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Visfatin facilitates esophageal cancer migration by suppressing miR‑3613‑5p expression and promoting VEZF1/VCAN production

  • Authors:
    • Chang-Lun Huang
    • Shubham Suresh Ghule
    • Yu-Hsiang Chang
    • Hsiao-Chi Tsai
    • Ming-Yu Lien
    • Jeng-Hung Guo
    • Chun-Lin Liu
    • Po-I Liu
    • Chih-Hsin Tang
  • View Affiliations / Copyright

    Affiliations: Division of General Thoracic Surgery, Department of Surgery, Changhua Christian Hospital, Changhua, Changhua 500, Taiwan, R.O.C., Graduate Institute of Biomedical Science, China Medical University, Taichung 404328, Taiwan, R.O.C., Program for Cancer Biology and Drug Discovery, China Medical University, Taichung 404328, Taiwan, R.O.C., Department of Medicine Research, China Medical University Beigang Hospital, Beigang, Yunlin 651012, Taiwan, R.O.C., School of Medicine, China Medical University, Taichung 404328, Taiwan, R.O.C., Department of Neurosurgery, China Medical University Hospital, Taichung 404328, Taiwan, R.O.C., Department of General Thoracic Surgery, Asia University Hospital, Taichung 413, Taiwan, R.O.C., Department of Pharmacology, School of Medicine, China Medical University, Taichung 404328, Taiwan, R.O.C.
    Copyright: © Huang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 128
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    Published online on: July 29, 2025
       https://doi.org/10.3892/or.2025.8961
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Abstract

Esophageal cancer, one of the most prevalent types of cancer worldwide, frequently exhibits distant metastases. The adipokine visfatin is implicated in cancer progression and metastasis. However, the mechanisms by which visfatin regulates motility in esophageal cancer remain unclear. Bioinformatics analysis showed levels of visfatin were higher in patients with metastatic esophageal cancer than in those with primary esophageal cancer. Cell motility assay revealed that visfatin stimulation enhanced the migration and invasion of esophageal cancer cells. Treatment with or without visfatin (30 ng/ml) in KYSE410 cells followed by miRNA sequencing, revealed that miR‑3613‑5p controlled visfatin‑induced cell motility. Further cell migration, invasion, qPCR and western blot assay shows that visfatin promoted esophageal cancer cell migration by decreasing miR‑3613‑5p expression and subsequently increasing vascular endothelial zinc finger 1/versican production. Thus, the visfatin/miR‑3613‑5p axis may be a promising target for inhibiting esophageal cancer cell migration and invasion.
View Figures

Figure 1

Visfatin promotes the migration and
invasion of esophageal cancer cells. (A) Visfatin mRNA levels in
normal, primary and metastatic esophageal cancer tissues from TCGA
dataset (UALCAN). Expression increased with lymph node metastasis
(N0-N1), with significant upregulation in metastatic tissue
(P=0.0194). Transwell (B) migration and (C) invasion assays in
KYSE410 cells treated with visfatin. Crystal violet-stained images
and quantification show dose-dependent increases in cell migration
and invasion. *P<0.05 vs. Control. TCGA, The Cancer Genome
Atlas; UALCAN, The University of Alabama at Birmingham Cancer data
analysis Portal; ns, not significant.

Figure 2

Visfatin enhances cell migration by
inhibiting mir-3613-5p. (A) Heatmap and (B) volcano plot of
differentially expressed mirs in KYSE-410 cells treated with
visfatin. Cells were (C) stimulated with visfatin and (D)
transfected with mir-3613-5p mimic and mir expression was verified
by reverse transcription-quantitative PCR. (E) KYSE410 cells
transfected with miR-3613-5p mimic with or without visfatin
treatment and cell migration and invasion was checked. (F)
quantification of cell migration and cell invasion assay. (G) CE81T
cells were transfected with mir-3613-5p mimic and treated with or
without visfatin; cell migration and invasion were examined. (H)
quantification of cell migration and cell invasion assay.
*P<0.05 vs. Control; #P<0.05 vs. Visfatin. Mir,
microRNA.

Figure 3

VEZF1, regulated by miR-3613-5p, is
involved in visfatin-induced esophageal cancer cell migration. (A)
miR databases (TargetScan, miRTarBase, miRDB and ENCORI) predicted
that miR-3613-5p targets seven potential candidates. (B) Gene
levels in patients with esophageal cancer retrieved from TCGA. (C)
VEZF1 gene levels in normal tissue and tissues from patients with
primary and metastatic esophageal cancer retrieved from TCGA. (D)
KYSE 410 cells were stimulated with visfatin, and VEZF1 expression
was examined using reverse transcription-quantitative PCR and (E)
western blot analysis. (F) VEZF1 siRNA transfection efficiency
examined by western blot. (G) KYSE410 cells were transfected with
VEZF1 siRNA and treated with visfatin, then cell (H) migration and
(I) cell invasion was examined. (J) CE81T cells were transfected
with VEZF1 siRNA followed by visfatin treatment and then (K)
Migration and (L) invasion was examined. *P<0.05 vs. control;
#P<0.05 vs. visfatin. miR, microRNA; TCGA, The Cancer
Genome Atlas; VEZF1, Vascular endothelial Zinc Finger 1; si, small
interfering; FC, fold change; ANP32B, Acidic Leucine-Rich Nuclear
Phosphoprotein 32 Family Member B; CDK, Cyclin-dependent kinase;
F11R, F11 receptor; LMNB2, Lamin B2; FLVCR, Feline Leukemia Virus
subgroup C Receptor 1; QSER, Glutamine and Serine-rich protein 1;
ns, not significant.

Figure 4

miR-3613-5p directly binds the 3′-UTR
of the VEZF1 gene. (A) Binding site of miR-3613-5p on the VEZF1
3′-UTR. (B) Cells were transfected with luciferase plasmids and
miR-3613-5p mimic, and luciferase activity was examined. Cells were
transfected with a miR-3613-5p mimic and treated with visfatin;
VEZF1 expression was examined using (C) reverse
transcription-quantitative PCR and (D) western blot assay.
*P<0.05 vs. control; #P<0.05 vs. visfatin. miR,
microRNA; UTR, untranslated region; VEZF1, Vascular Endothelial
Zinc Finger 1; wt, wild Type; mt, mutant; ns, not significant.

Figure 5

VCAN is highly expressed in patients
with esophageal cancer. (A) Spearman correlation analysis from
TCGA. (B) A total of 13 genes correlated with VEZF1 and were
associated with cell adhesion functions. (C) Gene levels in
patients with esophageal cancer retrieved from the GSE161533 and
TCGA database. (D) Kaplan-Meier analysis of VCAN level in normal
and cancer stage in ESCC *P<0.05. VCAN, Versican; VEZF1,
Vascular Endothelial Zinc Finger 1; TCGA, The Cancer Genome Atlas;
FC, Fold Change; PLXNC1, Plexin C1; ERBIN, interacting protein;
CLDN, claudin; ARAHGAP, Rho GTPase-activating protein 5; RGMB,
Repulsive Guidance Molecule BMP Co-Receptor B; ITGBL, Integrin
subunit beta 1; PTPRK, Protein Tyrosine Phosphatase Receptor Type
Kappa; DDR, DNA Damage Response; PARD, Par-3 family cell polarity
regulator; SPECC1L, sperm antigen with calponin homology and
coiled-coil domains 1 like; CDON, Cell Adhesion Associated,
Oncogene Regulated; PTPRT, protein tyrosine phosphatase, receptor
type, T.

Figure 6

VCAN is involved in visfatin-induced
esophageal cancer migration. Cells were stimulated with visfatin
and VCAN expression was examined using (A) RT-qPCR and (B) western
blot analysis. (C) VCAN siRNA transfection efficiency confirmed by
western blot analysis. (D) KYSE410 cells were transfected with or
without VCAN siRNA followed by visfatin treatment and cell (E)
migration and (F) invasion were examined. (G) CE81T cells were
transfected with VCAN siRNA and treated with visfatin; cell (H)
migration and (I) invasion were examined. Cells were transfected
with miR-3613-5p mimic or VEZF-1 siRNA and treated with visfatin;
VCAN expression was examined using (J) RT-qPCR and (K) western
blotting. (L) KYSE 410 cells treated with PI3K (Ly294002), AKT and
mTOR (rapamycin) inhibitors and visfatin treatment to assess
effects on (M) migration and (N) invasion. (O) CE81T cells treated
with PI3K (Ly294002), AKT and mTOR (rapamycin) inhibitors and
visfatin treatment were assayed for (P) cell migration and (Q)
invasion. Gene Expression Omnibus dataset GSE77861 shows
significantly increased mRNA expression of (R) NAMPT, (S)
miR-3613-5p, (T) VEZF1 and (U) VCAN in esophageal cancer compared
with normal tissue. *P<0.05 vs. control; #P<0.05
vs. visfatin. miR, microRNA; VCAN, versican; RT-q, Reverse
Transcriptase Quantitative; si, small interfering; VEZf-1, Vascular
Endothelial Zinc Finger 1; NAMPT, Nicotinamide
phosphoribosyltransferase; Akti, Akt inhibitor.

Figure 7

Mechanisms underlying the roles of
visfatin in esophageal cancer cell migration and invasion. Visfatin
enhances cell migration and invasion in esophageal cancer cells.
The inhibition of miR-3613-5p and the promotion of the VEZF1/VCAN
axis mediate visfatin-induced esophageal cancer cell motility. miR,
microRNA; VEZF1, Vascular Endothelial Zinc Finger 1; VCAN,
Versican; UTR, Untranslated region.
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Copy and paste a formatted citation
Spandidos Publications style
Huang C, Ghule SS, Chang Y, Tsai H, Lien M, Guo J, Liu C, Liu P and Tang C: Visfatin facilitates esophageal cancer migration by suppressing miR‑3613‑5p expression and promoting VEZF1/VCAN production. Oncol Rep 54: 128, 2025.
APA
Huang, C., Ghule, S.S., Chang, Y., Tsai, H., Lien, M., Guo, J. ... Tang, C. (2025). Visfatin facilitates esophageal cancer migration by suppressing miR‑3613‑5p expression and promoting VEZF1/VCAN production. Oncology Reports, 54, 128. https://doi.org/10.3892/or.2025.8961
MLA
Huang, C., Ghule, S. S., Chang, Y., Tsai, H., Lien, M., Guo, J., Liu, C., Liu, P., Tang, C."Visfatin facilitates esophageal cancer migration by suppressing miR‑3613‑5p expression and promoting VEZF1/VCAN production". Oncology Reports 54.4 (2025): 128.
Chicago
Huang, C., Ghule, S. S., Chang, Y., Tsai, H., Lien, M., Guo, J., Liu, C., Liu, P., Tang, C."Visfatin facilitates esophageal cancer migration by suppressing miR‑3613‑5p expression and promoting VEZF1/VCAN production". Oncology Reports 54, no. 4 (2025): 128. https://doi.org/10.3892/or.2025.8961
Copy and paste a formatted citation
x
Spandidos Publications style
Huang C, Ghule SS, Chang Y, Tsai H, Lien M, Guo J, Liu C, Liu P and Tang C: Visfatin facilitates esophageal cancer migration by suppressing miR‑3613‑5p expression and promoting VEZF1/VCAN production. Oncol Rep 54: 128, 2025.
APA
Huang, C., Ghule, S.S., Chang, Y., Tsai, H., Lien, M., Guo, J. ... Tang, C. (2025). Visfatin facilitates esophageal cancer migration by suppressing miR‑3613‑5p expression and promoting VEZF1/VCAN production. Oncology Reports, 54, 128. https://doi.org/10.3892/or.2025.8961
MLA
Huang, C., Ghule, S. S., Chang, Y., Tsai, H., Lien, M., Guo, J., Liu, C., Liu, P., Tang, C."Visfatin facilitates esophageal cancer migration by suppressing miR‑3613‑5p expression and promoting VEZF1/VCAN production". Oncology Reports 54.4 (2025): 128.
Chicago
Huang, C., Ghule, S. S., Chang, Y., Tsai, H., Lien, M., Guo, J., Liu, C., Liu, P., Tang, C."Visfatin facilitates esophageal cancer migration by suppressing miR‑3613‑5p expression and promoting VEZF1/VCAN production". Oncology Reports 54, no. 4 (2025): 128. https://doi.org/10.3892/or.2025.8961
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