Expression of non‑homologous end joining factor, Ku80, is negatively correlated with PD‑L1 expression in cancer cells after X‑ray irradiation

Kumazawa,Takuya; Mori,Yasumasa; Sato,Hiro; Permata,Tiara Bunga Mayang; Uchihara,Yuki; Noda,Shin-Ei; Okada,Kohei; Kakoti,Sangeeta; Suzuki,Keiji; Ikota,Hayato; Yokoo,Hideaki; Gondhowiardjo,Soehartati; Nakano,Takashi; Ohno,Tatsuya; Shibata,Atsushi

doi:10.3892/ol.2021.13147

Expression of non‑homologous end joining factor, Ku80, is negatively correlated with PD‑L1 expression in cancer cells after X‑ray irradiation

Authors:
- Takuya Kumazawa
- Yasumasa Mori
- Hiro Sato
- Tiara Bunga Mayang Permata
- Yuki Uchihara
- Shin-Ei Noda
- Kohei Okada
- Sangeeta Kakoti
- Keiji Suzuki
- Hayato Ikota
- Hideaki Yokoo
- Soehartati Gondhowiardjo
- Takashi Nakano
- Tatsuya Ohno
- Atsushi Shibata
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Affiliations: Department of Radiation Oncology, Graduate School of Medicine, Gunma University, Maebashi, Gunma 3718511, Japan, Department of Radiation Oncology, Faculty of Medicine Universitas Indonesia, Dr Cipto Mangunkusumo National General Hospital, Jakarta 10430, Indonesia, Signal Transduction Program, Gunma University Initiative for Advanced Research, Gunma University, Maebashi, Gunma 3718511, Japan, Department of Radiation Oncology, Comprehensive Cancer Centre, International Medical Centre, Saitama Medical University, Saitama 3501298, Japan, Department of Radiation Medical Sciences, Atomic Bomb Disease Institute, Nagasaki University, Nagasaki 852‑8523, Japan, Clinical Department of Pathology, Gunma University Hospital, Maebashi, Gunma 3718511, Japan, Department of Human Pathology, Gunma University Graduate School of Medicine, Maebashi, Gunma 3718511, Japan, National Institute of Radiological Sciences, National Institute for Quantum and Radiological Science and Technology, Chiba 2638555, Japan
Published online on: November 23, 2021 https://doi.org/10.3892/ol.2021.13147
Article Number: 29
Copyright: © Kumazawa et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The growing importance of antitumour immunity by cancer immunotherapy has prompted studies on radiotherapy‑induced immune response. Previous studies have indicated that programmed cell death‑1 ligand (PD‑L1) expression is regulated by DNA damage signalling. However, PD‑L1 up‑regulation after radiotherapy has not been fully investigated at the clinical level, particularly in the context of expression of DNA repair factors. The present study examined the correlation of mRNA expression between PD‑L1 and non‑homologous end joining (NHEJ) factors using The Cancer Genome Atlas database analysis. Among NHEJ factors, Ku80 mRNA expression was negatively correlated with PD‑L1 mRNA expression levels in several types of cancer (colon adenocarcinoma, breast invasive carcinoma, skin cutaneous melanoma, lung adenocarcinoma, head and neck squamous cell carcinoma, uterine corpus endometrial carcinoma, cervical squamous cell carcinoma and endocervical adenocarcinoma). To verify the negative correlation in clinical samples, the present study analysed whether Ku80 expression levels affected PD‑L1 up‑regulation after radiotherapy using cervical squamous cell carcinoma samples. Quantitative evaluation using software analysis of immunohistochemically stained slides revealed that patients with low Ku80 positivity in biopsy specimens demonstrated increased PD‑L1 expression levels after 10 Gy irradiation (Spearman's rank correlation coefficient=‑0.274; P=0.017). Furthermore, PD‑L1 induction levels in tumour cells after 10 Gy of irradiation were significantly inversely correlated with Ku80 expression levels (Spearman's rank correlation coefficient=‑0.379; P<0.001). The present study also confirmed that short interfering RNA‑mediated Ku80 depletion was associated with greater X‑ray‑induced PD‑L1 up‑regulation in HeLa cells. These results indicated that radiotherapy could enhance PD‑L1 induction in tumour cells with low Ku80 expression in a clinical setting. Furthermore, these data highlighted Ku80 as a potential predictive biomarker for immune checkpoint therapy combined with radiotherapy.

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