MiR-346 Inhibits Chondrocyte Proliferation in Idiopathic Short Stature by Targeting SMAD3

Jing REN; Yani ZENG; Zhirong SHANG

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Jing REN, Yani ZENG, Zhirong SHANG. MiR-346 Inhibits Chondrocyte Proliferation in Idiopathic Short Stature by Targeting SMAD3[J]. Journal of Kunming Medical University.

Citation:

Jing REN, Yani ZENG, Zhirong SHANG. MiR-346 Inhibits Chondrocyte Proliferation in Idiopathic Short Stature by Targeting SMAD3[J]. Journal of Kunming Medical University.

Jing REN, Yani ZENG, Zhirong SHANG. MiR-346 Inhibits Chondrocyte Proliferation in Idiopathic Short Stature by Targeting SMAD3[J]. Journal of Kunming Medical University.

Citation:

Jing REN, Yani ZENG, Zhirong SHANG. MiR-346 Inhibits Chondrocyte Proliferation in Idiopathic Short Stature by Targeting SMAD3[J]. Journal of Kunming Medical University.

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MiR-346 Inhibits Chondrocyte Proliferation in Idiopathic Short Stature by Targeting SMAD3

1.
Department of Child Health Care
2.
Department of Pregnancy and Child Health Care
3.
Department of Laboratory Medicine，Mianyang Maternal and Child Health Hospital/Mianyang Children’s Hospital，Mianyang Sichuan 621000，China

Received Date: 2026-03-17
Available Online: 2026-10-28

Abstract

Abstract

Objective To investigate the expression of miR-346 in patients with idiopathic short stature （ISS） and its effect on chondrocyte proliferation, and to elucidate the molecular mechanism by which it acts through targeted regulation of SMAD family member 3（SMAD3）. Methods A case-control study design was adopted. Patients with ISS admitted to Mianyang Children's Hospital between December 2019 and December 2023 were enrolled as the ISS group （n = 38）. Meanwhile, children whose height was within ±1 standard deviation of the average level of the population of the same age, gender and ethnicity were recruited as the control group （n = 38）, and their peripheral blood samples were collected. The serum expression levels of miR-346 and SMAD3 were detected by quantitative real-time polymerase chain reaction （qRT-PCR）. Human chondrocytes C28/I2 were cultured in vitro, and four experimental groups were established: miR-NC group （transfected with negative control miR-NC）, miR-346 inhibitor group （transfected with miR-346 inhibitor）, miR-346 inhibitor + si-NC group （co-transfected with miR-346 inhibitor and negative control si-NC）, and miR-346 inhibitor + si-SMAD3 group （co-transfected with miR-346 inhibitor and si-SMAD3）. Changes in Cell proliferation capacity were assessed using the CCK-8 assay and Edu staining. Cell cycle distribution was analyzed by flow cytometry. Protein expression levels of cell cycle phases and osteogenic differentiation-related proteins, as well as SMAD3, were measured by western blotting. The targeting relationship between miR-346 and SMAD3 was predicted using bioinformatics and validated through dual-luciferase reporter gene assays. Results Compared with the control group, the serum expression level of miR - 346 in the ISS group was significantly increased, while the expression level of SMAD3 was significantly decreased （P < 0.05）. Compared with the miR-NC group, the miR-346 expression levels, the proportions of cells in the G₁ and S phases, and the p21 protein expression levels in cells of the miR-346 inhibitor group and the miR-346 inhibitor + si-NC group were significantly decreased （P < 0.05）. In contrast, cell proliferation viability, the proportion of EDU-positive cells, the proportion of cells in the G₂ phase, and the protein expression levels of Cyclin D1, CDK4, CDK6, MMP-13, COL10A, RUNX2, OCN, and OPN, as well as the mRNA expression of SMAD3 and the p-SMAD3 protein level, were significantly increased （P < 0.05）. Compared with the miR-346 inhibitor group and the miR-346 inhibitor + si-NC group, the proportions of cells in the G₁ and S phases and the p21 protein expression level in cells of the miR-346 inhibitor + si-SMAD3 group were significantly increased （P < 0.05）. Conversely, cell proliferation viability, EDU-positive ratio, the proportion of cells in the G2 phase, and the protein expression levels of Cyclin D1, CDK4, CDK6, MMP-13, COL10A, RUNX2, OCN, and OPN, as well as the mRNA expression of SMAD3 and the p-SMAD3 protein level, were significantly decreased （P < 0.05）. Conclusion miR-346 expression is up-regulated in ISS patients and may interfere with chondrocyte proliferation and osteogenic differentiation by targeted inhibition of SMAD3.
- miR-346,
- SMAD3,
- Idiopathic short stature,
- Chondrocyte,
- Proliferation,
- Osteogenesis

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References(25)

References

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