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Hsa_circ_0122913 suppresses the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells and is involved in the effects of icariin treatment via sponging miR‑501‑5p

  • Authors:
    • Yeqing Wang
    • Minbo Liu
    • Changhua Liu
    • Huafeng Hong
  • View Affiliations / Copyright

    Affiliations: Department of Pharmacy, Xiaoshan Affiliated Hospital of Wenzhou Medical University, Hangzhou, Zhejiang 311200, P.R. China, Department of Osteoporosis Care and Control, Xiaoshan Affiliated Hospital of Wenzhou Medical University, Hangzhou, Zhejiang 311200, P.R. China
    Copyright: © Wang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 177
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    Published online on: September 16, 2025
       https://doi.org/10.3892/br.2025.2055
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Abstract

Circular RNAs (circRNAs) play significant roles in several biological events. It has been revealed that hsa_circ_0122913 was upregulated in peripheral blood samples from patients with senile osteoporotic vertebral compression fracture. Therefore, the present study aimed to investigate the role of hsa_circ_0122913 in osteoporosis and more particularly its effect on the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs). In addition, the involvement of hsa_circ_0122913 in the effectiveness of icariin treatment in BMSCs was also explored. For functional experiments, BMSCs were transfected with short hairpin RNAs targeting hsa_circ_0122913 and defective in cullin neddylation 1 domain containing 1 (DCUN1D1). To determine BMSCs viability and apoptosis, Cell Counting Kit 8 (CCK‑8) and Annexin V‑APC/7‑AAD assays were carried out, respectively. Furthermore, the expression levels of hsa_circ_0122913, osteopontin (OPN), runt‑related transcription factor 2 (RUNX2), osterix (OSX) and DCUN1D1 were measured by reverse transcription‑quantitative PCR (RT‑qPCR) and western blot analysis. The CircBank database was used to predict the potential binding sites of microRNA (miR)‑501‑5p οn hsa_circ_0122913. The binding capacity of miR‑501‑5p on hsa_circ_0122913 was then verified by dual‑luciferase assay. Finally, alizarin red staining was used to observe the degree of osteogenic differentiation. The CCK‑8 and Annexin V‑APC/7‑AAD assay results demonstrated that hsa_circ_0122913 knockdown in BMSCs improved cell viability and attenuated cell apoptosis. Additionally, the RT‑qPCR and western blot analysis results revealed that hsa_circ_0122913 silencing could enhance the mRNA and protein expression levels of OSX, RUNX2 and OPN. Furthermore, the results indicated that hsa_circ_0122913 could sponge miR‑501‑5p, while the hsa_circ_0122913/DCUN1D1 axis could be involved in the beneficial effects of icariin on BMSCs and more particularly on promoting cell viability and osteogenic differentiation. Overall, the current study suggested that hsa_circ_0122913 could be a novel biomarker for identifying individuals at high risk of osteoporosis, thus being involved in the prevention and treatment of the disease.
View Figures

Figure 1

Hsa_circ_0122913 suppresses the
proliferation and osteogenic differentiation of BMSCs in
vitro. (A) The expression of knocked down hsa_circ_0122913 in
BMSCs determined by RT-qPCR. (B) The cell viability of
hsa_circ_0122913 knocking-down in BMSCs after 48 h, 72 h and 7 d
determined by Cell Counting Kit-8 assay. (C) The apoptotic rate of
hsa_circ_0122913 knocking-down in BMSCs group after 48, 72 h and 7
d determined by flow cytometric analysis. (D) The osteogenic
differentiation ability after 7 days of osteogenic induction
detected by alizarin staining. (E) The mRNA expression of OSX,
RUNX2 and OPN in BMSCs after knockdown of hsa_circ_0122913 detected
by RT-qPCR. (F) The protein expression of OSX, RUNX2 and OPN in
BMSCs after knockdown of hsa_circ_0122913 detected by western blot
analysis. *P<0.05, **P<0.01 and
***P<0.001 vs. NC. BMSCs, bone marrow mesenchymal
stem cells; RT-qPCR, reverse transcription-quantitative PCR; OSX,
osterix; RUNX2, runt-related transcription factor 2; OPN,
osteopontin; NC, negative control.

Figure 2

Hsa_circ_0122913 is a competing
endogenous RNA for miR-501-5p. (A) The binding sites between
hsa_circ_0122913 and miR-501-5p analyzed by circBANK database. (B)
The target relationship of hsa_circ_0122913 and miR-501-5p
indicated by dual-luciferase reporter assay. **P<0.01
vs. NC. miR, microRNA; NC, negative control; NS, not significant;
WT, wild-type; MUT, mutant.

Figure 3

Icariin treatment and DCUN1D1
knockdown could also promote the proliferation and osteogenic
differentiation of BMSCs in vitro. (A) The expression of
DCUN1D1 knocked down in BMSCs determined by reverse
transcription-quantitative PCR. (B) The cell viability of
hsa_circ_0122913 and DCUN1D1 knocked down in BMSCs and icariin
treatment at 24 h, 48 h and 7 days determined by Cell Counting
Kit-8 assay. (C) The osteogenic differentiation ability after three
weeks of osteogenic induction detected by alizarin staining.
*P<0.05, **P<0.01 and
****P<0.0001 vs. NC; #P<0.05 and
####P<0.0001 vs. mock. DCUN1D1, defective in cullin
neddylation 1 domain containing 1; BMSCs, bone marrow mesenchymal
stem cells; NC, negative control; sh-, short hairpin.

Figure 4

Hsa_circ_0122913/DCUN1D1 axis might
participate in the effect of icariin treatment on bone marrow
mesenchymal stem cells. (A) The mRNA level of hsa_circ_0122913
after knockdown of hsa_circ_0122913 and DCUN1D1 and treated with
icariin. (B) The mRNA level of DCUN1D1 after knockdown of
hsa_circ_0122913 and DCUN1D1 and treated with icariin. (C) The
protein level of DCUN1D1 after knockdown of hsa_circ_0122913 and
DCUN1D1 and treated with icariin. ***P<0.001 and
****P<0.0001 vs. NC; ##P<0.01,
###P<0.001 and ####P<0.0001 vs. mock.
DCUN1D1, defective in cullin neddylation 1 domain containing 1; NC,
negative control; sh-, short hairpin.
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Copy and paste a formatted citation
Spandidos Publications style
Wang Y, Liu M, Liu C and Hong H: Hsa_circ_0122913 suppresses the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells and is involved in the effects of icariin treatment via sponging miR‑501‑5p. Biomed Rep 23: 177, 2025.
APA
Wang, Y., Liu, M., Liu, C., & Hong, H. (2025). Hsa_circ_0122913 suppresses the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells and is involved in the effects of icariin treatment via sponging miR‑501‑5p. Biomedical Reports, 23, 177. https://doi.org/10.3892/br.2025.2055
MLA
Wang, Y., Liu, M., Liu, C., Hong, H."Hsa_circ_0122913 suppresses the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells and is involved in the effects of icariin treatment via sponging miR‑501‑5p". Biomedical Reports 23.5 (2025): 177.
Chicago
Wang, Y., Liu, M., Liu, C., Hong, H."Hsa_circ_0122913 suppresses the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells and is involved in the effects of icariin treatment via sponging miR‑501‑5p". Biomedical Reports 23, no. 5 (2025): 177. https://doi.org/10.3892/br.2025.2055
Copy and paste a formatted citation
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Spandidos Publications style
Wang Y, Liu M, Liu C and Hong H: Hsa_circ_0122913 suppresses the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells and is involved in the effects of icariin treatment via sponging miR‑501‑5p. Biomed Rep 23: 177, 2025.
APA
Wang, Y., Liu, M., Liu, C., & Hong, H. (2025). Hsa_circ_0122913 suppresses the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells and is involved in the effects of icariin treatment via sponging miR‑501‑5p. Biomedical Reports, 23, 177. https://doi.org/10.3892/br.2025.2055
MLA
Wang, Y., Liu, M., Liu, C., Hong, H."Hsa_circ_0122913 suppresses the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells and is involved in the effects of icariin treatment via sponging miR‑501‑5p". Biomedical Reports 23.5 (2025): 177.
Chicago
Wang, Y., Liu, M., Liu, C., Hong, H."Hsa_circ_0122913 suppresses the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells and is involved in the effects of icariin treatment via sponging miR‑501‑5p". Biomedical Reports 23, no. 5 (2025): 177. https://doi.org/10.3892/br.2025.2055
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