{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,3]],"date-time":"2026-03-03T05:57:31Z","timestamp":1772517451925,"version":"3.50.1"},"reference-count":28,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2026,2,28]],"date-time":"2026-02-28T00:00:00Z","timestamp":1772236800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100007300","name":"China Medical University","doi-asserted-by":"crossref","award":["CMU113-MF-42"],"award-info":[{"award-number":["CMU113-MF-42"]}],"id":[{"id":"10.13039\/501100007300","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100007300","name":"China Medical University","doi-asserted-by":"crossref","award":["CMU114-ASIA-08"],"award-info":[{"award-number":["CMU114-ASIA-08"]}],"id":[{"id":"10.13039\/501100007300","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100007300","name":"China Medical University","doi-asserted-by":"crossref","award":["CMU114-MF-54"],"award-info":[{"award-number":["CMU114-MF-54"]}],"id":[{"id":"10.13039\/501100007300","id-type":"DOI","asserted-by":"crossref"}]},{"name":"National Science and Technology Council","award":["MOST 111-2314-B-039-027-MY3"],"award-info":[{"award-number":["MOST 111-2314-B-039-027-MY3"]}]},{"name":"National Science and Technology Council","award":["NSTC 114-2314-B-039-023-MY3"],"award-info":[{"award-number":["NSTC 114-2314-B-039-023-MY3"]}]},{"DOI":"10.13039\/501100007632","name":"Changhua Christian Hospital","doi-asserted-by":"crossref","award":["Y-109-0103"],"award-info":[{"award-number":["Y-109-0103"]}],"id":[{"id":"10.13039\/501100007632","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>The speckle-type BTB\/POZ protein (SPOP) is an E3 ubiquitin ligase adaptor typically considered a tumor suppressor, yet its role in head and neck squamous cell carcinoma (HNSCC) remains unclear. This study investigated SPOP expression, arecoline regulation, and its potential as a HNSCC biomarker. SPOP mRNA and its protein were quantified in HNSCC (FaDu, GMN, HSC-3, SAS, and A253) and normal oral epithelial (SG) cell lines via RT-qPCR and Western blot; arecoline\u2019s effect on SG, SAS, and A253 cells was evaluated. SPOP mRNA was analyzed using The Cancer Genome Atlas (TCGA) HNSCC cohort, and protein localization was assessed via immunohistochemistry (IHC) on tissue microarrays. SPOP mRNA was higher in some HNSCC lines; arecoline induced SPOP in SG cells, but not in HNSCC cell lines. TCGA confirmed SPOP mRNA upregulation in tumors correlating with grade. IHC showed SPOP upregulation in HNSCC, particularly in palate and pharynx\/hypopharynx sites. The nuclear SPOP-positive ratio shifted from 12.14 \u00b1 9.82% in normal tissues to 61.26 \u00b1 33.03% in tumors (p &lt; 0.0001), differentiating grades and sites better than total expression. SPOP is upregulated in HNSCC and inducible by arecoline. Enhanced nuclear SPOP localization indicates malignancy and progression, identifying it as a potential HNSCC diagnostic and progression biomarker.<\/jats:p>","DOI":"10.3390\/ijms27052285","type":"journal-article","created":{"date-parts":[[2026,3,2]],"date-time":"2026-03-02T10:24:34Z","timestamp":1772447074000},"page":"2285","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["The Clinical Significance of SPOP Upregulation and Nuclear Accumulation in Head and Neck Squamous Cell Carcinoma"],"prefix":"10.3390","volume":"27","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0893-4185","authenticated-orcid":false,"given":"Yin-Hwa","family":"Shih","sequence":"first","affiliation":[{"name":"Department of Healthcare Administration, Asia University, Taichung 41354, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2504-7708","authenticated-orcid":false,"given":"Nan-Chin","family":"Lin","sequence":"additional","affiliation":[{"name":"Department of Oral and Maxillofacial Surgery, Show Chwan Memorial Hospital, Changhua 500, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yen-Wen","family":"Shen","sequence":"additional","affiliation":[{"name":"School of Dentistry, China Medical University, Taichung 404328, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ming-Gene","family":"Tu","sequence":"additional","affiliation":[{"name":"School of Dentistry, China Medical University, Taichung 404328, Taiwan"},{"name":"Department of Dentistry, China Medical University Hospital, Taichung 40447, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9248-9031","authenticated-orcid":false,"given":"Tong-Hong","family":"Wang","sequence":"additional","affiliation":[{"name":"Tissue Bank, Chang Gung Memorial Hospital at Linkou, Taoyuan 33305, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2558-2209","authenticated-orcid":false,"given":"Yu-Hsin","family":"Tseng","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, Kaohsiung Medical University Hospital, Kaohsiung Medical University, Kaohsiung 80756, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2888-2618","authenticated-orcid":false,"given":"Shih-Min","family":"Hsia","sequence":"additional","affiliation":[{"name":"School of Nutrition and Health Sciences, College of Nutrition, Taipei Medical University, Taipei 110301, Taiwan"},{"name":"Nutrition Research Center, Taipei Medical University Hospital, Taipei 110301, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1759-9120","authenticated-orcid":false,"given":"Tzong-Ming","family":"Shieh","sequence":"additional","affiliation":[{"name":"School of Dentistry, China Medical University, Taichung 404328, Taiwan"},{"name":"Institute of Oral Biology, College of Dentistry, National Yang Ming Chiao Tung University, Taipei 110301, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2026,2,28]]},"reference":[{"key":"ref_1","first-page":"209","article-title":"Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries","volume":"71","author":"Sung","year":"2021","journal-title":"CA Cancer J. Clin."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1520","DOI":"10.1002\/hed.28109","article-title":"Global Incidence, Mortality, and Risk Factors of Stroke in Multi-Modality Head and Neck Cancer Treatment-A Systematic Review and Meta-Analysis","volume":"47","author":"Vasudevan","year":"2025","journal-title":"Head Neck"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"102526","DOI":"10.1016\/j.canep.2024.102526","article-title":"Association of alcohol consumption, betel nut chewing, and cigarette smoking with mortality in patients with head and neck cancer among the Taiwanese population: A nationwide population-based cohort study","volume":"89","author":"Su","year":"2024","journal-title":"Cancer Epidemiol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"489","DOI":"10.1158\/0008-5472.CAN-22-2801","article-title":"Deregulation of SPOP in cancer","volume":"83","author":"Zhang","year":"2023","journal-title":"Cancer Res."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"3946","DOI":"10.1111\/febs.15056","article-title":"The ubiquitin ligase adaptor SPOP in cancer","volume":"286","author":"Cuneo","year":"2019","journal-title":"FEBS J."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"3257","DOI":"10.7150\/jca.25930","article-title":"Functional roles of Speckle-Type Poz (SPOP) Protein in Genomic stability","volume":"9","author":"Wei","year":"2018","journal-title":"J. Cancer"},{"key":"ref_7","first-page":"704","article-title":"SPOP and cancer: A systematic review","volume":"10","author":"Clark","year":"2020","journal-title":"Am. J. Cancer Res."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1063","DOI":"10.1038\/nm.4378","article-title":"Prostate cancer-associated SPOP mutations confer resistance to BET inhibitors through stabilization of BRD4","volume":"23","author":"Dai","year":"2017","journal-title":"Nat. Med."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"657","DOI":"10.1016\/j.celrep.2014.01.013","article-title":"Destruction of full-length androgen receptor by wild-type SPOP, but not prostate-cancer-associated mutants","volume":"6","author":"An","year":"2014","journal-title":"Cell Rep."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"e1687","DOI":"10.1038\/cddis.2015.47","article-title":"Endometrial cancer-associated mutants of SPOP are defective in regulating estrogen receptor-\u03b1 protein turnover","volume":"6","author":"Zhang","year":"2015","journal-title":"Cell Death Dis."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"2","DOI":"10.1186\/s12943-019-1124-x","article-title":"The emerging role of SPOP protein in tumorigenesis and cancer therapy","volume":"19","author":"Song","year":"2020","journal-title":"Mol. Cancer"},{"key":"ref_12","first-page":"5270","article-title":"MiR-373 promotes proliferation and metastasis of oral squamous cell carcinoma by targeting SPOP","volume":"23","author":"Zhang","year":"2019","journal-title":"Eur. Rev. Med. Pharmacol. Sci."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"209","DOI":"10.14712\/23362936.2020.19","article-title":"A Review on Role of Arecoline and Its Metabolites in the Molecular Pathogenesis of Oral Lesions with an Insight into Current Status of Its Metabolomics","volume":"121","author":"Das","year":"2020","journal-title":"Prague Med. Rep."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"2525","DOI":"10.1002\/hed.25718","article-title":"Arecoline-regulated ataxia telangiectasia mutated expression level in oral cancer progression","volume":"41","author":"Tu","year":"2019","journal-title":"Head Neck"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"478","DOI":"10.1091\/mbc.E19-08-0456","article-title":"SPOP is essential for DNA-protein cross-link repair in prostate cancer cells: SPOP-dependent removal of topoisomerase 2A from the topoisomerase 2A-DNA cleavage complex","volume":"31","author":"Watanabe","year":"2020","journal-title":"Mol. Biol. Cell"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1691","DOI":"10.1093\/carcin\/bgu022","article-title":"Speckle-type POZ protein, SPOP, is involved in the DNA damage response","volume":"35","author":"Zhang","year":"2014","journal-title":"Carcinogenesis"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"9891","DOI":"10.1093\/nar\/gky788","article-title":"SPOP promotes transcriptional expression of DNA repair and replication factors to prevent replication stress and genomic instability","volume":"46","author":"Dalgaard","year":"2018","journal-title":"Nucleic Acids Res."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"455","DOI":"10.1016\/j.ccr.2014.02.007","article-title":"SPOP promotes tumorigenesis by acting as a key regulatory hub in kidney cancer","volume":"25","author":"Li","year":"2014","journal-title":"Cancer Cell"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"ar24","DOI":"10.1091\/mbc.E24-04-0198","article-title":"Cul3 substrate adaptor SPOP targets Nup153 for degradation","volume":"36","author":"Ong","year":"2025","journal-title":"Mol. Biol. Cell"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"172","DOI":"10.1038\/s41419-024-06565-1","article-title":"SPOP point mutations regulate substrate preference and affect its function","volume":"15","author":"Deng","year":"2024","journal-title":"Cell Death Dis."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"e189048","DOI":"10.1172\/JCI189048","article-title":"Cancer-associated SPOP mutations enlarge nuclear size and facilitate nuclear envelope rupture upon farnesyltransferase inhibitor treatment","volume":"135","author":"Wang","year":"2025","journal-title":"J. Clin. Investig."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"42845","DOI":"10.1038\/srep42845","article-title":"The CUL3-SPOP-DAXX axis is a novel regulator of VEGFR2 expression in vascular endothelial cells","volume":"7","author":"Sakaue","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1016\/j.molcel.2018.08.027","article-title":"Cancer Mutations of the Tumor Suppressor SPOP Disrupt the Formation of Active, Phase-Separated Compartments","volume":"72","author":"Bouchard","year":"2018","journal-title":"Mol. Cell"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"7635","DOI":"10.1073\/pnas.0408918102","article-title":"Stable X chromosome inactivation involves the PRC1 Polycomb complex and requires histone MACROH2A1 and the CULLIN3\/SPOP ubiquitin E3 ligase","volume":"102","author":"Lund","year":"2005","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"339","DOI":"10.1038\/s41585-020-0314-z","article-title":"The diverse roles of SPOP in prostate cancer and kidney cancer","volume":"17","author":"Wang","year":"2020","journal-title":"Nat. Rev. Urol."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Lin, N.-C., Shih, Y.-H., Chiu, K.-C., Li, P.-J., Yang, H.-W., Lan, W.-C., Hsia, S.-M., Wang, T.-H., and Shieh, T.-M. (2022). Association of rs9679162 Genetic Polymorphism and Aberrant Expression of Polypeptide N-Acetylgalactosaminyltransferase 14 (GALNT14) in Head and Neck Cancer. Cancers, 14.","DOI":"10.3390\/cancers14174217"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"174","DOI":"10.1016\/j.biomed.2013.09.001","article-title":"Leptin induces cell invasion and the upregulation of matrilysin in human colon cancer cells","volume":"3","author":"Lin","year":"2013","journal-title":"BioMedicine"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"113165","DOI":"10.1016\/j.jphotobiol.2025.113165","article-title":"Mechanistic insights into temoporfin-based photodynamic therapy: Ferroptosis as a critical regulator under normoxic and hypoxic conditions in head and neck cancer","volume":"267","author":"Wang","year":"2025","journal-title":"J. Photochem. Photobiol. B Biol."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/5\/2285\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,3,3]],"date-time":"2026-03-03T05:15:11Z","timestamp":1772514911000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/5\/2285"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,2,28]]},"references-count":28,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2026,3]]}},"alternative-id":["ijms27052285"],"URL":"https:\/\/doi.org\/10.3390\/ijms27052285","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,2,28]]}}}