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Release of monocyte migration signals by breast cancer cell lines after ablative and fractionated γ-irradiation

BACKGROUND: Radiotherapy, administered in fractionated as well as ablative settings, is an essential treatment component for breast cancer. Besides the direct tumor cell death inducing effects, there is growing evidence that immune mechanisms contribute - at least in part - to its therapeutic succes...

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Autores principales: Hennel, Roman, Brix, Nikko, Seidl, Karin, Ernst, Anne, Scheithauer, Heike, Belka, Claus, Lauber, Kirsten
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3994291/
https://www.ncbi.nlm.nih.gov/pubmed/24666643
http://dx.doi.org/10.1186/1748-717X-9-85
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author Hennel, Roman
Brix, Nikko
Seidl, Karin
Ernst, Anne
Scheithauer, Heike
Belka, Claus
Lauber, Kirsten
author_facet Hennel, Roman
Brix, Nikko
Seidl, Karin
Ernst, Anne
Scheithauer, Heike
Belka, Claus
Lauber, Kirsten
author_sort Hennel, Roman
collection PubMed
description BACKGROUND: Radiotherapy, administered in fractionated as well as ablative settings, is an essential treatment component for breast cancer. Besides the direct tumor cell death inducing effects, there is growing evidence that immune mechanisms contribute - at least in part - to its therapeutic success. The present study was designed to characterize the type and the extent of cell death induced by fractionated and ablative radiotherapy as well as its impact on the release of monocyte migration stimulating factors by dying breast cancer cells. METHODS: Cell death and senescence assays were employed to characterize the response of a panel of breast cancer cell lines with different receptor and p53 status towards γ-irradiation applied in a fractionated (daily doses of 2 Gy) or ablative setting (single dose of 20 Gy). Cell-free culture supernatants were examined for their monocyte migration stimulating potential in transwell migration and 2D chemotaxis/chemokinesis assays. Irradiation-induced transcriptional responses were analyzed by qRT-PCR, and CD39 surface expression was measured by flow cytometry. RESULTS: Fast proliferating, hormone receptor negative breast cancer cell lines with defective p53 predominantly underwent primary necrosis in response to γ-irradiation when applied at a single, ablative dose of 20 Gy, whereas hormone receptor positive, p53 wildtype cells revealed a combination of apoptosis, primary, and secondary (post-apoptotic) necrosis. During necrosis the dying tumor cells released apyrase-sensitive nucleotides, which effectively stimulated monocyte migration and chemokinesis. In hormone receptor positive cells with functional p53 this was hampered by irradiation-induced surface expression of the ectonucleotidase CD39. CONCLUSIONS: Our study shows that ablative radiotherapy potently induces necrosis in fast proliferating, hormone receptor negative breast cancer cell lines with mutant p53, which in turn release monocyte migration and chemokinesis stimulating nucleotides. Future studies have to elucidate, whether these mechanisms might be utilized in order to stimulate intra-tumoral monocyte recruitment and subsequent priming of adaptive anti-tumor immune responses, and which breast cancer subtypes might be best suited for such approaches.
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spelling pubmed-39942912014-04-23 Release of monocyte migration signals by breast cancer cell lines after ablative and fractionated γ-irradiation Hennel, Roman Brix, Nikko Seidl, Karin Ernst, Anne Scheithauer, Heike Belka, Claus Lauber, Kirsten Radiat Oncol Research BACKGROUND: Radiotherapy, administered in fractionated as well as ablative settings, is an essential treatment component for breast cancer. Besides the direct tumor cell death inducing effects, there is growing evidence that immune mechanisms contribute - at least in part - to its therapeutic success. The present study was designed to characterize the type and the extent of cell death induced by fractionated and ablative radiotherapy as well as its impact on the release of monocyte migration stimulating factors by dying breast cancer cells. METHODS: Cell death and senescence assays were employed to characterize the response of a panel of breast cancer cell lines with different receptor and p53 status towards γ-irradiation applied in a fractionated (daily doses of 2 Gy) or ablative setting (single dose of 20 Gy). Cell-free culture supernatants were examined for their monocyte migration stimulating potential in transwell migration and 2D chemotaxis/chemokinesis assays. Irradiation-induced transcriptional responses were analyzed by qRT-PCR, and CD39 surface expression was measured by flow cytometry. RESULTS: Fast proliferating, hormone receptor negative breast cancer cell lines with defective p53 predominantly underwent primary necrosis in response to γ-irradiation when applied at a single, ablative dose of 20 Gy, whereas hormone receptor positive, p53 wildtype cells revealed a combination of apoptosis, primary, and secondary (post-apoptotic) necrosis. During necrosis the dying tumor cells released apyrase-sensitive nucleotides, which effectively stimulated monocyte migration and chemokinesis. In hormone receptor positive cells with functional p53 this was hampered by irradiation-induced surface expression of the ectonucleotidase CD39. CONCLUSIONS: Our study shows that ablative radiotherapy potently induces necrosis in fast proliferating, hormone receptor negative breast cancer cell lines with mutant p53, which in turn release monocyte migration and chemokinesis stimulating nucleotides. Future studies have to elucidate, whether these mechanisms might be utilized in order to stimulate intra-tumoral monocyte recruitment and subsequent priming of adaptive anti-tumor immune responses, and which breast cancer subtypes might be best suited for such approaches. BioMed Central 2014-03-26 /pmc/articles/PMC3994291/ /pubmed/24666643 http://dx.doi.org/10.1186/1748-717X-9-85 Text en Copyright © 2014 Hennel et al.; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research
Hennel, Roman
Brix, Nikko
Seidl, Karin
Ernst, Anne
Scheithauer, Heike
Belka, Claus
Lauber, Kirsten
Release of monocyte migration signals by breast cancer cell lines after ablative and fractionated γ-irradiation
title Release of monocyte migration signals by breast cancer cell lines after ablative and fractionated γ-irradiation
title_full Release of monocyte migration signals by breast cancer cell lines after ablative and fractionated γ-irradiation
title_fullStr Release of monocyte migration signals by breast cancer cell lines after ablative and fractionated γ-irradiation
title_full_unstemmed Release of monocyte migration signals by breast cancer cell lines after ablative and fractionated γ-irradiation
title_short Release of monocyte migration signals by breast cancer cell lines after ablative and fractionated γ-irradiation
title_sort release of monocyte migration signals by breast cancer cell lines after ablative and fractionated γ-irradiation
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3994291/
https://www.ncbi.nlm.nih.gov/pubmed/24666643
http://dx.doi.org/10.1186/1748-717X-9-85
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