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Zinc finger protein 514 promotes esophageal cancer progression by enhancing cell proliferation, migration and invasion

  • Authors:
    • Lin Lv
    • Xiaoming Sun
    • Houlu Zhang
    • Guangxu Wang
    • Chao Zhang
    • Haibo Liu
    • Liangming Zhu
  • View Affiliations / Copyright

    Affiliations: Department of Thoracic Surgery, Jinan Central Hospital, Shandong University, Jinan, Shandong 250013, P.R. China, School of Clinical Medicine, Shandong Second Medical University, Weifang, Shandong 261000, P.R. China, Graduate Department, Shandong First Medical University, Jinan, Shandong 250000, P.R. China
    Copyright: © Lv et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 14
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    Published online on: October 24, 2025
       https://doi.org/10.3892/mmr.2025.13724
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Abstract

Esophageal cancer (EC), a malignant tumor occurring in the upper gastrointestinal tract, is the seventh most common cancer worldwide. Zinc finger proteins (ZNFs), the most abundant family of transcription factors in humans, serve an important role in the initiation and progression of various malignant tumors. However, the function of ZNFs in EC remains unclear. The present study aimed to elucidate the role of ZNF514 in the development and progression of EC and to investigate its underlying mechanism. The mRNA and protein expression levels of ZNF514 were assessed using reverse transcription‑quantitative PCR and western blotting. To assess functional roles, multiple cellular assays were performed, including 5‑ethynyl‑2'‑deoxyuridine incorporation, Cell Counting Kit‑8, wound healing, colony formation and Transwell assays. Subsequently, for transcriptomics analysis, Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses, Gene Set Enrichment Analysis and Ingenuity Pathway Analysis (IPA) were performed. In EC, ZNF514 exhibited high expression at both the mRNA and protein levels. Additionally, ZNF514 influenced the migration, invasion and proliferation of EC cells. Gene enrichment analyses and IPA demonstrated that ZNF514 knockdown significantly affected multiple signaling pathways, such as Fcγ receptors, the complement system, G‑protein coupled receptors (GPCRs)‑related receptors, the ribosomal S6 kinase (RSK) pathway, Ras/MEK, PI3K/AKT, STAT3, nucleotide‑binding oligomerization domain‑containing protein (NOD) and NF‑κB pathways. In conclusion, the present study indicated that the anticancer mechanisms induced by ZNF514 knockdown may be related to the enhancement of Fcγ receptor and complement system activation, as well as the inhibition of GPCR, RSK, Ras/MEK, PI3K/AKT, STAT3, NOD1/2 and NF‑κB pathways.
View Figures

Figure 1

Figure 2

Figure 3

ZNF514 is significantly upregulated
in EC tissues and cells. (A) Analysis of RNA expression differences
of ZNF514 in normal and cancer tissues across various types of
cancer using the Tumor Immune Estimation Resource database. (B) RNA
expression differences between H and paired cancer tissues in
patients with EC. (C) Gene alteration frequency of ZNF514 in EC.
(D) Western blot analysis of ZNF514 protein expression differences
in Het-1a and EC cell lines. (E) Western blot analysis of ZNF514
protein expression differences in the tumors and adjacent tissues
of patients with EC. Semi-quantitative analyses of ZNF514 protein
expression in (F) Het-1a and EC cell lines, and (G) tumor and
adjacent tissues of patients with EC normalized to GAPDH. (H)
Reverse transcription-quantitative PCR analysis of ZNF514
expression levels in Het-1a and EC cell lines. (I) Representative
images of immunohistochemical staining for ZNF514 in the tumors and
adjacent tissues of patients with EC. Scale bar, 1000 µm.
*P<0.05, **P<0.01, ***P<0.001 and ****P<0.0001. H,
Healthy; T, tumor; ZNF514, zinc finger protein 514; EC, esophageal
cancer; ns, not significant; CNA, copy number alteration; TCGA, The
Cancer Genome Atlas.

Figure 4

Downregulation of ZNF514 inhibits
proliferation, migration and invasion of Kyse-150 cells. ZNF514
mRNA and protein expression levels in Kyse-150 cells transfected
with targeted siRNAs were analyzed using (A) reverse
transcription-quantitative PCR and (B) western blot analysis,
alongside (C) semi-quantitative analysis of ZNF514 protein levels.
(D) Effects of ZNF514 knockdown on the migration and invasion of
Kyse-150 cells was assessed using Transwell assays; (E)
quantitative analysis of the migration assay, (F) quantitative
analysis of the invasion assay. Scale bar, 100 µm. (G) Wound
healing assays were conducted to evaluate the effect of ZNF514
knockdown on the migration of Kyse-150 cells; (H) quantitative
analysis of cell migration rate. Scale bar, 200 µm. (I)
Quantitative analysis of colony formation, (J) colony formation,
(K) Cell Counting Kit −8, (L) EdU assays, and (M) quantitative
analysis of the EdU assay were utilized to evaluate the effects of
ZNF514 knockdown on the proliferation of Kyse-150 cells. Scale bar,
100 µm. *P<0.05, **P<0.01, ***P<0.001 and ****P<0.0001
vs. si-NT. NT, non-targeting; ZNF514, zinc finger protein 514;
CCK-8, Cell Counting Kit-8; si, small interfering; EdU,
5-ethynyl-2′-deoxyuridine.

Figure 5

OE of ZNF514 promotes proliferation,
migration and invasion of Kyse-510 cells. Analysis of ZNF514 mRNA
and protein expression in Kyse-510 cells transfected with targeted
OE plasmid using (A) reverse transcription-quantitative PCR and (B)
western blot analysis, along with (C) semi-quantification of ZNF514
protein. (D) Transwell assays were performed to evaluate the
effects of ZNF514 OE on the migration and invasion of Kyse-510
cells; (E) quantitative analysis of the migration assay, (F)
quantitative analysis of the invasion assay. Scale bar, 200 µm. (G)
Wound healing assays were conducted to assess the impact of ZNF514
OE on the migration of Kyse-510 cells. (H) quantitative analysis of
cell migration rate. Scale bar, 100 µm. (I) Quantitative analysis
of colony formation, (J) Colony formation, (K) Cell Counting Kit-8,
(L) EdU assays, and (M) quantitative analysis of the EdU assay were
utilized to evaluate the effects of ZNF514 OE on the proliferation
of Kyse-510 cells. Scale bar, 100 µm. *P<0.05, **P<0.01,
***P<0.001 and ****P<0.0001 vs. vector. ZNF514, zinc finger
protein 514; OE, overexpression; EdU,
5-ethynyl-2′-deoxyuridine.

Figure 6

RNA sequencing analysis of esophageal
cancer cells reveals the mechanisms by which downregulation of
ZNF514 contributes to cancer progression. (A) Bar graph displaying
the number of genes affected by ZNF514 knockdown, selected based on
the criteria of |log2(Fold-change)|>1 and q-value
<0.05. (B) Volcano plot illustrating the expression of DEGs,
with red indicating upregulated genes and blue signifying
downregulated genes. (C) Scatter plot showing the correlation of
DEGs, with red indicating upregulated genes and blue signifying
downregulated genes. (D) GO functional classification of DEGs, (E)
GO enrichment analysis of DEGs, (F) KEGG pathway classification of
DEGs, and (G) KEGG pathway enrichment analysis of DEGs. (H)
Protein-protein interaction analysis was conducted based on the
DEGs. DEG, differentially expressed gene; ZNF514, zinc finger
protein 514; GO, Gene Ontology; KEGG, Kyoto Encyclopedia of Genes
and Genomes; si-NT, siRNA-non-targeting.

Figure 7

GSEA of differentially expressed
genes. The enrichment plots demonstrated that (A) FCGR-dependent
phagocytosis, (B) FCGR activation and (C) initial triggering of
complement were activated, whereas (D) GPCR ligand binding, (E) Gβγ
signaling through PI3Kγ and (F) RSK activation were inhibited;
FCGR, Fcγ receptor; GPCR, G-protein coupled receptor; RSK,
ribosomal S6 kinase.

Figure 8

Ingenuity Pathway Analysis of DEGs
from RNA sequencing analysis. The top 10 significantly (A)
upregulated and (B) downregulated genes following ZNF514 knockdown
in Kyse-150 cells, based on log2(Fold-change). (C)
Enrichment analysis of disease-related genes. (D) Pathway
enrichment analysis based on DEGs. The pathways are indicated in
orange for significant activation and blue for significant
inhibition, based on z-score analysis. DEG, differentially
expressed gene; ZNF514, zinc finger protein 514.

Figure 9

IPA shows that ZNF514 knockdown
affects multiple cancer-related pathways. (A) The affected PI3K/AKT
pathway, and its upstream and downstream relationships in the
Molecular Mechanisms in Cancer pathway. (B) Activation of TBX3, and
(C) inhibition of TLR5, with the downstream molecules influenced by
these factors shown. Solid line, direct interaction; dashed line,
indirect interaction. Arrow, activation; Blocked arrow, inhibition.
Blue, leads to inhibition; Orange, leads to activation; Yellow,
findings inconsistent with state of downstream molecule; Gray,
effect not predicted.
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Copy and paste a formatted citation
Spandidos Publications style
Lv L, Sun X, Zhang H, Wang G, Zhang C, Liu H and Zhu L: Zinc finger protein 514 promotes esophageal cancer progression by enhancing cell proliferation, migration and invasion. Mol Med Rep 33: 14, 2026.
APA
Lv, L., Sun, X., Zhang, H., Wang, G., Zhang, C., Liu, H., & Zhu, L. (2026). Zinc finger protein 514 promotes esophageal cancer progression by enhancing cell proliferation, migration and invasion. Molecular Medicine Reports, 33, 14. https://doi.org/10.3892/mmr.2025.13724
MLA
Lv, L., Sun, X., Zhang, H., Wang, G., Zhang, C., Liu, H., Zhu, L."Zinc finger protein 514 promotes esophageal cancer progression by enhancing cell proliferation, migration and invasion". Molecular Medicine Reports 33.1 (2026): 14.
Chicago
Lv, L., Sun, X., Zhang, H., Wang, G., Zhang, C., Liu, H., Zhu, L."Zinc finger protein 514 promotes esophageal cancer progression by enhancing cell proliferation, migration and invasion". Molecular Medicine Reports 33, no. 1 (2026): 14. https://doi.org/10.3892/mmr.2025.13724
Copy and paste a formatted citation
x
Spandidos Publications style
Lv L, Sun X, Zhang H, Wang G, Zhang C, Liu H and Zhu L: Zinc finger protein 514 promotes esophageal cancer progression by enhancing cell proliferation, migration and invasion. Mol Med Rep 33: 14, 2026.
APA
Lv, L., Sun, X., Zhang, H., Wang, G., Zhang, C., Liu, H., & Zhu, L. (2026). Zinc finger protein 514 promotes esophageal cancer progression by enhancing cell proliferation, migration and invasion. Molecular Medicine Reports, 33, 14. https://doi.org/10.3892/mmr.2025.13724
MLA
Lv, L., Sun, X., Zhang, H., Wang, G., Zhang, C., Liu, H., Zhu, L."Zinc finger protein 514 promotes esophageal cancer progression by enhancing cell proliferation, migration and invasion". Molecular Medicine Reports 33.1 (2026): 14.
Chicago
Lv, L., Sun, X., Zhang, H., Wang, G., Zhang, C., Liu, H., Zhu, L."Zinc finger protein 514 promotes esophageal cancer progression by enhancing cell proliferation, migration and invasion". Molecular Medicine Reports 33, no. 1 (2026): 14. https://doi.org/10.3892/mmr.2025.13724
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