Jianzhou Chen1, Bostjan Markelc2, Jakob Kaeppler2, Vivian M L Ogundipe3, Yunhong Cao2, W Gillies McKenna2, Ruth J Muschel4. 1. CRUK/MRC Oxford Institute for Radiation Oncology, University of Oxford, Oxford, United Kingdom; Department of Oncology, University of Oxford, Oxford, United Kingdom; Department of Radiation Oncology, Cancer Hospital of Shantou University Medical College, Shantou, China. 2. CRUK/MRC Oxford Institute for Radiation Oncology, University of Oxford, Oxford, United Kingdom; Department of Oncology, University of Oxford, Oxford, United Kingdom. 3. CRUK/MRC Oxford Institute for Radiation Oncology, University of Oxford, Oxford, United Kingdom; Department of Oncology, University of Oxford, Oxford, United Kingdom; Swammerdam Institute for Life Sciences, University of Amsterdam, Amsterdam, The Netherlands. 4. CRUK/MRC Oxford Institute for Radiation Oncology, University of Oxford, Oxford, United Kingdom; Department of Oncology, University of Oxford, Oxford, United Kingdom. Electronic address: ruth.muschel@oncology.ox.ac.uk.
Abstract
PURPOSE: To investigate the induction of type III interferons (IFNs) in human cancer cells by gamma-rays. METHODS AND MATERIALS: Type III IFN expression in human cancer cell lines after gamma-ray irradiation in vitro was assessed by reverse transcription-quantitative polymerase chain reaction and enzyme-linked immunosorbent assay. Signaling pathways mediating type III IFN induction were examined by a variety of means, including immunoblotting, flow cytometry, confocal imaging, and reverse transcription-quantitative polymerase chain reaction. Key mediators in these pathways were further explored and validated using gene CRISPR knockout or short hairpin RNA knockdown. RESULTS: Exposure to gamma-rays directly induced type III IFNs (mainly IFNL1) in human cancer cell lines in dose- and time-dependent fashions. The induction of IFNL1 was primarily mediated by the cytosolic DNA sensors-STING-TBK1-IRF1 signaling axis, with a lesser contribution from the nuclear factor kappa b signaling in HT29 cells. In addition, type III IFN signaling through its receptors serves as a positive feedback loop, further enhancing IFN expression via up-regulation of the kinases in the STING-TBK1 signaling axis. CONCLUSIONS: Our results suggest that IFNL1 can be up-regulated in human cancer cell lines after gamma-ray treatment. In HT29 cells this induction occurs via the STING pathway, adding another layer of complexity to the understanding of radiation-induced antitumor immunity, and may provide novel insights into IFN-based cancer treatment.
PURPOSE: To investigate the induction of type III interferons (IFNs) in humancancer cells by gamma-rays. METHODS AND MATERIALS: Type III IFN expression in humancancer cell lines after gamma-ray irradiation in vitro was assessed by reverse transcription-quantitative polymerase chain reaction and enzyme-linked immunosorbent assay. Signaling pathways mediating type III IFN induction were examined by a variety of means, including immunoblotting, flow cytometry, confocal imaging, and reverse transcription-quantitative polymerase chain reaction. Key mediators in these pathways were further explored and validated using gene CRISPR knockout or short hairpin RNA knockdown. RESULTS: Exposure to gamma-rays directly induced type III IFNs (mainly IFNL1) in humancancer cell lines in dose- and time-dependent fashions. The induction of IFNL1 was primarily mediated by the cytosolic DNA sensors-STING-TBK1-IRF1 signaling axis, with a lesser contribution from the nuclear factor kappa b signaling in HT29 cells. In addition, type III IFN signaling through its receptors serves as a positive feedback loop, further enhancing IFN expression via up-regulation of the kinases in the STING-TBK1 signaling axis. CONCLUSIONS: Our results suggest that IFNL1 can be up-regulated in humancancer cell lines after gamma-ray treatment. In HT29 cells this induction occurs via the STING pathway, adding another layer of complexity to the understanding of radiation-induced antitumor immunity, and may provide novel insights into IFN-based cancer treatment.
Authors: Ruben S A Goedegebuure; Esther A Kleibeuker; Francesca M Buffa; Kitty C M Castricum; Syed Haider; Iris A Schulkens; Luuk Ten Kroode; Jaap van den Berg; Maarten A J M Jacobs; Anne-Marie van Berkel; Nicole C T van Grieken; Sarah Derks; Ben J Slotman; Henk M W Verheul; Adrian L Harris; Victor L Thijssen Journal: J Exp Clin Cancer Res Date: 2021-05-08
Authors: Edyta Biskup; Brian Daniel Larsen; Leonor Rib; Lasse Folkersen; Omid Niazi; Maria R Kamstrup; Claus Storgaard Sørensen Journal: Cells Date: 2020-11-10 Impact factor: 6.600