title: Intricate Macrophage-Colorectal Cancer Cell Communication in Response to Radiation.
abstract: Both cancer and tumour-associated host cells are exposed to ionizing radiation when a tumour is subjected to radiotherapy. Macrophages frequently constitute the most abundant tumour-associated immune population, playing a role in tumour progression and response to therapy. The present work aimed to evaluate the importance of macrophage-cancer cell communication in the cellular response to radiation. To address this question, we established monocultures and indirect co-cultures of human monocyte-derived macrophages with RKO or SW1463 colorectal cancer cells, which exhibit higher and lower radiation sensitivity, respectively. Mono- and co-cultures were then irradiated with 5 cumulative doses, in a similar fractionated scheme to that used during cancer patients' treatment (2 Gy/fraction/day). Our results demonstrated that macrophages sensitize RKO to radiation-induced apoptosis, while protecting SW1463 cells. Additionally, the co-culture with macrophages increased the mRNA expression of metabolism- and survival-related genes more in SW1463 than in RKO. The presence of macrophages also upregulated glucose transporter 1 expression in irradiated SW1463, but not in RKO cells. In addition, the influence of cancer cells on the expression of pro- and anti-inflammatory macrophage markers, upon radiation exposure, was also evaluated. In the presence of RKO or SW1463, irradiated macrophages exhibit higher levels of pro-inflammatory TNF, IL6, CCL2 and CCR7, and of anti-inflammatory CCL18. However, RKO cells induce an increase of macrophage pro-inflammatory IL1B, while SW1463 cells promote higher pro-inflammatory CXCL8 and CD80, and also anti-inflammatory VCAN and IL10 levels. Thus, our data demonstrated that macrophages and cancer cells mutually influence their response to radiation. Notably, conditioned medium from irradiated co-cultures increased non-irradiated RKO cell migration and invasion and did not impact on angiogenesis in a chicken embryo chorioallantoic membrane assay. Overall, the establishment of primary human macrophage-cancer cell co-cultures revealed an intricate cell communication in response to ionizing radiation, which should be considered when developing therapies adjuvant to radiotherapy.
authors: i3s-Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Porto, Portugal.|INEB-Institute of Biomedical Engineering, University of Porto, Porto, Portugal.|FEUP-Faculty of Engineering, University of Porto, Porto, Portugal.
authors: Ana T
authors: AT
authors: Pinto
authors: i3s-Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Porto, Portugal.|INEB-Institute of Biomedical Engineering, University of Porto, Porto, Portugal.|ICBAS-Institute of Biomedical Sciences Abel Salazar, University of Porto, Porto, Portugal.
authors: Marta L
authors: ML
authors: Pinto
authors: i3s-Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Porto, Portugal.|IPATIMUP-Institute of Molecular Pathology and Immunology of the University of Porto, Porto, Portugal.
authors: Sérgia
authors: S
authors: Velho
authors: i3s-Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Porto, Portugal.|IPATIMUP-Institute of Molecular Pathology and Immunology of the University of Porto, Porto, Portugal.
authors: Marta T
authors: MT
authors: Pinto
authors: i3s-Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Porto, Portugal.|INEB-Institute of Biomedical Engineering, University of Porto, Porto, Portugal.
authors: Ana P
authors: AP
authors: Cardoso
authors: Radiotherapy Service, Centro Hospitalar S. João, EPE, Porto, Portugal.
authors: Rita
authors: R
authors: Figueira
authors: Radiotherapy Service, Centro Hospitalar S. João, EPE, Porto, Portugal.
authors: Armanda
authors: A
authors: Monteiro
authors: Radiotherapy Service, Centro Hospitalar S. João, EPE, Porto, Portugal.
authors: Margarida
authors: M
authors: Marques
authors: i3s-Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Porto, Portugal.|IPATIMUP-Institute of Molecular Pathology and Immunology of the University of Porto, Porto, Portugal.|Department of Pathology and Oncology, Faculty of Medicine, University of Porto, Porto, Portugal.
authors: Raquel
authors: R
authors: Seruca
authors: i3s-Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Porto, Portugal.|INEB-Institute of Biomedical Engineering, University of Porto, Porto, Portugal.|ICBAS-Institute of Biomedical Sciences Abel Salazar, University of Porto, Porto, Portugal.
authors: Mário A
authors: MA
authors: Barbosa
authors: Department of Radiation Oncology and Experimental Cancer Research, Ghent University Hospital, Ghent, Belgium.
authors: Marc
authors: M
authors: Mareel
authors: i3s-Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Porto, Portugal.|INEB-Institute of Biomedical Engineering, University of Porto, Porto, Portugal.|Department of Pathology and Oncology, Faculty of Medicine, University of Porto, Porto, Portugal.
authors: Maria J
authors: MJ
authors: Oliveira
authors: Centre for Gene Regulation and Expression, College of Life Sciences, University of Dundee, Dundee, United Kingdom.
authors: Sónia
authors: S
authors: Rocha
mesh_terms: D000818:Animals; D002450:Cell Communication / Q000528:radiation effects*; D045744:Cell Line, Tumor; D002642:Chick Embryo; D018920:Coculture Techniques; D015179:Colorectal Neoplasms / Q000378:metabolism / Q000473:pathology* / Q000532:radiotherapy; D051246:Glucose Transport Proteins, Facilitative / Q000378:metabolism; D006801:Humans; D008264:Macrophages / Q000378:metabolism / Q000502:physiology* / Q000528:radiation effects; D009361:Neoplasm Invasiveness
chemical_list: D051246:Glucose Transport Proteins, Facilitative