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NF‑κB‑driven LUZP1 promotes metastasis and chemoresistance in head and neck squamous cell carcinoma

  • Authors:
    • Chen-Yuan Lin
    • Ching-Yun Hsieh
    • Hsin-Chi Lan
    • Ching-Chan Lin
    • Tzu-Ting Chen
    • Wei-Chi Tseng
    • Yung-An Tsou
    • Wei-Chao Chang
  • View Affiliations / Copyright

    Affiliations: School of Pharmacy and Graduate Institute, China Medical University, Taichung 406040, Taiwan, R.O.C., Division of Hematology and Oncology, China Medical University Hospital, Taichung 404327, Taiwan, R.O.C., Center for Molecular Medicine, China Medical University Hospital, Taichung 406040, Taiwan, R.O.C., Department of Otolaryngology‑Head and Neck Surgery, China Medical University Hospital, Taichung 404327, Taiwan, R.O.C.
    Copyright: © Lin et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 110
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    Published online on: April 7, 2026
       https://doi.org/10.3892/or.2026.9115
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Abstract

Head and neck squamous cell carcinoma (HNSCC) remains a highly aggressive malignancy with poor prognosis driven by metastasis and therapeutic resistance. Through comparative proteomic profiling of tumor specimens from patients with long‑ and short‑term survival, the present study identified leucine zipper protein 1 (LUZP1) as one of the most upregulated proteins in tumors from short‑term survivors. Functional assays revealed that LUZP1 knockdown impaired migration, invasion, invadopodia formation and epithelial‑mesenchymal transition, while enhancing sensitivity to docetaxel and cisplatin. Analysis of paired primary and metastatic tumors further confirmed elevated LUZP1 expression in metastatic sites. Mechanistically, NF‑κB inhibition markedly reduced LUZP1 expression, whereas stimulation with IL‑1β or TNF‑α induced its upregulation and rescued the migration defect caused by LUZP1 depletion, implicating NF‑κB as a key upstream regulator. Immunohistochemical analysis of clinical samples demonstrated that high LUZP1 expression was associated with shorter overall and progression‑free survival. Collectively, these findings identify LUZP1 as a novel NF‑κB‑regulated effector that promotes metastasis and chemoresistance and highlight its potential as a prognostic biomarker and therapeutic target in HNSCC.
View Figures

Figure 1

LUZP1 is highly expressed in HNSCC
tumor tissues with short-term survival. (A) KEGG pathway annotation
of differentially upregulatory proteins identified from the
proteomic analysis of tissue specimens of patients with HNSCC with
short-term survival compared with long-term survival. The circle
size indicates the number of mapping proteins in each process, and
the color indicates the P-value of each pathway. (B) Differential
protein expression between short- and long-term survival tissues
using a proteomic volcano plot. The x-axis represents the
log2 fold change in protein abundance and the y-axis
represents the -log10 P-value, indicating statistical
significance. The arrow indicated LUZP1 as one of the most
upregulated proteins in the short-term survival specimen. (C) The
expression of LUZP1 in both long- and short-term survival specimen
was determined by western blot assay. Signal quantification was
measured using ImageJ 1.54 g software (National Institutes of
Health) and the relative intensity was normalized to the long-term
survival specimen group (n=2). β-actin, loading control. (D) The
relative LUZP1 mRNA levels between HNSCC tissues and normal tissues
were analyzed on the UALCAN website using The Cancer Genome Atlas
database. (E) Colony formation assay in OECM-1 cells with or
without LUZP1 knockdown. Signal quantification using crystal violet
extract was measured by colorimetric analysis at 570 nm. The
relative signal intensities were normalized to the shControl (n=3).
(E) Data are displayed as the means ± SD. For statistical analysis,
(A-C) one-way ANOVA followed by Dunnett's post hoc. **P<0.01.
LUZP1, leucine zipper protein 1; HNSCC, head and neck squamous cell
carcinoma; KEGG, Kyoto Encyclopedia of Genes and Genomes; sh, short
hairpin; Ctl., control.

Figure 2

LUZP1 enhances the metastatic
abilities of HNSCC cells. (A) cell migration and (B) invasion assay
in OECM-1 cells with or without LUZP1 knockdown. Signal
quantification using crystal violet extract was measured by
colorimetric analysis at 570 nm. The relative signal intensities
were normalized to the shControl (n=3). (C) The representative
images for 3D invasion assay for tumor spheroids in SAS cells with
or without LUZP1 knockdown. A radial outgrowth distance approach is
used for quantification of 3D tumor spheroid invasion. Scale bar,
500 µm. (D) Invadopodia-like protrusive structures were visualized
by F-actin staining using phalloidin (cat. no. R415; Invitrogen;
Thermo Fisher Scientific, Inc.), which are indicated by the arrow.
Scale bar, 20 µm. (E) The expression of LUZP1, E-cadherin and
vimentin in OECM-1 cells with or without LUZP1 knockdown was
determined by western blot assay. β-actin, loading control. (F) The
expression of LUZP1 in both primary and metastatic tumors of a
patient with HNSCC was examined using immunohistochemical analysis.
Scale bar, 50 µm. Data are shown as the means ± SD. For statistical
analyses, (A and B) a 2-tailed unpaired Student's t-test; (C)
one-way ANOVA with Tukey's post hoc test. **P<0.01. LUZP1,
leucine zipper protein 1; HNSCC, head and neck squamous cell
carcinoma; sh, short hairpin; Ctl., control.

Figure 3

LUZP1 increases chemoresistance of
head and neck squamous cell carcinoma cells. (A) Immunofluorescent
analysis was conducted to examine LUZP1 expression in
DTX-resistant, CIS-resistant and the control OECM-1 cells. Scale
bar, 20 µm. Cell viability of OECM-1 and SAS with or without LUZP1
knockdown treated with the indicated doses of (B) DTX and (C) CIS
was analyzed by MTT assay (n=3). For statistical analyses, (B and
C) one-way ANOVA with Tukey's post hoc test. *P<0.05;
**P<0.01. LUZP1, leucine zipper protein 1; sh, short hairpin;
DTX, docetaxel; CIS, cisplatin.

Figure 4

NF-κB signaling activation promotes
LUZP1 expression in HNSCC cells. (A) The expression of LUZP1 in
FaDu, OECM-1 and SAS cells treated with SB431542 (10 µM), LY294002
(10 µM), Rapamycin (10 µM), BAY 11–7085 (5 µM) or YC-1 (30 µM) was
determined by western blot assay. Signal quantification was
measured using ImageJ 1.54 g software (National Institutes of
Health) and the relative intensity was normalized to untreated
control. The red dashed line represents the normalized value as 1.
(B) The expression of LUZP1 in FaDu, OECM-1 and SAS cells with or
without BAY 11–7085 was validated by western blot assay. (C)
Spearman's monotonic correlation between LUZP1 and NFKB1 or NFKB2
expression in HNSCC was analyzed using The Cancer Genome Atlas
RNA-Sequencing database on the GEPIA server. (D) Protein expression
of NF-κB p65 and LUZP1 in OECM-1 and SAS cells with NF-κB p65
knockdown (+) or control shRNA -), as determined by western blot
analysis. β-actin, loading control. (E) IC50 values of
docetaxel in OECM-1 and SAS cells with or without NF-κB p65
knockdown. (F) IC50 values of cisplatin in OECM-1 and
SAS cells with or without NF-κB p65 knockdown. The expression of
LUZP1 in different HNSCC cells treated with (G) IL-1β (3 ng/ml) or
(H) TNFα (10 ng/ml) for the indicated times was determined by
western blot assay. β-actin, loading control. (I) Transwell cell
migration assay was conducted using OECM-1 cells with or without
LUZP1 knockdown in the presence or absence of TNFα (10 ng/ml)
treatment. Signal quantification using crystal violet extract was
measured by colorimetric analysis at 570 nm, and the relative
signal intensities were normalized to untreated shControl (shLUZP1,
-; TNFα, -) (n=3). (J) Genomic visualization of the human LUZP1
locus (GRCh38/hg38) showing RefSeq-curated exon annotations, NF-κB
RelA ChIP-seq binding signals in FaDu cells (ReMap), layered
H3K27ac ChIP-seq profiles from ENCODE cell lines, and GeneHancer
regulatory element annotations. Red boxes denote promoter regions
and gray boxes indicate putative enhancers. Blue vertical bars mark
the locations of ChIP-qPCR primer sets designed for experimental
validation. (K) ChIP-qPCR analysis showing increased NF-κB (RelA)
occupancy at the LUZP1 promoter in response to TNF-α treatment. For
statistical analyses, (E and F) a 2-tailed unpaired Student's
t-test; (I) a factorial two-way ANOVA, followed by Tukey's
Honestly Significant Difference post hoc test. **P<0.01. TPM,
transcripts per million; ChIP-seq, chromatin immunoprecipitation
sequencing; LUZP1, leucine zipper protein 1; sh, short hairpin.

Figure 5

LUZP1 is associated with poor
survival of patients with head and neck squamous cell carcinoma.
(A) Representative immunohistochemical images demonstrating weak
LUZP1 signal (LUZP 1−; a score=0 or 1) and strong LUZP1
signal (LUZP 1+; score=2 or 3) in pathologic specimens
of patients with HNSCC. Correlations between LUZP1 expression and
(B) OS or (C) PFS were analyzed according to LUZP1 signals using
the Kaplan-Meier method. For statistical analyses, (B and C)
log-rank test. OS, overall survival; PFS, progression free
survival; LUZP1, leucine zipper protein 1; ChIP-seq, chromatin
immunoprecipitation sequencing.
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Copy and paste a formatted citation
Spandidos Publications style
Lin C, Hsieh C, Lan H, Lin C, Chen T, Tseng W, Tsou Y and Chang W: NF‑&kappa;B‑driven LUZP1 promotes metastasis and chemoresistance in head and neck squamous cell carcinoma. Oncol Rep 55: 110, 2026.
APA
Lin, C., Hsieh, C., Lan, H., Lin, C., Chen, T., Tseng, W. ... Chang, W. (2026). NF‑&kappa;B‑driven LUZP1 promotes metastasis and chemoresistance in head and neck squamous cell carcinoma. Oncology Reports, 55, 110. https://doi.org/10.3892/or.2026.9115
MLA
Lin, C., Hsieh, C., Lan, H., Lin, C., Chen, T., Tseng, W., Tsou, Y., Chang, W."NF‑&kappa;B‑driven LUZP1 promotes metastasis and chemoresistance in head and neck squamous cell carcinoma". Oncology Reports 55.6 (2026): 110.
Chicago
Lin, C., Hsieh, C., Lan, H., Lin, C., Chen, T., Tseng, W., Tsou, Y., Chang, W."NF‑&kappa;B‑driven LUZP1 promotes metastasis and chemoresistance in head and neck squamous cell carcinoma". Oncology Reports 55, no. 6 (2026): 110. https://doi.org/10.3892/or.2026.9115
Copy and paste a formatted citation
x
Spandidos Publications style
Lin C, Hsieh C, Lan H, Lin C, Chen T, Tseng W, Tsou Y and Chang W: NF‑&kappa;B‑driven LUZP1 promotes metastasis and chemoresistance in head and neck squamous cell carcinoma. Oncol Rep 55: 110, 2026.
APA
Lin, C., Hsieh, C., Lan, H., Lin, C., Chen, T., Tseng, W. ... Chang, W. (2026). NF‑&kappa;B‑driven LUZP1 promotes metastasis and chemoresistance in head and neck squamous cell carcinoma. Oncology Reports, 55, 110. https://doi.org/10.3892/or.2026.9115
MLA
Lin, C., Hsieh, C., Lan, H., Lin, C., Chen, T., Tseng, W., Tsou, Y., Chang, W."NF‑&kappa;B‑driven LUZP1 promotes metastasis and chemoresistance in head and neck squamous cell carcinoma". Oncology Reports 55.6 (2026): 110.
Chicago
Lin, C., Hsieh, C., Lan, H., Lin, C., Chen, T., Tseng, W., Tsou, Y., Chang, W."NF‑&kappa;B‑driven LUZP1 promotes metastasis and chemoresistance in head and neck squamous cell carcinoma". Oncology Reports 55, no. 6 (2026): 110. https://doi.org/10.3892/or.2026.9115
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