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Article Open Access

Deciphering site‑specific histopathological parameters with potential clinical value in head and neck squamous cell carcinomas

  • Authors:
    • Sofia-Eleni Tzorakoleftheraki
    • Vassiliki Kotoula
    • Georgios Karakatsoulis
    • Konstantinos Markou
    • Stavroula Pervana
    • Konstantinos Vlachtsis
    • Sofia Chrisafi
    • Amanda Psyrri
    • George Fountzilas
    • Prodromos Hytiroglou
    • Triantafyllia Koletsa
  • View Affiliations / Copyright

    Affiliations: Department of Pathology, Aristotle University of Thessaloniki, School of Health Sciences, Faculty of Medicine, 54124 Thessaloniki, Greece, Institute of Applied Biosciences, Centre for Research and Technology Hellas, 57001 Thessaloniki, Greece, Department of Otorhinolaryngology, Papageorgiou Hospital, Aristotle University of Thessaloniki, 56403 Thessaloniki, Greece, Department of Pathology, Papageorgiou Hospital, 56403 Thessaloniki, Greece, First Department of Otorhinolaryngology, Papanikolaou Hospital, 57010 Thessaloniki, Greece, Laboratory of Molecular Oncology, Hellenic Foundation for Cancer Research/Aristotle University of Thessaloniki, 54124 Thessaloniki, Greece, Section of Medical Oncology, Department of Internal Medicine, Attikon University Hospital, Faculty of Medicine, National and Kapodistrian University of Athens School of Medicine, 12462 Athens, Greece
    Copyright: © Tzorakoleftheraki et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 569
    |
    Published online on: October 3, 2025
       https://doi.org/10.3892/ol.2025.15315
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Abstract

In the diagnostic setting of head and neck squamous cell carcinoma (HNSCC), there is an unmet need for robust histological prognosticators, since grading alone is considered to be of disputed clinical relevance. In this context, the present study assessed site‑specific and tumor compartment characteristics as potential histological risk factors in HNSCC. Morphological and immunophenotypic (such as CD8 and PD‑L1) characteristics of tumor cells, the immune microenvironment and the invasive margin (IM) were assessed in 248 patients with HNSCC who were followed up for >20 years, to determine their site specificity and impact on the overall survival of patients. Laryngeal and hypopharyngeal carcinomas were characterized by keratinization, cell cannibalism and anaplastic features; oropharyngeal carcinoma was characterized by a high grade and necrosis; and oral carcinomas was characterized by keratin pearls, eosinophil infiltrates and perineural invasion (P<0.05). CD8 distribution homogeneity, tumor center and/or IM perineural invasion, and sparse lymphocytic host response in the IM (LHR‑IM) were unfavorable prognosticators (P<0.05). Among these parameters, only LHR‑IM and perineural invasion throughout the tumor independently predicted an unfavorable prognosis, along with disease stage. Grade, keratinization, anaplastic features, cell cannibalism, necrosis‑related features, eosinophils, worst pattern of invasion (the most aggressive pattern of tumor cell proliferation found at the invasive front), tumor lymphocytic and CD8+ T‑cell infiltrates, and primary site were not associated with prognosis in the present study. In conclusion, perineural invasion and LHR‑IM were confirmed as histological risk factors in HNSCC, which should be included in pathology reports. To revise the assessment of HNSCC tumor grade, the aforementioned site‑specific histological characteristics may be used in deep learning algorithms.
View Figures

Figure 1

Histopathological features of HNSCC
cases: (A) Cell-in-cell structures (arrows) in a case of laryngeal
carcinoma (H&E ×400). (B) Neutrophils in dyskeratosis
(encircled)/inset: neutrophils in close proximity to dyskeratotic
cell in a case of laryngeal carcinoma (H&E ×400). (C) Necrosis
in keratinization in a case of laryngeal carcinoma (H&E ×200);
Grading heterogeneity with (D) well differentiated and (E)
moderately differentiated areas within a single case of laryngeal
carcinoma (H&E ×200; H&E ×200). (F) Multiple worst patterns
of invasion along the invasive margin within the same case of oral
carcinoma (H&E ×100). (G) Perineural invasion in the invasive
margin of an oral carcinoma (H&E ×100). Heterogeneity in T8
lymphocytic density in an oropharyngeal case presenting (H)
abundant CD8+ T cells in one tissue core of the tumor,
and (I) only a few in the other (CD8 ×200). HNSCC, head and neck
squamous cell carcinoma.

Figure 2

Profile map of histological
parameters focused on keratinization. Grade 1, green; grade 2,
pink; grade 3, dark blue.

Figure 3

Significant associations of overall
survival with CD8 heterogeneity in the entire tumor area as well as
with PNI and LHR in the invasive margin. PNI, perineural invasion;
LHR, lymphocytic host response.
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Tzorakoleftheraki S, Kotoula V, Karakatsoulis G, Markou K, Pervana S, Vlachtsis K, Chrisafi S, Psyrri A, Fountzilas G, Hytiroglou P, Hytiroglou P, et al: Deciphering site‑specific histopathological parameters with potential clinical value in head and neck squamous cell carcinomas. Oncol Lett 30: 569, 2025.
APA
Tzorakoleftheraki, S., Kotoula, V., Karakatsoulis, G., Markou, K., Pervana, S., Vlachtsis, K. ... Koletsa, T. (2025). Deciphering site‑specific histopathological parameters with potential clinical value in head and neck squamous cell carcinomas. Oncology Letters, 30, 569. https://doi.org/10.3892/ol.2025.15315
MLA
Tzorakoleftheraki, S., Kotoula, V., Karakatsoulis, G., Markou, K., Pervana, S., Vlachtsis, K., Chrisafi, S., Psyrri, A., Fountzilas, G., Hytiroglou, P., Koletsa, T."Deciphering site‑specific histopathological parameters with potential clinical value in head and neck squamous cell carcinomas". Oncology Letters 30.6 (2025): 569.
Chicago
Tzorakoleftheraki, S., Kotoula, V., Karakatsoulis, G., Markou, K., Pervana, S., Vlachtsis, K., Chrisafi, S., Psyrri, A., Fountzilas, G., Hytiroglou, P., Koletsa, T."Deciphering site‑specific histopathological parameters with potential clinical value in head and neck squamous cell carcinomas". Oncology Letters 30, no. 6 (2025): 569. https://doi.org/10.3892/ol.2025.15315
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Spandidos Publications style
Tzorakoleftheraki S, Kotoula V, Karakatsoulis G, Markou K, Pervana S, Vlachtsis K, Chrisafi S, Psyrri A, Fountzilas G, Hytiroglou P, Hytiroglou P, et al: Deciphering site‑specific histopathological parameters with potential clinical value in head and neck squamous cell carcinomas. Oncol Lett 30: 569, 2025.
APA
Tzorakoleftheraki, S., Kotoula, V., Karakatsoulis, G., Markou, K., Pervana, S., Vlachtsis, K. ... Koletsa, T. (2025). Deciphering site‑specific histopathological parameters with potential clinical value in head and neck squamous cell carcinomas. Oncology Letters, 30, 569. https://doi.org/10.3892/ol.2025.15315
MLA
Tzorakoleftheraki, S., Kotoula, V., Karakatsoulis, G., Markou, K., Pervana, S., Vlachtsis, K., Chrisafi, S., Psyrri, A., Fountzilas, G., Hytiroglou, P., Koletsa, T."Deciphering site‑specific histopathological parameters with potential clinical value in head and neck squamous cell carcinomas". Oncology Letters 30.6 (2025): 569.
Chicago
Tzorakoleftheraki, S., Kotoula, V., Karakatsoulis, G., Markou, K., Pervana, S., Vlachtsis, K., Chrisafi, S., Psyrri, A., Fountzilas, G., Hytiroglou, P., Koletsa, T."Deciphering site‑specific histopathological parameters with potential clinical value in head and neck squamous cell carcinomas". Oncology Letters 30, no. 6 (2025): 569. https://doi.org/10.3892/ol.2025.15315
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