matrix interactions required for ovarian cancer migration, invasion, tumour deposition and are negatively associated with progression-free survival and response to therapy. Emerging evidence suggests leader cells may be enriched in response to chemotherapy, underlying disease recurrence following treatment. METHODS: CRISPR was used to insert a bicistronic T2A-GFP cassette under the native KRT14 (leader cell) promoter. 2D and 3D drug screens were completed in the presence of chemotherapies used in ovarian cancer management. Leader cell; proliferative (Ki67); and apoptotic status (Cleaved Caspase 3) were defined by live cell imaging and flow cytometry. Quantitative real-time PCR defined “stemness” profiles. Proliferation was assessed on the xCELLigence real time cell analyser. Statistical Analysis was performed using unpaired non-parametric t-tests or one-way ANOVA and Tukey’s multiple comparison post hoc. RESULTS: Leader cells represent a transcriptionally plastic subpopulation of ovarian cancer cells that arise independently of cell division or DNA replication, and exhibit a “stemness” profile that does not correlate with epithelial-to-mesenchymal transition. Chemotherapeutics increased apoptosis-resistant leader cells in vitro, who retained motility and expressed known chemo-resistance markers including ALDH1, Twist and CD44v6. Functional impairment of leader cells restored chemosensitivity, with leader cell-deficient lines failing to recover following chemotherapeutic intervention. CONCLUSIONS: Our data demonstrate that ovarian cancer leader cells are resistant to a diverse array of chemotherapeutic agents, and are likely to play a critical role in the recurrence of chemo-resistant disease as drivers of poor treatment outcomes. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13046-021-02086-3. |
format | Online Article Text |
id | pubmed-8408956 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-84089562021-09-01 Chemoresistance is mediated by ovarian cancer leader cells in vitro Karimnia, Nazanin Wilson, Amy L. Green, Emma Matthews, Amelia Jobling, Thomas W. Plebanski, Magdalena Bilandzic, Maree Stephens, Andrew N. J Exp Clin Cancer Res Research BACKGROUND: Leader cells are a subset of cancer cells that coordinate the complex cell-cell and cell-matrix interactions required for ovarian cancer migration, invasion, tumour deposition and are negatively associated with progression-free survival and response to therapy. Emerging evidence suggests leader cells may be enriched in response to chemotherapy, underlying disease recurrence following treatment. METHODS: CRISPR was used to insert a bicistronic T2A-GFP cassette under the native KRT14 (leader cell) promoter. 2D and 3D drug screens were completed in the presence of chemotherapies used in ovarian cancer management. Leader cell; proliferative (Ki67); and apoptotic status (Cleaved Caspase 3) were defined by live cell imaging and flow cytometry. Quantitative real-time PCR defined “stemness” profiles. Proliferation was assessed on the xCELLigence real time cell analyser. Statistical Analysis was performed using unpaired non-parametric t-tests or one-way ANOVA and Tukey’s multiple comparison post hoc. RESULTS: Leader cells represent a transcriptionally plastic subpopulation of ovarian cancer cells that arise independently of cell division or DNA replication, and exhibit a “stemness” profile that does not correlate with epithelial-to-mesenchymal transition. Chemotherapeutics increased apoptosis-resistant leader cells in vitro, who retained motility and expressed known chemo-resistance markers including ALDH1, Twist and CD44v6. Functional impairment of leader cells restored chemosensitivity, with leader cell-deficient lines failing to recover following chemotherapeutic intervention. CONCLUSIONS: Our data demonstrate that ovarian cancer leader cells are resistant to a diverse array of chemotherapeutic agents, and are likely to play a critical role in the recurrence of chemo-resistant disease as drivers of poor treatment outcomes. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13046-021-02086-3. BioMed Central 2021-09-01 /pmc/articles/PMC8408956/ /pubmed/34470672 http://dx.doi.org/10.1186/s13046-021-02086-3 Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data. |
spellingShingle | Research Karimnia, Nazanin Wilson, Amy L. Green, Emma Matthews, Amelia Jobling, Thomas W. Plebanski, Magdalena Bilandzic, Maree Stephens, Andrew N. Chemoresistance is mediated by ovarian cancer leader cells in vitro |
title | Chemoresistance is mediated by ovarian cancer leader cells in vitro |
title_full | Chemoresistance is mediated by ovarian cancer leader cells in vitro |
title_fullStr | Chemoresistance is mediated by ovarian cancer leader cells in vitro |
title_full_unstemmed | Chemoresistance is mediated by ovarian cancer leader cells in vitro |
title_short | Chemoresistance is mediated by ovarian cancer leader cells in vitro |
title_sort | chemoresistance is mediated by ovarian cancer leader cells in vitro |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8408956/ https://www.ncbi.nlm.nih.gov/pubmed/34470672 http://dx.doi.org/10.1186/s13046-021-02086-3 |
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