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

Prokopios Argyris; Flávia Saavedra; Christopher R 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

Prokopios Argyris, Flávia Saavedra, Christopher R Malz, Ian A Stone, Yuping Wei, William S. Boyle, Karen F. Johnstone, Ali Khammanivong, Mark C. Herzberg

Research output: Contribution to journal › Article › peer-review

5 Scopus citations

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.

Original language	English (US)
Article number	106304
Journal	Oral Oncology
Volume	137
DOIs	https://doi.org/10.1016/j.oraloncology.2022.106304
State	Published - Feb 2023

Bibliographical note

Funding Information:
The project was supported by NIH/NIDCR R01DE021206 to MCH and R90DE023058 to PPA. The content is solely the responsibility of the authors and does not necessarily represent the official views of the NIDCR or the NIH. The authors thank Dr. Shorouk Elnagdy for assistance with the mice in the 4-NQO protocol, Drs. Rajaram Gopalakrishnan and Ioannis G. Koutlas, University of Minnesota, for assistance with mouse tissue processing for histopathologic analysis, and Dr. Gay Herzberg for expert editing.

Funding Information:
The project was supported by NIH/NIDCR R01DE021206 to MCH and R90DE023058 to PPA. The content is solely the responsibility of the authors and does not necessarily represent the official views of the NIDCR or the NIH. The authors thank Dr. Shorouk Elnagdy for assistance with the mice in the 4-NQO protocol, Drs. Rajaram Gopalakrishnan and Ioannis G. Koutlas, University of Minnesota, for assistance with mouse tissue processing for histopathologic analysis, and Dr. Gay Herzberg for expert editing.

Publisher Copyright:
© 2022

Keywords

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

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1016/j.oraloncology.2022.106304

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@article{3d1788a8b85a4a869cfba9288e8f9c71,

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

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/A9null 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/A9Δ113-114 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/A9null 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",

author = "Prokopios Argyris and Fl{\'a}via Saavedra and Malz, {Christopher R} and Stone, {Ian A} and Yuping Wei and Boyle, {William S.} and Johnstone, {Karen F.} and Ali Khammanivong and Herzberg, {Mark C.}",

note = "Funding Information: The project was supported by NIH/NIDCR R01DE021206 to MCH and R90DE023058 to PPA. The content is solely the responsibility of the authors and does not necessarily represent the official views of the NIDCR or the NIH. The authors thank Dr. Shorouk Elnagdy for assistance with the mice in the 4-NQO protocol, Drs. Rajaram Gopalakrishnan and Ioannis G. Koutlas, University of Minnesota, for assistance with mouse tissue processing for histopathologic analysis, and Dr. Gay Herzberg for expert editing. Funding Information: The project was supported by NIH/NIDCR R01DE021206 to MCH and R90DE023058 to PPA. The content is solely the responsibility of the authors and does not necessarily represent the official views of the NIDCR or the NIH. The authors thank Dr. Shorouk Elnagdy for assistance with the mice in the 4-NQO protocol, Drs. Rajaram Gopalakrishnan and Ioannis G. Koutlas, University of Minnesota, for assistance with mouse tissue processing for histopathologic analysis, and Dr. Gay Herzberg for expert editing. Publisher Copyright: {\textcopyright} 2022",

year = "2023",

month = feb,

doi = "10.1016/j.oraloncology.2022.106304",

language = "English (US)",

volume = "137",

journal = "Oral Oncology",

issn = "1368-8375",

publisher = "Elsevier Ltd",

}

TY - JOUR

T1 - Intracellular calprotectin (S100A8/A9) facilitates DNA damage responses and promotes apoptosis in head and neck squamous cell carcinoma

AU - Argyris, Prokopios

AU - Saavedra, Flávia

AU - Malz, Christopher R

AU - Stone, Ian A

AU - Wei, Yuping

AU - Boyle, William S.

AU - Johnstone, Karen F.

AU - Khammanivong, Ali

AU - Herzberg, Mark C.

N1 - Funding Information: The project was supported by NIH/NIDCR R01DE021206 to MCH and R90DE023058 to PPA. The content is solely the responsibility of the authors and does not necessarily represent the official views of the NIDCR or the NIH. The authors thank Dr. Shorouk Elnagdy for assistance with the mice in the 4-NQO protocol, Drs. Rajaram Gopalakrishnan and Ioannis G. Koutlas, University of Minnesota, for assistance with mouse tissue processing for histopathologic analysis, and Dr. Gay Herzberg for expert editing. Funding Information: The project was supported by NIH/NIDCR R01DE021206 to MCH and R90DE023058 to PPA. The content is solely the responsibility of the authors and does not necessarily represent the official views of the NIDCR or the NIH. The authors thank Dr. Shorouk Elnagdy for assistance with the mice in the 4-NQO protocol, Drs. Rajaram Gopalakrishnan and Ioannis G. Koutlas, University of Minnesota, for assistance with mouse tissue processing for histopathologic analysis, and Dr. Gay Herzberg for expert editing. Publisher Copyright: © 2022

PY - 2023/2

Y1 - 2023/2

N2 - 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/A9null 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/A9Δ113-114 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/A9null 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.

AB - 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/A9null 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/A9Δ113-114 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/A9null 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.

KW - 53BP1

KW - Calprotectin

KW - Cisplatin

KW - DNA damage

KW - Double-strand breaks

KW - Head and neck cancer

KW - Head and neck squamous cell carcinoma

KW - Radiation

KW - S100A8/A9

KW - γH2AX

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UR - http://www.scopus.com/inward/citedby.url?scp=85146046846&partnerID=8YFLogxK

U2 - 10.1016/j.oraloncology.2022.106304

DO - 10.1016/j.oraloncology.2022.106304

M3 - Article

C2 - 36608459

AN - SCOPUS:85146046846

SN - 1368-8375

VL - 137

JO - Oral Oncology

JF - Oral Oncology

M1 - 106304

ER -

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

Abstract

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Keywords

UN SDGs

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