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High expression of CDKN2A in oral squamous cell carcinoma is associated with distant metastasis: Cell‑free DNA as a new prognostic feature

  • Authors:
    • Natsuki Eboshida
    • Atsuko Hamada
    • Asami Tanigawa
    • Mirai Higaki
    • Fumitaka Obayashi
    • Nanako Ito
    • Sachiko Yamasaki
    • Ryouji Tani
    • Tomoaki Shintani
    • Koichi Koizumi
    • Souichi Yanamoto
  • View Affiliations / Copyright

    Affiliations: Department of Oral Oncology, Graduate School of Biomedical and Health Sciences, Hiroshima University, Hiroshima 734‑8553, Japan, Centre of Oral Clinical Examination, Hiroshima University Hospital, Hiroshima 734‑8553, Japan
    Copyright: © Eboshida et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 111
    |
    Published online on: October 17, 2025
       https://doi.org/10.3892/mco.2025.2906
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Abstract

Oral squamous cell carcinoma (OSCC) shows a high incidence and is associated with a higher rate of mortality, particularly in developing countries. Although the development of treatment strategies has been remarkable, the 5‑year survival rate remains low (often <50%). Surgery and chemoradiotherapy are the standard treatments; however, there are limited factors that can predict metastasis. Liquid biopsy is promising in treating various cancers, including OSCC. In previous studies on OSCC, associations were identified among circulating tumour cell clusters, cell‑free DNA (cfDNA) size/quantity and prognosis. The present study aimed to explore the role of CDKN2A and TP53 in OSCC using immunohistochemistry. Next‑generation sequencing analysis of both cfDNA and tissues from two patients with poor prognoses was performed, and two common mutated genes, TP53 and CDKN2A, were detected. The CDKN2A‑positive group showed metastasis in 50% of cases and shorter overall survival in OSCC. Overall, CDKN2A protein expression is a potential prognostic marker for OSCC.
View Figures

Figure 1

Flow chart of OSCC case selection with
radical surgery. Of the 53 patients who visited the Department of
Oral and Maxillofacial Surgery at Hiroshima University Hospital
between January 2014 and December 2023, 481 underwent radical
surgery. Among the 481 patients, 65 were pathologically confirmed
as pN(+). Among the pN(+) patients, CDKN2A was positive in 18.5%
(12/65) and negative in 81.5% (53/65). Among the CDKN2A-positive
group, 50% (6/12) showed metastasis, whereas 17% (9/53) of patients
in the CDKN2A-negative group showed metastasis. OSCC, oral squamous
cell carcinoma; pN, pathological lymph node metastasis; cN,
clinical lymph node metastasis.

Figure 2

Detection of mutations in cfDNA and
tumour. A total of 54 genetic mutations were identified in the
cfDNA of patient 1, and 52 mutations were identified in patient 2.
Some of these mutations overlapped with those previously detected
in the tumour tissues of each patient. Notably, two genes,
CDKN2A and TP53, were common in patient 1 and patient
2. cfDNA, cell-free DNA; Pt. 1, patient 1; Pt. 2, patient 2.

Figure 3

Images of IHC. IHC staining images of
CDKN2 and TP53 in tumor and adjacent normal tissues. Tumor tissues
are outlined, and the boundary between tumor and normal tissue is
clearly marked with a demarcation line. Scale bars, 50 µm. (A)
CDKN2A-positive (≥70%), (B) CDKN2A-negative (<70%), (C)
TP53-positive (≥60%) and (D) TP53-negative (<60%). IHC,
immunohistochemistry.

Figure 4

Relationship between IHC result for
CDKN2A/TP53 and the 5-year survival rate. Kaplan-Meier survival
curves for OS based on IHC results for CDKN2A/TP53 in OSCC tissues.
Each graph shows the OS of patients with OSCC, with positive cases
(blue line) and negative cases (red line) in the (A) CDKN2A and (B)
TP53 groups. Statistical significance was assessed using the
log-rank test. Statistical significance was set at P<0.05
(statically significant). (A) Median OS was 16.6 months (95% CI,
11.1-34.9) in the CDKN2A-positive group and 21.0 months (95% CI,
16.0-63.5) in the P16-negative group (P=0.002 log-rank test). (B)
Median overall survival was 34.9 months in the TP53-negative group
and 21.3 months in the TP53-positive group. The survival difference
was not statistically significant (P=0.97 log-rank test). OS,
overall survival; IHC, immunohistochemistry; OSCC, oral squamous
cell carcinoma; CI, confidence intervals.

Figure 5

A new risk factor for OSCC. This
figure was drawn by editing the Clinical Practice Guidelines for
Oral Cancer 2023(24). CDKN2A may
be a new risk factor for OSCC treatment or management. OSCC, oral
squamous cell carcinoma; RT, radiotherapy.
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Copy and paste a formatted citation
Spandidos Publications style
Eboshida N, Hamada A, Tanigawa A, Higaki M, Obayashi F, Ito N, Yamasaki S, Tani R, Shintani T, Koizumi K, Koizumi K, et al: High expression of CDKN2A in oral squamous cell carcinoma is associated with distant metastasis: Cell‑free DNA as a new prognostic feature. Mol Clin Oncol 23: 111, 2025.
APA
Eboshida, N., Hamada, A., Tanigawa, A., Higaki, M., Obayashi, F., Ito, N. ... Yanamoto, S. (2025). High expression of CDKN2A in oral squamous cell carcinoma is associated with distant metastasis: Cell‑free DNA as a new prognostic feature. Molecular and Clinical Oncology, 23, 111. https://doi.org/10.3892/mco.2025.2906
MLA
Eboshida, N., Hamada, A., Tanigawa, A., Higaki, M., Obayashi, F., Ito, N., Yamasaki, S., Tani, R., Shintani, T., Koizumi, K., Yanamoto, S."High expression of CDKN2A in oral squamous cell carcinoma is associated with distant metastasis: Cell‑free DNA as a new prognostic feature". Molecular and Clinical Oncology 23.6 (2025): 111.
Chicago
Eboshida, N., Hamada, A., Tanigawa, A., Higaki, M., Obayashi, F., Ito, N., Yamasaki, S., Tani, R., Shintani, T., Koizumi, K., Yanamoto, S."High expression of CDKN2A in oral squamous cell carcinoma is associated with distant metastasis: Cell‑free DNA as a new prognostic feature". Molecular and Clinical Oncology 23, no. 6 (2025): 111. https://doi.org/10.3892/mco.2025.2906
Copy and paste a formatted citation
x
Spandidos Publications style
Eboshida N, Hamada A, Tanigawa A, Higaki M, Obayashi F, Ito N, Yamasaki S, Tani R, Shintani T, Koizumi K, Koizumi K, et al: High expression of CDKN2A in oral squamous cell carcinoma is associated with distant metastasis: Cell‑free DNA as a new prognostic feature. Mol Clin Oncol 23: 111, 2025.
APA
Eboshida, N., Hamada, A., Tanigawa, A., Higaki, M., Obayashi, F., Ito, N. ... Yanamoto, S. (2025). High expression of CDKN2A in oral squamous cell carcinoma is associated with distant metastasis: Cell‑free DNA as a new prognostic feature. Molecular and Clinical Oncology, 23, 111. https://doi.org/10.3892/mco.2025.2906
MLA
Eboshida, N., Hamada, A., Tanigawa, A., Higaki, M., Obayashi, F., Ito, N., Yamasaki, S., Tani, R., Shintani, T., Koizumi, K., Yanamoto, S."High expression of CDKN2A in oral squamous cell carcinoma is associated with distant metastasis: Cell‑free DNA as a new prognostic feature". Molecular and Clinical Oncology 23.6 (2025): 111.
Chicago
Eboshida, N., Hamada, A., Tanigawa, A., Higaki, M., Obayashi, F., Ito, N., Yamasaki, S., Tani, R., Shintani, T., Koizumi, K., Yanamoto, S."High expression of CDKN2A in oral squamous cell carcinoma is associated with distant metastasis: Cell‑free DNA as a new prognostic feature". Molecular and Clinical Oncology 23, no. 6 (2025): 111. https://doi.org/10.3892/mco.2025.2906
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