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Suppressive role of SCN4B in the epithelial‑mesenchymal transition of lung adenocarcinoma

  • Authors:
    • Mengnan Guo
    • He Li
    • Zhifang Zhao
    • Yuanyuan Wang
    • Jiaqi Tang
    • Binghan Zhong
    • Yunxia Zhao
  • View Affiliations / Copyright

    Affiliations: Department of Respiratory Medicine, Hebei Medical University Third Hospital, Shijiazhuang, Hebei 050051, P.R. China, Department of Respiratory Medicine, Xiangyang Central Hospital, Affiliated Hospital of Hubei University of Arts and Science, Xiangyang, Hubei 441021, P.R. China
    Copyright: © Guo et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 68
    |
    Published online on: February 10, 2026
       https://doi.org/10.3892/or.2026.9073
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Abstract

The poor prognosis and high mortality rate of non‑small cell lung cancer are largely driven by its aggressive migratory and invasive behavior. Epithelial‑mesenchymal transition (EMT) is a central mechanism conferring these malignant traits. The present study examined the expression profile of the sodium channel β4 subunit (SCN4B) in lung adenocarcinoma (LUAD) and explored its regulatory role in EMT. Transcriptomic data from The Cancer Genome Atlas were analyzed to compare SCN4B expression between LUAD and normal tissues, and to assess its relationship with TNM clinical stage (I‑IV), overall survival and diagnostic performance using non‑parametric tests, Kaplan‑Meier analysis and receiver operating characteristic curves, respectively. Functional enrichment analysis, including Gene Ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis, and immune cell infiltration profiling were performed on SCN4B‑associated differentially expressed genes. In vitro, the A549 and H1299 LUAD cell lines were engineered to overexpress SCN4B. Viability, migration, invasion and apoptosis were evaluated using Cell Counting Kit‑8 assays, wound healing assays, Transwell assays and flow cytometry. In addition, western blotting was conducted to assess EMT markers, including E‑cadherin, N‑cadherin, Vimentin and Snail. The results demonstrated that SCN4B expression was markedly reduced in LUAD tissues and low SCN4B expression was associated with unfavorable clinical outcomes. KEGG analysis revealed enrichment of SCN4B‑related genes in the ‘cell adhesion molecules’ pathway, and SCN4B expression levels differed markedly between TNM tumor (T) pathologic stages T1 and T2. Furthermore, SCN4B overexpression suppressed viability, migration and invasion of A549 and H1299 cells, while promoting apoptosis. Western blotting demonstrated upregulation of E‑cadherin, and downregulation of N‑cadherin, Vimentin and Snail in the SCN4B overexpression group compared with the empty vector group, indicating inhibition of EMT. In conclusion, low SCN4B expression was associated with poor prognosis in LUAD. Notably, restoring SCN4B levels suppressed LUAD cell viability, migration and invasion in vitro, accompanied by inhibition of EMT. These findings highlighted SCN4B as a potential tumor suppressor and a promising therapeutic target for LUAD.
View Figures

Figure 1

Proposed mechanism of SCN4B in
the regulation of EMT in lung adenocarcinoma. All activating
signaling pathways are indicated by blue solid arrows, while all
inhibitory pathways are represented by red T-shaped termination
lines. SCN4B, sodium channel β4 subunit; EMT,
epithelial-mesenchymal transition; CDH2, N-cadherin; HDAC, histone
deacetylase.

Figure 2

Differential expression analysis of
SCN4B in LUAD. (A) Differential expression analysis of
SCN4B in LUAD and adjacent non-tumorous tissue. A paired
Student's t-test was used. (B) Gene Expression Omnibus cohort
(GSE31210; Affymetrix GPL570): Comparison of SCN4B
microarray intensity between tumor and normal lung tissues. Welch's
t-test was used. (C) Diagnostic receiver operating characteristic
curve analysis of SCN4B. ***P<0.001,
****P<0.0001. AUC, area under the curve; SCN4B, sodium
channel β4 subunit; LUAD, lung adenocarcinoma; TPM, transcripts per
million; TPR, true positive rate; FPR, false positive rate.

Figure 3

Differentially expressed genes
associated with SCN4B in lung adenocarcinoma. (A) Volcano
plot illustrating SCN4B-associated differentially expressed genes
in the dataset. Red dots represent upregulated genes, blue dots
indicate downregulated genes and gray dots denote non-significant
genes. (B) Spearman's rank correlation analysis between
SCN4B and selected genes. P.adj, adjusted P-value;
SCN4B, sodium channel β4 subunit; TPM, transcripts per
million.

Figure 4

Functional enrichment analysis of
SCN4B-associated differentially expressed genes. (A) Gene Ontology
functional enrichment analysis based on differences. (B) KEGG
functional enrichment analysis. KEGG, Kyoto Encyclopedia of Genes
and Genomes; BP, biological process; CC, cellular component; MF,
molecular function; P adj, adjusted P-value.

Figure 5

Analysis of immune infiltration
related to SCN4B. (A) Bubble plot of immune infiltration
analysis. (B) Correlation analysis of SCN4B expression with
tumor purity, M2 macrophage infiltration (quanTIseq) and NK cell
infiltration (quanTIseq). Spearman correlation analysis was
performed; Ρ-values and P-values are shown. SCN4B, sodium
channel β4 subunit; NK, natural killer; quanTIseq, quantification
of the tumor immune contexture from RNA sequencing; TPM,
transcripts per million.

Figure 6

SCN4B expression in lung
cancer tissues and normal tissues. Immunohistochemical staining
images of SCN4B expression in normal lung tissue and lung
adenocarcinoma tumor tissue were obtained from the online Human
Protein Atlas database (https://www.proteinatlas.org/). The magnification of
the first row of images is ×100; scale bar, 200 µm; the
magnification of the second row of images is ×400; scale bar, 50
µm. SCN4B, sodium channel β4 subunit.

Figure 7

Prognostic analysis and clinical
association analysis of SCN4B. (A) Kaplan-Meier survival
curve for overall survival stratified by SCN4B expression.
(B) Kaplan-Meier survival curve for disease-specific survival based
on SCN4B expression levels. (C) Kaplan-Meier survival curve
for the progression-free interval in patients with high vs. low
SCN4B expression. (D) Comparison of SCN4B expression
across T stages (T1-T4). (E) SCN4B expression in patients
with or without lymph node metastasis. (F) SCN4B expression
in patients with or without distant metastasis. (G) SCN4B
expression across different tumor stages. (D and E) Kruskal-Wallis
test. (F and G) Wilcoxon rank-sum test. ***P<0.001. HR,
hazard ratio; ns, not significant (P>0.05); SCN4B,
sodium channel β4 subunit; TPM, transcripts per million.

Figure 8

Prognostic association of
SCN4B in the GSE31210 cohort. Kaplan-Meier curves were
generated by stratifying patients into high and low SCN4B
expression groups using the median expression value as the cutoff.
The ‘number at risk’ table below the plot indicates the counts
remaining at risk at each time point. Groups were compared using
the two-sided log-rank test (P=0.0042). SCN4B, sodium
channel β4 subunit.

Figure 9

Basal expression of SCN4B across lung
cell lines. (A) Representative western blot images: Lane 1, normal
lung cells; lanes 2–6, lung cancer cell lines. GAPDH served as the
loading control. For densitometric quantification, SCN4B was
normalized to GAPDH and further normalized to the normal cells
(=1). (B) Reverse transcription-quantitative PCR validation in the
same panel of distinct cell lines as in (A) (normalized to GAPDH;
2−ΔΔCq). One-way ANOVA with Dunnett's post hoc test was
used for statistical analysis. N=3 independent experiments.
*P<0.05, **P<0.01, ****P<0.0001. ns,
not significant (P>0.05); SCN4B, sodium channel β4
subunit.

Figure 10

Construction of SCN4B
overexpression models in A549 and H1299 lung adenocarcinoma cell
lines. After transfection with overexpression plasmids for 48 h in
A549 and H1299 cells, the relative mRNA expression level changes of
SCN4B in the cells were detected by reverse
transcription-quantitative PCR. Data were analyzed using a
two-tailed unpaired Student's t-test. N=3 independent experiments.
**P<0.01. SCN4B, sodium channel β4 subunit; oe,
overexpression.

Figure 11

SCN4B induces apoptosis and
inhibits cell viability. (A) Changes in the viability of A549 and
H1299 cells at different time points after transfection (0, 24, 48
and 72 h) were detected using a Cell Counting Kit-8 assay. Data
were analyzed using two-way ANOVA with Bonferroni's post hoc test.
Changes in the proportion of apoptotic cells after 48 h of
transfection in (B) A549 and (C) H1299 cells labeled with
PI/annexin V-FITC double staining were detected by flow cytometry.
Data were analyzes using a two-tailed unpaired Student's t-test.
N=3 independent experiments. *P<0.05, **P<0.01,
***P<0.001, ****P<0.0001. ns, not significant
(P>0.05); SCN4B, sodium channel β4 subunit; oe,
overexpression; OD, optical density.

Figure 12

SCN4B can inhibit the invasion
and migration of lung cancer cells. (A) Changes in the invasion of
A549 and H1299 LUAD cells 48 h after transfection with SCN4B
overexpression plasmid were detected using a Transwell assay
(magnification, ×100; scale bar, 100 µm). (B) Changes in the
migration of A549 and H1299 LUAD cells 48 h after transfection with
SCN4B overexpression plasmid were detected using a cell
scratch assay (magnification, ×100; scale bar, 200 µm). Data were
analyzed using a two-tailed unpaired Student's t-test. N=3
independent experiments. *P<0.05, **P<0.01,
***P<0.001. SCN4B, sodium channel β4 subunit; oe,
overexpression; LUAD, lung adenocarcinoma.

Figure 13

SCN4B can inhibit the
epithelial-mesenchymal transition process of LUAD cells. The
relative protein expression levels of SCN4B, E-cadherin,
N-cadherin, Vimentin and Snail in A549 and H1299 LUAD cells 48 h
after transfection with SCN4B overexpression plasmid were detected
by western blotting. Data were analyzed using a two-tailed unpaired
Student's t-test for each protein. N=3 independent experiments.
****P<0.0001. SCN4B, sodium channel β4 subunit; oe,
overexpression; LUAD, lung adenocarcinoma.
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Copy and paste a formatted citation
Spandidos Publications style
Guo M, Li H, Zhao Z, Wang Y, Tang J, Zhong B and Zhao Y: Suppressive role of SCN4B in the epithelial‑mesenchymal transition of lung adenocarcinoma. Oncol Rep 55: 68, 2026.
APA
Guo, M., Li, H., Zhao, Z., Wang, Y., Tang, J., Zhong, B., & Zhao, Y. (2026). Suppressive role of SCN4B in the epithelial‑mesenchymal transition of lung adenocarcinoma. Oncology Reports, 55, 68. https://doi.org/10.3892/or.2026.9073
MLA
Guo, M., Li, H., Zhao, Z., Wang, Y., Tang, J., Zhong, B., Zhao, Y."Suppressive role of SCN4B in the epithelial‑mesenchymal transition of lung adenocarcinoma". Oncology Reports 55.4 (2026): 68.
Chicago
Guo, M., Li, H., Zhao, Z., Wang, Y., Tang, J., Zhong, B., Zhao, Y."Suppressive role of SCN4B in the epithelial‑mesenchymal transition of lung adenocarcinoma". Oncology Reports 55, no. 4 (2026): 68. https://doi.org/10.3892/or.2026.9073
Copy and paste a formatted citation
x
Spandidos Publications style
Guo M, Li H, Zhao Z, Wang Y, Tang J, Zhong B and Zhao Y: Suppressive role of SCN4B in the epithelial‑mesenchymal transition of lung adenocarcinoma. Oncol Rep 55: 68, 2026.
APA
Guo, M., Li, H., Zhao, Z., Wang, Y., Tang, J., Zhong, B., & Zhao, Y. (2026). Suppressive role of SCN4B in the epithelial‑mesenchymal transition of lung adenocarcinoma. Oncology Reports, 55, 68. https://doi.org/10.3892/or.2026.9073
MLA
Guo, M., Li, H., Zhao, Z., Wang, Y., Tang, J., Zhong, B., Zhao, Y."Suppressive role of SCN4B in the epithelial‑mesenchymal transition of lung adenocarcinoma". Oncology Reports 55.4 (2026): 68.
Chicago
Guo, M., Li, H., Zhao, Z., Wang, Y., Tang, J., Zhong, B., Zhao, Y."Suppressive role of SCN4B in the epithelial‑mesenchymal transition of lung adenocarcinoma". Oncology Reports 55, no. 4 (2026): 68. https://doi.org/10.3892/or.2026.9073
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