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Article Open Access

Circular RNA hsa_circ_0000118 promotes atrial fibrosis by regulating the microRNA‑34a‑5p/Smad4 axis

  • Authors:
    • Na Wu
    • Yuanbo Zhang
    • Yuhong Zeng
    • Lanqing Yang
    • Jun Li
    • Yanxiu Chen
    • Zhiquan Yuan
    • Xinghua Chen
    • Chengying Li
    • Long Wu
    • Tongjian Cai
    • Zhihui Zhang
    • Li Zhong
    • Yafei Li
  • View Affiliations / Copyright

    Affiliations: Department of Epidemiology, College of Preventive Medicine, Army Medical University (Third Military Medical University), Chongqing 400038, P.R. China, Department of Cardiothoracic Surgery, Southwest Hospital, The First Affiliated Hospital of Army Medical University (Third Military Medical University), Chongqing 400038, P.R. China, Department of Cardiology, Southwest Hospital, The First Affiliated Hospital of Army Medical University (Third Military Medical University), Chongqing 400038, P.R. China, Department of Cardiovascular Medicine, Third Affiliated Hospital of Chongqing Medical University, Chongqing 401120, P.R. China
    Copyright: © Wu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 339
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    Published online on: October 2, 2025
       https://doi.org/10.3892/mmr.2025.13704
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Abstract

The regulatory functions and underlying mechanisms of circular RNAs (circRNAs/circs) in atrial fibrillation (AF) are largely unknown. The present study aimed to assess the prognostic roles and potential biological functions of hsa_circ_0000118 in structural remodeling in AF. Gain‑ and loss‑of‑function cell models were established in mouse cardiac fibroblasts. Cell Counting Kit‑8 and Transwell assays were used to analyze the proliferation and migration of cardiac fibroblasts. To evaluate the underlying mechanisms, RNA immunoprecipitation and luciferase reporter assays were performed. hsa_circ_0000118 was demonstrated to be significantly upregulated in atrial tissues of patients with valvular heart disease with AF compared to those without AF. Elevated plasma levels of hsa_circ_0000118 were independently associated with poor prognosis in patients with AF. In vitro, hsa_circ_0000118 promoted collagen I and collagen III expression, and the migration of cardiac fibroblasts. Functional assays demonstrated that hsa_circ_0000118 acted as a competing endogenous RNA of microRNA (miR)‑34a‑5p to reduce the suppressive effect of miR‑34a‑5p on its target Smad4 in cardiac fibroblasts. In conclusion, hsa_circ_0000118 may serve as a non‑invasive prognostic biomarker for AF. The newly identified roles of hsa_circ_0000118 in AF provide a mechanistic understanding of structural remodeling in AF and pave the way towards novel therapies.
View Figures

Figure 1

Expression of hsa_circ_0000118 in AF.
(A) Expression levels of hsa_circ_0000118 in 34 pairs of age- and
sex-matched atrial tissues from patients with and without AF
(unpaired t-test). (B) hsa_circ_0000118 and MAN1A2 expression with
and without RNase R treatment (unpaired t-test). (C)
hsa_circ_0000118 expression in different types of AF (paroxysmal
AF, n=60; persistent AF, n=120; and permanent AF, n=12; one-way
ANOVA followed by the Sidak method). (D) Kaplan-Meier event-free
survival curves for relative hsa_circ_0000118 expression in plasma.
*P<0.05. AF, atrial fibrillation; circ, circular RNA; MAN1A2,
mannosidase α class 1A member 2.

Figure 2

Effect of mmu_circ_0010297
overexpression and knockdown on the proliferation and migration of
mouse cardiac fibroblasts. (A) Overexpression and knockdown
efficiency of mmu_circ_0010297 in cardiac fibroblasts [unpaired
t-test (left) and one-way ANOVA followed by the Sidak method
(right)]. (B) Growth curves of cardiac fibroblasts after
transfection with mmu_circ_0010297 or EV (two-way ANOVA). (C)
Migration of cardiac fibroblasts after transfection with
mmu_circ_0010297 or EV (unpaired t-test). (D) Growth curves of
cardiac fibroblasts after transfection with mmu_circ_0010297 siRNAs
or siNC (two-way ANOVA). (E) Migration of cardiac fibroblasts after
transfection with mmu_circ_0010297 siRNAs or siNC (magnification,
×100×; one-way ANOVA followed by the Sidak method). *P<0.05,
***P<0.001, ****P<0.0001 vs. EV or siNC). circ, circular RNA;
EV, empty vector; NC, negative control; OD, optical density; OE,
overexpression; siRNA/si, small interfering RNA.

Figure 3

Effect of mmu_circ_0010297
overexpression and knockdown on differentiation into myofibroblasts
and collagen expression. (A) Relative Acta2, Vim, Col1a1 and Col3a1
mRNA expression of cardiac fibroblasts after transfection with
mmu_circ_0010297 or EV (unpaired t-test). (B) Relative Acta2, Vim,
Col1a1 and Col3a1 mRNA expression of cardiac fibroblasts after
transfection with mmu_circ_0010297 siRNAs or siNC (one-way ANOVA
followed by the Sidak method). (C) Relative Col1a1 and Col3a1
protein expression of cardiac fibroblasts after transfection with
mmu_circ_0010297 OE or EV (unpaired t-test). (D) Relative Col1a1
and Col3a1 protein expression of cardiac fibroblasts after
transfection with mmu_circ_0010297 siRNAs or siNC (one-way ANOVA
followed by the Sidak method). *P<0.05, **P<0.01,
***P<0.001, ****P<0.0001 vs. EV or siNC. circ, circular RNA;
Col1a1, collagen type I α1 chain; Col3a1, collagen type III α1
chain; EV, empty vector; NC, negative control; OE, overexpression;
siRNA/si, small interfering RNA; Vim, Vimentin.

Figure 4

Examination of the specific mechanism
of mmu_circ_0010297 in regulating collagen production. Expression
of the candidate miRNAs after (A) overexpression of
mmu_circ_0010297 (unpaired t-test) or (B) knockdown of
mmu_circ_0010297 (one-way ANOVA and the Sidak method). (C)
mmu_circ_0010297 and miR-34a-5p levels were determined using
reverse transcription-quantitative PCR after Ago2 or IgG RNA
immunoprecipitation assays (unpaired t-test, as only the Ago2 and
IgG groups were compared). (D) Dual-luciferase reporter assay
(unpaired t-test). (E) Regulatory relationship between Smad4 and
mmu_circ_0010297. For the comparison of OE and EV, an unpaired
t-test was used. For the comparison of si3, si5 and siNC, one-way
ANOVA and the Sidak method were used. (F) Relative expression
levels of Smad4 in cardiac fibroblasts were determined by RT-qPCR
after co-transfection with mmu_circ_0010297 or control EV and
miRNA-34a-5p mimics or NC mimics (one-way ANOVA and the Sidak
method). *P<0.05, **P<0.01, ****P<0.0001 vs. EV, siNC or
Anti-IgG. Ago2, argonaute 2; circ, circular RNA; EV, empty vector;
miRNA/miR, microRNA; MUT, mutant; NC, negative control; OE,
overexpression; si, small interfering RNA; WT, wild-type; RT-q,
reverse transcription-quantitative; ns, not significant.
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Copy and paste a formatted citation
Spandidos Publications style
Wu N, Zhang Y, Zeng Y, Yang L, Li J, Chen Y, Yuan Z, Chen X, Li C, Wu L, Wu L, et al: Circular RNA hsa_circ_0000118 promotes atrial fibrosis by regulating the microRNA‑34a‑5p/Smad4 axis. Mol Med Rep 32: 339, 2025.
APA
Wu, N., Zhang, Y., Zeng, Y., Yang, L., Li, J., Chen, Y. ... Li, Y. (2025). Circular RNA hsa_circ_0000118 promotes atrial fibrosis by regulating the microRNA‑34a‑5p/Smad4 axis. Molecular Medicine Reports, 32, 339. https://doi.org/10.3892/mmr.2025.13704
MLA
Wu, N., Zhang, Y., Zeng, Y., Yang, L., Li, J., Chen, Y., Yuan, Z., Chen, X., Li, C., Wu, L., Cai, T., Zhang, Z., Zhong, L., Li, Y."Circular RNA hsa_circ_0000118 promotes atrial fibrosis by regulating the microRNA‑34a‑5p/Smad4 axis". Molecular Medicine Reports 32.6 (2025): 339.
Chicago
Wu, N., Zhang, Y., Zeng, Y., Yang, L., Li, J., Chen, Y., Yuan, Z., Chen, X., Li, C., Wu, L., Cai, T., Zhang, Z., Zhong, L., Li, Y."Circular RNA hsa_circ_0000118 promotes atrial fibrosis by regulating the microRNA‑34a‑5p/Smad4 axis". Molecular Medicine Reports 32, no. 6 (2025): 339. https://doi.org/10.3892/mmr.2025.13704
Copy and paste a formatted citation
x
Spandidos Publications style
Wu N, Zhang Y, Zeng Y, Yang L, Li J, Chen Y, Yuan Z, Chen X, Li C, Wu L, Wu L, et al: Circular RNA hsa_circ_0000118 promotes atrial fibrosis by regulating the microRNA‑34a‑5p/Smad4 axis. Mol Med Rep 32: 339, 2025.
APA
Wu, N., Zhang, Y., Zeng, Y., Yang, L., Li, J., Chen, Y. ... Li, Y. (2025). Circular RNA hsa_circ_0000118 promotes atrial fibrosis by regulating the microRNA‑34a‑5p/Smad4 axis. Molecular Medicine Reports, 32, 339. https://doi.org/10.3892/mmr.2025.13704
MLA
Wu, N., Zhang, Y., Zeng, Y., Yang, L., Li, J., Chen, Y., Yuan, Z., Chen, X., Li, C., Wu, L., Cai, T., Zhang, Z., Zhong, L., Li, Y."Circular RNA hsa_circ_0000118 promotes atrial fibrosis by regulating the microRNA‑34a‑5p/Smad4 axis". Molecular Medicine Reports 32.6 (2025): 339.
Chicago
Wu, N., Zhang, Y., Zeng, Y., Yang, L., Li, J., Chen, Y., Yuan, Z., Chen, X., Li, C., Wu, L., Cai, T., Zhang, Z., Zhong, L., Li, Y."Circular RNA hsa_circ_0000118 promotes atrial fibrosis by regulating the microRNA‑34a‑5p/Smad4 axis". Molecular Medicine Reports 32, no. 6 (2025): 339. https://doi.org/10.3892/mmr.2025.13704
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