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Exosomal‑miR‑32‑5p directly targets FOXN2 to regulate the proliferation, migration and apoptosis of uterine corpus endometrial carcinoma via the PI3K/AKT/BCL‑2 pathway

  • Authors:
    • Xin Chen
    • Huihui Li
    • Xiang Cheng
    • Yirong Cai
    • Guangyi Guo
  • View Affiliations / Copyright

    Affiliations: Department of Oncology, Renmin Hospital of Wuhan University, Wuhan, Hubei 430060, P.R. China, Department of Healthcare Center, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430022, P.R. China, Department of Oncology, Traditional Chinese Medicine Hospital of Xishui County, Huanggang, Hubei 438000, P.R. China
    Copyright: © Chen et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 219
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    Published online on: April 8, 2026
       https://doi.org/10.3892/ol.2026.15574
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Abstract

The transfer of microRNAs (miRNAs) between cells through exosomes is crucial in controlling the expression of various target genes in the recipient cells, giving exosomal miRNAs the capability to control the advancement of tumors. The present study aimed to explore the function of exosomal miR‑32‑5p in the in vitro progression of uterine corpus endometrial carcinoma (UCEC). Bioinformatics analyses were applied to identify potential miRNAs in UCEC. Forkhead Box N2 (FOXN2) was subsequently predicted as a putative target gene of miR‑32‑5p, followed by examination through a luciferase reporter assay. Exosomes derived from UCEC cells or plasma samples of patients with UCEC were examined. The biological functions of miR‑32‑5p or exosomal miR‑32‑5p were determined by cell phenotype experiments. Protein and mRNA expression in UCEC cells were assessed through western blotting and quantitative reverse transcription‑PCR, respectively. miR‑32‑5p demonstrated the most significant effects on survival probability, therefore it was selected for further investigation in in the present study. miR‑32‑5p was observed to be enriched in plasma exosomes and significantly upregulated in cancer tissues, as well as in cancer plasma exosomes. The present findings demonstrated that exosomal‑miR‑32‑5p could induce the proliferation, migration and suppress apoptosis of UCEC cells through regulation of the FOXN2/PI3K/AKT/Bcl‑2 pathway, thus exhibiting potential as therapeutic target against UCEC.
View Figures

Figure 1

miRNA screening. (A) The volcano plot
of DE-miRNAs (n=656) comparing normal and UCEC samples from TCGA
database. Green represents 402 miRNAs that were differentially
upregulated in UCEC samples; red represents 254 miRNAs that were
differentially downregulated. (B) The intersection of the 656
DE-miRNAs with the 49 DE-miRNAs analyzed by Zhou et al
(21), 26 DE-miRNAs were
identified. (C) From the univariate Cox risk regression analysis,
140 miRNAs were screened as prognostic miRNAs in patients with
UCEC. The selected 140 miRNAs were intersected with 26 DE-miRNAs,
from which 7 DE-miRNAs were obtained. (D) Multivariate Cox
regression analysis was used to determine the independent
prognostic factors of UCEC. (E) Survival probability analysis for
miR-15a-5p, miR-1180-3p, miR-32-5p and miR-1197. DE, differentially
expressed; UCEC, uterine corpus endometrial carcinoma; miRNA,
microRNA. AIC, Akaike Information Criterion.

Figure 2

miR-32-5p is overexpressed in
UCEC-Exo. (A) The morphology of plasma exosomes was visualized
using transmission electron microscopy. Scale bar, 100 µm. (B) The
size distribution and concentration of plasma exosomes were
assessed by nanoparticle tracking analysis. The X-axis represents
the diameter (nm) of exosomes, while the Y-axis represents the
concentration of exosomes. (C) The protein levels of
exosome-positive markers (CD9, CD81 and TSG101) and
exosome-negative marker (Calnexin) in plasma exosomes were
determined using western blotting. (D) The expression of miR-32-5p
in normal-Exo and UCEC-Exo was assessed using quantitative reverse
transcription-PCR. **P<0.01 vs. normal-Exo. UCEC, uterine corpus
endometrial carcinoma; miRNA, microRNA; Exo, exosome; TSG101, tumor
susceptibility gene 101.

Figure 3

Expression of miR-32-5p in exosomes
isolated from UCEC cell lines. (A) The morphology of UCEC cell
line-derived exosomes was visualized using transmission electron
microscopy. Scale bar, 100 µm. (B) The size distribution and
concentration of UCEC cell line-derived exosomes were assessed by
nanoparticle tracking analysis. The X-axis represents the diameter
(nm) of exosomes, while Y-axis represents the concentration of
exosomes. (C) The protein levels of exosome-positive markers (CD9,
CD81 and TSG101) and exosome-negative marker (Calnexin) in UCEC
cell line-derived exosomes were determined using western blotting.
(D) The expression of miR-32-5p in HEC-1-A-Exo and Ishikawa-Exo was
detected using quantitative reverse transcription-PCR.
***P<0.001 vs. HEC-1-A-Exo. UCEC, uterine corpus endometrial
carcinoma; miRNA, microRNA; Exo, exosome; TSG101, tumor
susceptibility gene 101.

Figure 4

Effects of miR-32-5p on the
proliferative capacities, migratory abilities and apoptotic rate of
UCEC cells. (A) The expression of miR-32-5p in HEC-1-A and Ishikawa
cells were then determined using quantitative reverse
transcription-PCR. ***P<0.001 vs. HEC-1-A cells. (B) The
expression of miR-32-5p in HEC-1-A cells after transfection of
miR-32-5p inhibitor or in Ishikawa cells after transfection of
miR-32-5p mimic. (C) The viability of UCEC cell lines were measured
using the CCK-8 assay. (D) Relative colony formation efficiency of
UCEC cell lines was evaluated using the colony formation assay. (E)
EdU-positive cells were measured through EdU proliferation assay.
(Scale bar, 100 µm). (F) The migratory potential of UCEC cell lines
were determined via Transwell migration assay. (Scale bar, 100 µm).
(G) Flow cytometry was used to detect apoptosis.
*P<0.05, **P<0.01, ***P<0.001 vs. miR-32-5p
inhibitor-NC or miR-32-5p mimic-NC. UCEC, uterine corpus
endometrial carcinoma; miRNA, microRNA; NC, negative control; EdU,
5-ethynyl-2′-deoxyuridine.

Figure 5

miR-32-5p negatively regulates FOXN2
and activates PI3K/AKT pathway. (A) The Starbase software shows the
potential binding site for miR-32-5p and FOXN2. (B) A dual
luciferase reporter assay was conducted to assess the relationship
between miR-32-5p and FOXN2. ***P<0.001 vs. FOXN2-WT
+ mimic-NC. (C) The mRNA expression of AKT, PI3K, Bcl-2 and FOXN2
in HEC-1-A cells transfected with miR-32-5p inhibitor or
inhibitor-NC was detected using qRT-PCR. *P<0.05, **P<0.01,
***P<0.001 vs. miR-32-5p inhibitor-NC. (D) The mRNA expression
of AKT, PI3K, Bcl-2 and FOXN2 in Ishikawa cells transfected with
miR-32-5p mimic or mimic-NC was detected by qRT-PCR. *P<0.05,
**P<0.01 and ***P<0.001 vs. miR-32-5p mimic-NC. (E) The
protein levels of AKT, PI3K, p-PI3K, Bcl-2, FOXN2 and the ratio of
p-AKT/AKT in HEC-1-A cells transfected with miR-32-5p inhibitor or
inhibitor-NC were measured using western blotting. *P<0.05 and
***P<0.001 vs. miR-32-5p inhibitor-NC. (F) The protein levels of
AKT, PI3K, p-PI3K, Bcl-2, FOXN2 and the ratio of p-AKT/AKT in
Ishikawa cells transfected with miR-32-5p mimic or mimic-NC were
measured using western blotting. *P<0.05 and ***P<0.001 vs.
miR-32-5p inhibitor-NC. UCEC, uterine corpus endometrial carcinoma;
miRNA, microRNA; NC, negative control; NS, not significant; WT,
wild-type; MUT, mutant; qRT-PCR, quantitative reverse
transcription-PCR; p-, phosphorylated; FOXN2, Forkhead Box N2.

Figure 6

Effects of miR-32-5p-mediated FOXN2
on PI3K/AKT/Bcl-2 pathway. (A) Following transfection of
OE-FOXN2/OE-NC in Ishikawa cells, the mRNA expression of FOXN2 was
determined via qRT-PCR. Following transfection of miR-32-5p-mimic
or OE-FOXN2 in Ishikawa cells, (B) the mRNA expression and (C)
protein levels of FOXN2/PI3K/AKT/Bcl-2 and the ratio of p-AKT/AKT
were determined through qRT-PCR and western blotting, respectively.
*P<0.05, **P<0.01, ***P<0.001, ****P<0.0001. UCEC,
uterine corpus endometrial carcinoma; miRNA, microRNA; NC, negative
control; NS, not significant; WT, wild-type; MUT, mutant; qRT-PCR,
quantitative reverse transcription-PCR; p-, phosphorylated; FOXN2,
Forkhead Box N2; OE, overexpression.

Figure 7

Exo-miR-32-5p regulates the
proliferation, migration and apoptosis of UCEC cells through
regulating FOXN2 expression and PI3K/AKT/Bcl-2 pathway. The impact
of Exo-miR-32-5p on the proliferation, migration and apoptosis of
UCEC cells were examined utilizing a co-culture model. (A)
Exo-miR-32-5p expression in HEC-1-A cells transfected with
miR-32-5p-inhibitor or in Ishikawa cells transfected with
miR-32-5p-mimic was determined through qRT-PCR. (B) The viability
of UCEC cell lines were measured using the CCK-8 assay. (C)
Relative colony formation efficiency of UCEC cell lines was
evaluated using the colony formation assay. (D) EdU-positive cells
were measured through EdU proliferation assay. (Scale bar, 100 µm).
(E) The migratory potential of UCEC cell lines were determined via
the Transwell migration assay. (F) Flow cytometry was used to
detect apoptosis. (G) The mRNA expression of AKT, PI3K, Bcl-2 and
FOXN2 was detected by qRT-PCR. (H) The protein levels of AKT, PI3K,
p-PI3K, Bcl-2, FOXN2 and the ratio of p-AKT/AKT were measured by
western blotting. **P<0.01, ***P<0.001 vs. Exo-miR-32-5p
inhibitor-NC or Exo-miR-32-5p mimic-NC. UCEC, uterine corpus
endometrial carcinoma; miRNA, microRNA; NC, negative control; NS,
not significant; WT, wild-type; MUT, mutant; qRT-PCR, quantitative
reverse transcription-PCR; p-, phosphorylated; FOXN2, Forkhead Box
N2; EdU, 5-ethynyl-2′-deoxyuridine.
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Copy and paste a formatted citation
Spandidos Publications style
Chen X, Li H, Cheng X, Cai Y and Guo G: Exosomal‑miR‑32‑5p directly targets FOXN2 to regulate the proliferation, migration and apoptosis of uterine corpus endometrial carcinoma via the PI3K/AKT/BCL‑2 pathway. Oncol Lett 31: 219, 2026.
APA
Chen, X., Li, H., Cheng, X., Cai, Y., & Guo, G. (2026). Exosomal‑miR‑32‑5p directly targets FOXN2 to regulate the proliferation, migration and apoptosis of uterine corpus endometrial carcinoma via the PI3K/AKT/BCL‑2 pathway. Oncology Letters, 31, 219. https://doi.org/10.3892/ol.2026.15574
MLA
Chen, X., Li, H., Cheng, X., Cai, Y., Guo, G."Exosomal‑miR‑32‑5p directly targets FOXN2 to regulate the proliferation, migration and apoptosis of uterine corpus endometrial carcinoma via the PI3K/AKT/BCL‑2 pathway". Oncology Letters 31.6 (2026): 219.
Chicago
Chen, X., Li, H., Cheng, X., Cai, Y., Guo, G."Exosomal‑miR‑32‑5p directly targets FOXN2 to regulate the proliferation, migration and apoptosis of uterine corpus endometrial carcinoma via the PI3K/AKT/BCL‑2 pathway". Oncology Letters 31, no. 6 (2026): 219. https://doi.org/10.3892/ol.2026.15574
Copy and paste a formatted citation
x
Spandidos Publications style
Chen X, Li H, Cheng X, Cai Y and Guo G: Exosomal‑miR‑32‑5p directly targets FOXN2 to regulate the proliferation, migration and apoptosis of uterine corpus endometrial carcinoma via the PI3K/AKT/BCL‑2 pathway. Oncol Lett 31: 219, 2026.
APA
Chen, X., Li, H., Cheng, X., Cai, Y., & Guo, G. (2026). Exosomal‑miR‑32‑5p directly targets FOXN2 to regulate the proliferation, migration and apoptosis of uterine corpus endometrial carcinoma via the PI3K/AKT/BCL‑2 pathway. Oncology Letters, 31, 219. https://doi.org/10.3892/ol.2026.15574
MLA
Chen, X., Li, H., Cheng, X., Cai, Y., Guo, G."Exosomal‑miR‑32‑5p directly targets FOXN2 to regulate the proliferation, migration and apoptosis of uterine corpus endometrial carcinoma via the PI3K/AKT/BCL‑2 pathway". Oncology Letters 31.6 (2026): 219.
Chicago
Chen, X., Li, H., Cheng, X., Cai, Y., Guo, G."Exosomal‑miR‑32‑5p directly targets FOXN2 to regulate the proliferation, migration and apoptosis of uterine corpus endometrial carcinoma via the PI3K/AKT/BCL‑2 pathway". Oncology Letters 31, no. 6 (2026): 219. https://doi.org/10.3892/ol.2026.15574
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