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Enhanced SLC2A4 expression mitigates insulin resistance in gestational diabetes mellitus via the FoxO signaling pathway in vitro

  • Authors:
    • Yanqi Jin
    • Xiaoyan Cheng
    • Jinting Wu
    • Yi Zhu
    • Hui Xu
    • Chunyu Cao
  • View Affiliations / Copyright

    Affiliations: Department of Obstetrics and Gynecology, Affiliated Maternity and Child Health Care Hospital of Nantong University, Nantong, Jiangsu 226007, P.R. China, Nantong Institute of Genetics and Reproductive Medicine, Affiliated Maternity and Child Health Care Hospital of Nantong University, Nantong, Jiangsu 226007, P.R. China
    Copyright: © Jin et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 206
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    Published online on: August 22, 2025
       https://doi.org/10.3892/etm.2025.12956
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Abstract

Gestational diabetes mellitus (GDM), a type of diabetes mellitus occurring in pregnant women, increases the risk of birth trauma. Solute carrier family 2 member 4 (SLC2A4) polymorphism is notably associated with GDM susceptibility; however, the mechanism is unknown. In the present study, HTR‑8/SVneo cells were treated with high glucose concentrations and transfected with SLC2A4 and Forkhead box O (FoxO)1 to investigate their roles in the insulin (INS) resistance of GDM trophoblast cells. The results showed that the expression levels of SLC2A4, INS receptor (INSR) and INS were significantly decreased in HTR‑8/SVneo cells under hyperglycemic conditions. Furthermore, overexpression of SLC2A4 reduced the levels of phosphorylated FoxO (p‑FoxO)1, FoxO1, p‑FoxO3a and FoxO3a but enhanced the expression levels of INS and INSR, improving glucose uptake within the hyperglycemic environment. Additionally, both the analysis of the signaling pathways related to SLC2A4 and GDM using the Kyoto Encyclopedia of Genes and Genomes pathway analysis and the results of CCK‑8 determination of cell vitality both emphasized that the FoxO signaling was a pivotal pathway affected by SLC2A4 in GDM. In conclusion, the present study demonstrated that overexpression of SLC2A4 enhanced the INS signaling pathway, as evidenced by western blotting showing increased levels of INSR and INS, along with elevated glucose uptake. Concurrently, SLC2A4 overexpression inhibited the FoxO signaling pathway, as indicated by reduced protein levels of p‑FoxO1, total FoxO1, p‑FoxO3a and total FoxO3a. Collectively, these molecular alterations contribute to the alleviation of insulin resistance in patients with GDM.
View Figures

Figure 1

Level of SLC2A4 in HG-induced
HTR-8/SVneo cells. (A) A screen of targeted genes related to
gestational diabetes mellitus in the CTD, DisGeNet and Genecards
databases. (B) The level of LDH was analyzed using an LDH assay.
(C) The expression of SLC2A4 was analyzed by reverse
transcription-quantitative PCR. (D) The expression of SLC2A4 was
analyzed by western blotting. **P<0.01 and
***P<0.001 vs. control. CTD, Comparative
Toxicogenomics Database; SLC2A4, solute carrier family 2 member 4;
LDH, lactate dehydrogenase; HG, high glucose.

Figure 2

Effect of SLC2A4 overexpression on
HG-induced HTR-8/SVneo cells. The expression of SLC2A4 was analyzed
by (A) RT-qPCR and (B) western blotting. The expression of INS and
INSR was analyzed by (C) RT-qPCR and (D) western blotting. (E)
Glucose uptake was detected using a glucose uptake assay. (F) Cell
viability was measured using a Cell Counting Kit-8 assay.
***P<0.001 vs. Control; nsP>0.05 vs.
HG; #P<0.05 and ###P<0.001 vs. HG +
pc-NC. SLC2A4, solute carrier family 2 member 4; HG, high glucose;
RT-qPCR, reverse transcription-quantitative PCR; ns, not
significant; NC, negative control; OD, optical density; INS,
insulin; INSR, INS receptor.

Figure 3

Role of the FoxO signaling pathway in
pc-SLC2A4-induced changes in HG-induced HTR-8/SVneo cells. (A)
Screen of targeted pathways related to gestational diabetes
mellitus and SLC2A4 (five common genes identified through the
CTD/GeneCards/DisGeNet database) using Kyoto Encyclopedia of Genes
and Genomes, suggesting a critical role for FoxO signaling pathway.
(B) The expression levels of p-FoxO1, FoxO1, p-FoxO3a and FoxO3a
were analyzed by western blotting, GAPDH was used as the internal
control for the results. The expression of INS and INSR was
analyzed by (C) reverse transcription-quantitative PCR and (D)
western blotting. (E) Glucose uptake was detected by glucose uptake
assay. (F) Cell viability was measured using a Cell Counting Kit-8
assay. ***P<0.001 vs. Control; nsP>0.05
vs. HG; ###P<0.001 vs. HG + pc-NC;
$P<0.05, $$P<0.01 and
$$$P<0.001 vs. HG + pc-SLC2A4. SLC2A4, solute carrier
family 2 member 4; HG, high glucose; ns, not significant; NC,
negative control; OD, optical density; INS, insulin; INSR, INS
receptor; p, phosphorylated; FoxO, Forkhead box O.
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Spandidos Publications style
Jin Y, Cheng X, Wu J, Zhu Y, Xu H and Cao C: Enhanced SLC2A4 expression mitigates insulin resistance in gestational diabetes mellitus via the FoxO signaling pathway <em>in vitro</em>. Exp Ther Med 30: 206, 2025.
APA
Jin, Y., Cheng, X., Wu, J., Zhu, Y., Xu, H., & Cao, C. (2025). Enhanced SLC2A4 expression mitigates insulin resistance in gestational diabetes mellitus via the FoxO signaling pathway <em>in vitro</em>. Experimental and Therapeutic Medicine, 30, 206. https://doi.org/10.3892/etm.2025.12956
MLA
Jin, Y., Cheng, X., Wu, J., Zhu, Y., Xu, H., Cao, C."Enhanced SLC2A4 expression mitigates insulin resistance in gestational diabetes mellitus via the FoxO signaling pathway <em>in vitro</em>". Experimental and Therapeutic Medicine 30.5 (2025): 206.
Chicago
Jin, Y., Cheng, X., Wu, J., Zhu, Y., Xu, H., Cao, C."Enhanced SLC2A4 expression mitigates insulin resistance in gestational diabetes mellitus via the FoxO signaling pathway <em>in vitro</em>". Experimental and Therapeutic Medicine 30, no. 5 (2025): 206. https://doi.org/10.3892/etm.2025.12956
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Spandidos Publications style
Jin Y, Cheng X, Wu J, Zhu Y, Xu H and Cao C: Enhanced SLC2A4 expression mitigates insulin resistance in gestational diabetes mellitus via the FoxO signaling pathway <em>in vitro</em>. Exp Ther Med 30: 206, 2025.
APA
Jin, Y., Cheng, X., Wu, J., Zhu, Y., Xu, H., & Cao, C. (2025). Enhanced SLC2A4 expression mitigates insulin resistance in gestational diabetes mellitus via the FoxO signaling pathway <em>in vitro</em>. Experimental and Therapeutic Medicine, 30, 206. https://doi.org/10.3892/etm.2025.12956
MLA
Jin, Y., Cheng, X., Wu, J., Zhu, Y., Xu, H., Cao, C."Enhanced SLC2A4 expression mitigates insulin resistance in gestational diabetes mellitus via the FoxO signaling pathway <em>in vitro</em>". Experimental and Therapeutic Medicine 30.5 (2025): 206.
Chicago
Jin, Y., Cheng, X., Wu, J., Zhu, Y., Xu, H., Cao, C."Enhanced SLC2A4 expression mitigates insulin resistance in gestational diabetes mellitus via the FoxO signaling pathway <em>in vitro</em>". Experimental and Therapeutic Medicine 30, no. 5 (2025): 206. https://doi.org/10.3892/etm.2025.12956
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