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Overexpression of ATP1B2 promotes cancer cell migration and inhibits apoptosis in patients with esophageal squamous cell carcinoma

  • Authors:
    • Fang-Fei Liu
    • Hui Wen
    • Xiao-Bo Liu
    • Sheng-Bao Li
    • Shu Jin
    • Zi-Ye Gao
    • Qiang Tong
  • View Affiliations / Copyright

    Affiliations: Department of Intensive Care Unit, Yuebei People's Hospital, Shaoguan, Guangdong 512000, P.R. China, Department of Gastroenterology, Taihe Hospital, Hubei University of Medicine, Shiyan, Hubei 442000, P.R. China, Department of Oncology, Taihe Hospital, Hubei University of Medicine, Shiyan, Hubei 442000, P.R. China
    Copyright: © Liu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 96
    |
    Published online on: June 17, 2025
       https://doi.org/10.3892/or.2025.8929
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Abstract

The present study aimed to investigate the expression of ATP1B2 in esophageal squamous cell carcinoma (ESCC) and its biological effects. A total of 44 patients with ESCC who underwent surgical resection at Taihe Hospital between December 1, 2017 and December 1, 2018 were enrolled. The expression levels of ATP1B2 in cancerous and adjacent normal tissues were assessed. The present study also examined the associations between ATP1B2 expression and clinicopathological features and patient prognosis. The influence of ATP1B2 on ESCC cell proliferation, migration, cell cycle progression and apoptosis was evaluated using the methylcyclopentadienyl manganese tricarbonyl assay, plate cloning, scratch assay and flow cytometry. Furthermore, the effects of ouabain on these cellular processes were investigated. The results demonstrated that patients with high ATP1B2 expression exhibited significantly shorter overall survival than did those with low ATP1B2 expression (37.3 months vs. 43.1 months; Z=7.52; P<0.05). ATP1B2 expression, tumor invasion and lymph node metastasis were significantly associated (P<0.05). Notably, the overexpression of ATP1B2 correlated with reduced survival rates. ATP1B2 knockdown hindered cell migration and induced apoptosis, whereas ATP1B2 overexpression facilitated migration and impeded apoptosis. Ouabain treatment suppressed proliferation and migration in cells overexpressing ATP1B2 and caused cell cycle arrest in the G1/S phase. In conclusion, ATP1B2 overexpression is associated with poor prognosis in patients with ESCC by enhancing cancer cell migration and reducing apoptosis. Ouabain is a potential targeted therapeutic agent for ESCC.
View Figures

Figure 1

ATP1B2 expression in esophageal
cancer and adjacent tissues. (A) In the tissues of ESCC, ATP1B2
protein is mainly located in the nucleus, stained brown, and
partially located in the cytoplasm, stained light yellow. (B)
Expression of ATP1B2 protein in tissues adjacent to ESCC. (C)
Kaplan-Meier method was used to analyze the survival of patients
with ESCC with different levels of ATP1B2 expression. The survival
rate of the high expression group was significantly lower than that
of the low expression group. (D) The expression level of ATP1B2 in
immortalized ESCC cell lines was significantly higher in EC109
cells than in Het-1A. (E) In esophageal cancer EC109, the
expression of ATP1B2 mRNA in ATP1B2-shRNA group was significantly
lower than that of non-Sil shRNA. (F) In esophageal cancer KYSE150
cells, the expression of ATP1B2 mRNA in ATP1B2-over expression
cells was significantly higher than that in control group. (G)
Western blotting detected the overexpression and silencing of
ATP1B2 protein expression. (H and I) Densitometric analysis of
western blotting of (H) silencing of ATP1B2 protein and (I)
overexpression of ATP1B2 protein. #P<0.05 and
##P<0.01. ESCC, esophageal squamous cell carcinoma;
shRNA, short hairpin RNA.

Figure 2

Effect of ATP1B2 on biological
behavior of esophageal squamous cell carcinoma cells: invasion,
migration ability and cell cycle. (A and B) Observation of cell
scratch healing in knockdown and overexpression ATP1B2 and
empty-vector control group under fluorescence microscopy. (C and D)
Histogram of cell scratch healing in knockdown and overexpression
ATP1B2 and empty-vector control group. (E and F) Cell cycle
distribution of overexpression ATP1B2 and empty-vector control
group. (G) Histogram of cell cycle of ATP1B2 overexpression and
empty-vector control group. (H and I) Cell cycle distribution of
ATP1B2 knockdown and empty-vector control group. (J) Histogram of
cell cycle of ATP1B2 knockdown and empty-vector control group.
shRNA, short hairpin RNA. #P<0.05.

Figure 3

Effect of ATP1B2 on biological
behavior of esophageal cancer cells: Proliferation, invasion,
migration ability, cell cycle and apoptosis. (A and B) Comparison
of apoptosis between the experimental group and the empty-vector
control group with ATP1B2 knockdown in ESCC cell line EC109 cells.
(C and D) Comparison of apoptosis between the experimental group
with overexpression of ATP1B2 and the empty-vector control group in
ESCC cell line KYSE150 cells. (E) EC109 cell apoptosis was higher
than that of control group after ATP1B2 knockout. (F) After
overexpression of ATP1B2, the apoptosis of KYSE150 cells was lower
than that of the control group. #P<0.05. ESCC,
esophageal squamous cell carcinoma; shRNA, short hairpin RNA.

Figure 4

Effect of ATP1B2 on biological
behavior of esophageal cancer cells: Proliferation. (A and B) The
research results in the human ESCC cell line EC109. (A) Knockdown
of the ATP1B2 experimental group; (B) empty-vector control group.
The results showed that knocking down ATP1B2 can promote the
proliferation of EC109 cells. (C and D) The study in the human ESCC
line KYSE150. (C) Experimental group overexpressing ATP1B2; (D)
control group. The results revealed that overexpressing ATP1B2 can
inhibit the proliferation of KYSE150 cells. (E) Plate cloning
experiment demonstrated that knockdown of ATP1B2 gene promoted the
proliferation of EC109 cells. (F) Plate cloning experiment showed
that overexpression of ATP1B2 inhibited the proliferation of
KYSE150 cells. (G) MTT identified that knockdown of ATP1B2 gene
promoted the proliferation of EC109 cells. (H) MTT showed that
overexpression of ATP1B2 inhibited the proliferation of KYSE150
cells. #P<0.05 and **P<0.01. ESCC, esophageal
squamous cell carcinoma; shRNA, short hairpin RNA.

Figure 5

RT-qPCR determination of expression
content of epithelial-mesenchymal transformation-related genes.
(A-D) RT-qPCR detection in the experimental group overexpressed
with ATP1B2 and the empty-vector control group of (A) MMP7; (B)
MMP10 (C) RAC1 and (D) P53. #P<0.05. RT-qPCR, reverse
transcription-quantitative PCR.

Figure 6

Expression of epithelial-mesenchymal
transformation-related proteins was detected by western blotting.
(A) Western blotting of various proteins in EC109 cells and
EC109-ATP1B2-shRNA cells. (B-D) Densitometric analysis of
expression levels in EC109 cells and EC109-ATP1B2-shRNA cells of
(B) N-cadherin/β-actin; (C) E-cadherin/β-actin and (D) P53/β-actin
#P<0.05. shRNA, short hairpin RNA.

Figure 7

Protein expression levels of P53 and
RAC1 in ATP1B2-overexpressing cells. (A) Western blotting of P53
and RAC1 proteins in KYSE150 cells and EC109-ATP1B2-overexpressing
cells. (B and C) Densitometric analysis in KYSE150 cells and
EC109-ATP1B2-overexpressing cells of expression levels of (B)
P53/β-actin and (C) RAC1/β-actin.

Figure 8

Effect of different concentrations of
ouabain on proliferation of esophageal cancer cells. MTT test
confirmed that compared with the empty-vector control group, the
inhibitory effect of ouabain on the proliferation of esophageal
cancer cells overexpressing ATP1B2 was significantly enhanced with
the prolongation of the concentration and time of action. (A) MTT
detection of cancer cell proliferation in the empty-vector control
group. (B-D) Compared with the empty-vector control group, the
inhibitory effect on the proliferation of esophageal cancer cells
in the overexpressing ATP1B2 group of (B) 20 µl/l ouabain; (C) of
40 µl/l ouabain and (D) 60 µl/l ouabain. #P<0.05 and
**P<0.01. MTT, methylcyclopentadienyl manganese tricarbonyl.

Figure 9

Effect of ouabain on the migration
ability and cell cycle of esophageal cancer cells after
overexpression of ATP1B2. (A) Scratch test showed that the
inhibitory effect of ouabain on the migration ability of
overexpression ATP1B2 cells was significantly enhanced by adding 60
µl/l of ouabain. (B) Compared with the empty-vector control group,
the inhibitory effect of ouabain on the proliferation of esophageal
cancer cells overexpressing ATP1B2 was significantly enhanced with
the prolongation of the action concentration and time. (C and D)
The cell cycle experiment revealed that ouabain significantly
enhanced the inhibition of cell cycle arrest in G1/S
phase of esophageal cancer cells with overexpression of ATP1B2; (C)
experimental group over-expressing ATP1B2 with ouabain; (D) control
group with ouabain. (E) Inhibitory effect of E ouabain on cell
cycle arrest in G1/S phase of esophageal cancer cells
overexpressing ATP1B2 was significantly enhanced. **P<0.01.
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Spandidos Publications style
Liu F, Wen H, Liu X, Li S, Jin S, Gao Z and Tong Q: Overexpression of ATP1B2 promotes cancer cell migration and inhibits apoptosis in patients with esophageal squamous cell carcinoma. Oncol Rep 54: 96, 2025.
APA
Liu, F., Wen, H., Liu, X., Li, S., Jin, S., Gao, Z., & Tong, Q. (2025). Overexpression of ATP1B2 promotes cancer cell migration and inhibits apoptosis in patients with esophageal squamous cell carcinoma. Oncology Reports, 54, 96. https://doi.org/10.3892/or.2025.8929
MLA
Liu, F., Wen, H., Liu, X., Li, S., Jin, S., Gao, Z., Tong, Q."Overexpression of ATP1B2 promotes cancer cell migration and inhibits apoptosis in patients with esophageal squamous cell carcinoma". Oncology Reports 54.2 (2025): 96.
Chicago
Liu, F., Wen, H., Liu, X., Li, S., Jin, S., Gao, Z., Tong, Q."Overexpression of ATP1B2 promotes cancer cell migration and inhibits apoptosis in patients with esophageal squamous cell carcinoma". Oncology Reports 54, no. 2 (2025): 96. https://doi.org/10.3892/or.2025.8929
Copy and paste a formatted citation
x
Spandidos Publications style
Liu F, Wen H, Liu X, Li S, Jin S, Gao Z and Tong Q: Overexpression of ATP1B2 promotes cancer cell migration and inhibits apoptosis in patients with esophageal squamous cell carcinoma. Oncol Rep 54: 96, 2025.
APA
Liu, F., Wen, H., Liu, X., Li, S., Jin, S., Gao, Z., & Tong, Q. (2025). Overexpression of ATP1B2 promotes cancer cell migration and inhibits apoptosis in patients with esophageal squamous cell carcinoma. Oncology Reports, 54, 96. https://doi.org/10.3892/or.2025.8929
MLA
Liu, F., Wen, H., Liu, X., Li, S., Jin, S., Gao, Z., Tong, Q."Overexpression of ATP1B2 promotes cancer cell migration and inhibits apoptosis in patients with esophageal squamous cell carcinoma". Oncology Reports 54.2 (2025): 96.
Chicago
Liu, F., Wen, H., Liu, X., Li, S., Jin, S., Gao, Z., Tong, Q."Overexpression of ATP1B2 promotes cancer cell migration and inhibits apoptosis in patients with esophageal squamous cell carcinoma". Oncology Reports 54, no. 2 (2025): 96. https://doi.org/10.3892/or.2025.8929
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