Intracellular calprotectin (S100A8/A9) facilitates DNA damage responses and promotes apoptosis in head and neck squamous cell carcinoma

Prokopios P Argyris; Flávia Saavedra; Chris Malz; Ian A Stone; Yuping Wei; William S Boyle; Karen F Johnstone; Ali Khammanivong; Mark C Herzberg

doi:10.1016/j.oraloncology.2022.106304

Intracellular calprotectin (S100A8/A9) facilitates DNA damage responses and promotes apoptosis in head and neck squamous cell carcinoma

Oral Oncol. 2023 Feb:137:106304. doi: 10.1016/j.oraloncology.2022.106304. Epub 2023 Jan 4.

Authors

Prokopios P Argyris¹, Flávia Saavedra², Chris Malz², Ian A Stone³, Yuping Wei², William S Boyle², Karen F Johnstone², Ali Khammanivong⁴, Mark C Herzberg⁵

Affiliations

¹ Department of Biochemistry, Molecular Biology and Biophysics, University of Minnesota, Minneapolis, MN, USA; Masonic Cancer Center, University of Minnesota, Minneapolis, MN, USA; Institute for Molecular Virology, University of Minnesota, Minneapolis, MN, USA; Center for Genome Engineering, University of Minnesota, Minneapolis, MN, USA; Howard Hughes Medical Institute, University of Minnesota, Minneapolis, MN, USA; Division of Oral and Maxillofacial Pathology, School of Dentistry, University of Minnesota, Minneapolis, MN, USA. Electronic address: argyr005@umn.edu.
² Department of Diagnostic and Biological Sciences, School of Dentistry, University of Minnesota, Minneapolis, MN, USA.
³ Department of Immunology, Microbiology and Virology, School of Medicine and Dentistry, University of Rochester, Rochester, NY, USA.
⁴ Masonic Cancer Center, University of Minnesota, Minneapolis, MN, USA; Department of Veterinary Clinical Sciences, College of Veterinary Medicine, University of Minnesota, St. Paul, MN, USA.
⁵ Department of Diagnostic and Biological Sciences, School of Dentistry, University of Minnesota, Minneapolis, MN, USA. Electronic address: mcherzb@umn.edu.

Abstract

Objectives: In head and neck squamous cell carcinoma (HNSCC), poor prognosis and low survival rates are associated with downregulated calprotectin. Calprotectin (S100A8/A9) inhibits cancer cell migration and invasion and facilitates G2/M cell cycle arrest. We investigated whether S100A8/A9 regulates DNA damage responses (DDR) and apoptosis in HNSCC after chemoradiation.

Materials and methods: Human HNSCC cases in TCGA were analyzed for relationships between S100A8/A9 and expression of apoptosis-related genes. Next, S100A8/A9-expressing and non-expressing carcinoma lines (two different lineages) were exposed to genotoxic agents and assessed for 53BP1 and γH2AX expression and percent of viable/dead cells. Finally, S100A8/A9-wild-type and S100A8/A9^null C57BL/6j mice were treated with 4-NQO to induce oral dysplastic and carcinomatous lesions, which were compared for levels of 53BP1.

Results: In S100A8/A9-high HNSCC tumors, apoptosis-related caspase family member genes were upregulated, whereas genes limiting apoptosis were significantly downregulated based on TCGA analyses. After X-irradiation or camptothecin treatment, S100A8/A9-expressing carcinoma cells (i.e., TR146 and KB-S100A8/A9) showed significantly higher 53BP1 and γH2AX expression, DNA fragmentation, proportions of dead cells, and greater sensitivity to cisplatin than wild-type KB or TR146-S100A8/A9-KD cells. Interestingly, KB-S100A8/A9^Δ113-114 cells showed similar 53BP1 and γH2AX levels to S100A8/A9-negative KB and KB-EGFP cells. After 4-NQO treatment, 53BP1 expression in oral lesions was significantly greater in calprotectin^+/+ than S100A8/A9^null mice.

Conclusions: In HNSCC cells, intracellular calprotectin is strongly suggested to potentiate DDR and promote apoptosis in response to genotoxic agents. Hence, patients with S100A8/A9-high HNSCC may encounter more favorable outcomes because more tumor cells enter apoptosis with increased sensitivity to chemoradiation therapy.

Keywords: 53BP1; Calprotectin; Cisplatin; DNA damage; Double-strand breaks; Head and neck cancer; Head and neck squamous cell carcinoma; Radiation; S100A8/A9; γH2AX.

Publication types

Research Support, N.I.H., Extramural

MeSH terms

Animals
Apoptosis
Calgranulin A / genetics
Calgranulin A / metabolism
Calgranulin B / metabolism
Carcinoma*
Head and Neck Neoplasms* / genetics
Humans
Leukocyte L1 Antigen Complex / metabolism
Mice
Mice, Inbred C57BL
Squamous Cell Carcinoma of Head and Neck

Substances

Calgranulin A
Calgranulin B
Leukocyte L1 Antigen Complex
S100A8 protein, human
S100a8 protein, mouse

Abstract

Publication types

MeSH terms

Substances

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