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Effect of the miR‑26a/b‑5p‑STAT3‑YKL‑40 regulatory axis on proliferation, migration, and invasion of endometrial cancer cells

  • Authors:
    • Xinzhao Sun
    • Shanshan Lin
    • Xueyan Zhong
    • Yanlu Luo
    • Jiahuang Yang
    • Jiangtao Fan
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    Affiliations: Department of Gynecology, The First Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi 530021, P.R. China
    Copyright: © Sun et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 127
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    Published online on: May 8, 2026
       https://doi.org/10.3892/or.2026.9132
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Abstract

The incidence of endometrial cancer (EC) is on the rise annually, emphasizing the importance of timely diagnosis and treatment. Signal transducer and activator of transcription 3 (STAT3) has been identified as a proto‑oncogene involved in multiple signaling pathways affecting various biological processes, especially in tumours. The aim of the present study was to validate the impact of STAT3 on EC phenotype and investigate its upstream and downstream regulatory mechanisms. Immunohistochemical analysis was conducted to assess STAT3 expression in EC tissues, followed by dual luciferase assays to confirm the regulatory relationship between STAT3 and microRNA (miR)‑26a/b‑5p. The effects of STAT3 and miR‑26a/b‑5p on HEC‑1A cell proliferation and metastasis were evaluated through Cell Counting Kit‑8 assays, cell scratch assays, and Transwell assays. Co‑immunoprecipitation assays verified the binding between STAT3 and chitinase‑3‑like protein 1 (CHI3L1, also known as YKL‑40). The results demonstrated high STAT3 expression in EC, associated with disease progression. miR‑26a/b‑5p directly targeted STAT3. Upregulated STAT3 enhanced HEC‑1A cell proliferation, migration, and invasion, counteracting the inhibitory effects of miR‑26a/b‑5p on cell migration and invasion. In addition, it was confirmed that STAT3 binds to and promotes the expression of YKL‑40. These findings contribute to clarifying the pathogenesis of EC.
View Figures

Figure 1

Technical roadmap of the study. miR,
microRNA; RT-qPCR, reverse transcription-quantitative PCR; STAT3,
signal transducer and activator of transcription 3; WB, western
blotting; OE, overexpression; EC, endometrial cancer; IHC,
immunohistochemistry; Co-IP, co-immunoprecipitation; YKL-40,
chitinase-3-like protein 1; KD, knockdown.

Figure 2

Expression of STAT3 in EC tissues.
(A) The STAT3 expression profile in EC tissues (n=45) and
paracancerous endometrial tissues (n=40) was detected using IHC
staining. (B) Quantitative analysis of STAT3 expression in EC
tissues (n=45) and para-carcinoma intima tissues (n=40). STAT3,
signal transducer and activator of transcription 3; EC, endometrial
cancer; AOD, average optical density.

Figure 3

STAT3 promotes proliferation and
migration of HEC-1A cells. (A and B) Western blotting verified the
overexpression and silencing efficacy of STAT3. (C and D) Cell
Counting Kit-8 assay revealed that overexpression of STAT3 promoted
cell proliferation, whereas knockdown of STAT3 had the opposite
effect. (E and F) Overexpression of STAT3 promoted cell wound
healing, whereas knockdown of STAT3 had the opposite effect. (G-J)
Transwell assay showed that overexpression of STAT3 promoted cell
migration and invasion, while knockdown exerted the opposite
effect. *P<0.05, **P<0.01 and ***P<0.001. STAT3, signal
transducer and activator of transcription 3; sh-, short hairpin;
NC, negative control.

Figure 4

miR-26a/b-5p targets STAT3. (A) STAT3
wild-type vector and mutant vector sequences. (B) Dual luciferase
activity assay. (C and D) Overexpression of miR-26a/b-5p reduces
the mRNA and protein levels of STAT3 in cells. **P<0.01 and
***P<0.001. miR, microRNA; STAT3, signal transducer and
activator of transcription 3; WT, wild-type; MUT, mutant; NC,
negative control; Fluc, firefly luciferase; Rluc, Renilla
luciferase.

Figure 5

miR-26a/b-5p reduces the migration
and invasion activity of HEC-1A cells. (A) miR-26a/b-5p
Transduction efficiency was measured, indicating successful
overexpression of miR-26a/b-5p. (B) Cell Counting Kit-8 cell
proliferation assay, (C) wound healing assay, and (D and E)
Transwell migration and invasion assays were used to evaluate the
effects of miR-26a/b-5p on cell proliferation, migration, and
invasion. *P<0.05, **P<0.01 and ***P<0.001. miR, microRNA;
STAT3, signal transducer and activator of transcription 3; NC,
negative control; ns, not significant.

Figure 6

Overxepression of STAT3 counteracts
the suppressive effect of miR-26a/b-5p on the cellular biological
behavior. (A) STAT3 was reintroduced into cells overexpressing
miR-26a/b-5p, and STAT3 protein expression in each group was
detected by western blotting. (B and C) Cell Counting Kit-8 assay,
(D) wound healing assay, and (E and F) Transwell migration and
invasion assays were used to determine whether STAT3 overexpression
reversed the inhibitory effects of miR-26a/b-5p on cell
proliferation, migration, and invasion. *P<0.05, **P<0.01 and
***P<0.001. STAT3, signal transducer and activator of
transcription 3; miR, microRNA; ns, not significant.

Figure 7

STAT3 modulates YKL-40 expression.
(A) Expression analysis of YKL-40 in tumor and non-tumor samples
from the GEPIA2 database. (B) STRING database analysis of the
protein-protein interaction between STAT3 and CHI3L1 (the gene
encoding YKL-40). Purple lines indicate experimentally determined
interactions, green lines represent text-mining evidence, and gray
lines denote co-expression evidence. (C) Three-dimensional
interaction between YKL-40 and STAT3. (D) Co-immunoprecipitation
assay showing the interaction of STAT3 and YKL-40. (E and F) STAT3
overexpression induced YKL-40 levels. (G and H) Overexpression of
miR-26a/b-5p led to downregulation of YKL-40 mRNA and protein
expression levels. *P<0.05 and
**P<0.01. STAT3, signal transducer and activator of
transcription 3; CHI3L1 and YKL-40, chitinase-3-like protein 1; IP,
immunoprecipitation; NC, negative control; miR, microRNA.

Figure 8

Pro-carcinogenic effect of STAT3 is
dependent on YKL-40. (A and B) The expression levels of YKL-40 and
STAT3 in each group of cells was quantified using western blotting.
Results of the (C) Cell Counting Kit-8 assay, (D) cell scratch
assay, and (E and F) Transwell migration and invasion assays.
*P<0.05, **P<0.01 and ***P<0.001. STAT3, signal transducer
and activator of transcription 3; YKL-40, chitinase-3-like protein
1; sh, short hairpin; NC, negative control; ns, not
significant.

Figure 9

Schematic diagram of the proposed
mechanism of STAT3 in endometrial cancer. miR, microRNA; STAT3,
signal transducer and activator of transcription 3; EC, endometrial
cancer; YKL-40, chitinase-3-like protein 1; OE, overexpression.
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Copy and paste a formatted citation
Spandidos Publications style
Sun X, Lin S, Zhong X, Luo Y, Yang J and Fan J: Effect of the miR‑26a/b‑5p‑STAT3‑YKL‑40 regulatory axis on proliferation, migration, and invasion of endometrial cancer cells. Oncol Rep 56: 127, 2026.
APA
Sun, X., Lin, S., Zhong, X., Luo, Y., Yang, J., & Fan, J. (2026). Effect of the miR‑26a/b‑5p‑STAT3‑YKL‑40 regulatory axis on proliferation, migration, and invasion of endometrial cancer cells. Oncology Reports, 56, 127. https://doi.org/10.3892/or.2026.9132
MLA
Sun, X., Lin, S., Zhong, X., Luo, Y., Yang, J., Fan, J."Effect of the miR‑26a/b‑5p‑STAT3‑YKL‑40 regulatory axis on proliferation, migration, and invasion of endometrial cancer cells". Oncology Reports 56.1 (2026): 127.
Chicago
Sun, X., Lin, S., Zhong, X., Luo, Y., Yang, J., Fan, J."Effect of the miR‑26a/b‑5p‑STAT3‑YKL‑40 regulatory axis on proliferation, migration, and invasion of endometrial cancer cells". Oncology Reports 56, no. 1 (2026): 127. https://doi.org/10.3892/or.2026.9132
Copy and paste a formatted citation
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Spandidos Publications style
Sun X, Lin S, Zhong X, Luo Y, Yang J and Fan J: Effect of the miR‑26a/b‑5p‑STAT3‑YKL‑40 regulatory axis on proliferation, migration, and invasion of endometrial cancer cells. Oncol Rep 56: 127, 2026.
APA
Sun, X., Lin, S., Zhong, X., Luo, Y., Yang, J., & Fan, J. (2026). Effect of the miR‑26a/b‑5p‑STAT3‑YKL‑40 regulatory axis on proliferation, migration, and invasion of endometrial cancer cells. Oncology Reports, 56, 127. https://doi.org/10.3892/or.2026.9132
MLA
Sun, X., Lin, S., Zhong, X., Luo, Y., Yang, J., Fan, J."Effect of the miR‑26a/b‑5p‑STAT3‑YKL‑40 regulatory axis on proliferation, migration, and invasion of endometrial cancer cells". Oncology Reports 56.1 (2026): 127.
Chicago
Sun, X., Lin, S., Zhong, X., Luo, Y., Yang, J., Fan, J."Effect of the miR‑26a/b‑5p‑STAT3‑YKL‑40 regulatory axis on proliferation, migration, and invasion of endometrial cancer cells". Oncology Reports 56, no. 1 (2026): 127. https://doi.org/10.3892/or.2026.9132
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