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Article Open Access

Inhibitory role of angiopoietin‑like 4 for cancer progression in oropharyngeal squamous cell carcinoma

  • Authors:
    • Takuya Mikoshiba
    • Mariko Sekimizu
    • Nana Nakahara
    • Shin Saito
    • Keisuke Yoshihama
    • Ryoto Nagai
    • Takeyuki Kono
    • Hiroyuki Ozawa
  • View Affiliations / Copyright

    Affiliations: Department of Otolaryngology, Head and Neck Surgery, Keio University School of Medicine, Tokyo 160‑8582, Japan
    Copyright: © Mikoshiba et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY 4.0].
  • Article Number: 117
    |
    Published online on: April 20, 2026
       https://doi.org/10.3892/or.2026.9122
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Abstract

The mortality rate of head and neck squamous cell carcinoma (HNSCC), one of the most prevalent types of cancer, has remained unchanged despite advances in treatment strategies. Angiopoietin‑like 4 (ANGPTL4) exhibits both pro‑ and anti‑tumorigenic roles in various cancers depending on the tissue context. The present study investigated the role of ANGPTL4 expression in oropharyngeal squamous cell carcinoma (OPSCC). Data from 137 patients with OPSCC who underwent initial treatment were retrospectively analyzed. ANGPTL4 expression in tumor tissues was determined via immunohistochemistry. Survival outcomes [overall survival (OS) and disease‑free survival (DFS)] and clinicopathological correlations were evaluated. Tumor cell proliferation was assessed using an CellTiter‑Glo 2.0 luminescence‑based cell viability assay, immunofluorescence staining, and reverse transcription‑quantitative polymerase chain reaction following small interfering RNA‑mediated knockdown in a HNSCC cell line. Gene set enrichment analysis (GSEA) was performed using data from The Cancer Genome Atlas. ANGPTL4 expression (≥7.7%) in patients with OPSCC was significantly associated with improved OS and DFS. Multivariate analysis confirmed that low ANGPTL4 expression was an independent prognostic factor for OS [hazard ratio (HR)=3.676, 95% confidence interval (CI)=1.678‑8.056; P=0.001) and DFS (HR=2.959, 95% CI=1.533‑5.713; P=0.001)]. FaDu cells with ANGPTL4 knockdown demonstrated significantly increased proliferation compared with negative controls in the CellTiter‑Glo 2.0 assay (P=0.010), accompanied by a significant increase in Ki‑67 expression as revealed by immunofluorescence staining (P=0.021). The relative ANGPTL4 mRNA expression levels decreased to 38%, whereas those of MKI67 increased significantly. GSEA revealed significant enrichment of cell cycle progression signatures in cases with low ANGPTL4 expression. ANGPTL4 expression was significantly associated with a favorable prognosis in OPSCC, and its knockdown increased proliferative activity in FaDu cells. Thus, ANGPTL4 may serve as a prognostic biomarker in OPSCC. Further in vivo studies are warranted to clarify causality and assess the therapeutic potential of ANGPTL4 targeting in OPSCC.
View Figures

Figure 1

Immunohistochemical staining of
oropharyngeal squamous cell carcinoma. Representative cases with
(A) positive and (B) negative staining of angiopoietin-like 4 are
shown (lower magnification, ×100; scale bar, 250 µm; higher
magnification, ×400; scale bar, 50 µm).

Figure 2

(A) Overall survival and (B)
disease-free survival of patients with oropharyngeal squamous cell
carcinoma based on ANGPTL4. ANGPTL4, angiopoietin-like 4.

Figure 3

Fluorescence intensities of
anti-angiopoietin-like 4 (red), anti-Ki-67 (green), and Hoechst
(blue) for the FaDu cells. (A-C) The fluorescence intensities of
the FaDu cells transfected with angiopoietin-like 4 were
significantly different from those of the control. *P<0.05.

Figure 4

CellTiter-Glo 2.0 luminescence assay
revealed a significant increase in cell viability following
angiopoietin-like 4 knockdown relative to the controls (set as 1.0)
(P=0.010). *P<0.05.

Figure 5

Relative gene expression levels of
various mRNAs in the FaDu cells transfected with
angiopoietin-like 4 small interfering RNA were determined by
reverse transcription-quantitative PCR. The angiopoietin-like
4 level decreased by 38%, and MKI67 expression increased
significantly. The expression of BAX decreased, and that of
BCL2 increased. CASP3 expression also increased in
the angiopoietin-like 4 knockdown cells.

Figure 6

Correlations between ANGPTL4
expression and overall survival based on The Cancer Genome Atlas
data for (A) oropharyngeal, (B) tongue and (C) laryngeal squamous
cell carcinoma. The median FPKM-UQ value for ANGPTL4 was used as
the cut-off to divide the groups into high and low. (D) The mean
ANGPTL4 expression in head and neck squamous cell carcinoma was
significantly different (P=0.042). ANGPTL4, angiopoietin-like
4.
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Copy and paste a formatted citation
Spandidos Publications style
Mikoshiba T, Sekimizu M, Nakahara N, Saito S, Yoshihama K, Nagai R, Kono T and Ozawa H: Inhibitory role of angiopoietin‑like 4 for cancer progression in oropharyngeal squamous cell carcinoma. Oncol Rep 55: 117, 2026.
APA
Mikoshiba, T., Sekimizu, M., Nakahara, N., Saito, S., Yoshihama, K., Nagai, R. ... Ozawa, H. (2026). Inhibitory role of angiopoietin‑like 4 for cancer progression in oropharyngeal squamous cell carcinoma. Oncology Reports, 55, 117. https://doi.org/10.3892/or.2026.9122
MLA
Mikoshiba, T., Sekimizu, M., Nakahara, N., Saito, S., Yoshihama, K., Nagai, R., Kono, T., Ozawa, H."Inhibitory role of angiopoietin‑like 4 for cancer progression in oropharyngeal squamous cell carcinoma". Oncology Reports 55.6 (2026): 117.
Chicago
Mikoshiba, T., Sekimizu, M., Nakahara, N., Saito, S., Yoshihama, K., Nagai, R., Kono, T., Ozawa, H."Inhibitory role of angiopoietin‑like 4 for cancer progression in oropharyngeal squamous cell carcinoma". Oncology Reports 55, no. 6 (2026): 117. https://doi.org/10.3892/or.2026.9122
Copy and paste a formatted citation
x
Spandidos Publications style
Mikoshiba T, Sekimizu M, Nakahara N, Saito S, Yoshihama K, Nagai R, Kono T and Ozawa H: Inhibitory role of angiopoietin‑like 4 for cancer progression in oropharyngeal squamous cell carcinoma. Oncol Rep 55: 117, 2026.
APA
Mikoshiba, T., Sekimizu, M., Nakahara, N., Saito, S., Yoshihama, K., Nagai, R. ... Ozawa, H. (2026). Inhibitory role of angiopoietin‑like 4 for cancer progression in oropharyngeal squamous cell carcinoma. Oncology Reports, 55, 117. https://doi.org/10.3892/or.2026.9122
MLA
Mikoshiba, T., Sekimizu, M., Nakahara, N., Saito, S., Yoshihama, K., Nagai, R., Kono, T., Ozawa, H."Inhibitory role of angiopoietin‑like 4 for cancer progression in oropharyngeal squamous cell carcinoma". Oncology Reports 55.6 (2026): 117.
Chicago
Mikoshiba, T., Sekimizu, M., Nakahara, N., Saito, S., Yoshihama, K., Nagai, R., Kono, T., Ozawa, H."Inhibitory role of angiopoietin‑like 4 for cancer progression in oropharyngeal squamous cell carcinoma". Oncology Reports 55, no. 6 (2026): 117. https://doi.org/10.3892/or.2026.9122
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