miR-148a-3p Targeting SMURF2 in Regulating Osteogenic Differentiation and Enamel Development during In Vitro Tooth Organogenesis

Qian XU; Yumei CUI; Siming MA; Yunhong LIN; Yijing XIONG; Zijun SONG; Xudong LI

doi:10.12259/j.issn.2095-610X.S20231103

Volume 44 Issue 11

Nov. 2023

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Qian XU, Yumei CUI, Siming MA, Yunhong LIN, Yijing XIONG, Zijun SONG, Xudong LI. miR-148a-3p Targeting SMURF2 in Regulating Osteogenic Differentiation and Enamel Development during In Vitro Tooth Organogenesis[J]. Journal of Kunming Medical University, 2023, 44(11): 16-21. doi: 10.12259/j.issn.2095-610X.S20231103

Citation:

Qian XU, Yumei CUI, Siming MA, Yunhong LIN, Yijing XIONG, Zijun SONG, Xudong LI. miR-148a-3p Targeting SMURF2 in Regulating Osteogenic Differentiation and Enamel Development during In Vitro Tooth Organogenesis[J]. Journal of Kunming Medical University, 2023, 44(11): 16-21. doi: 10.12259/j.issn.2095-610X.S20231103

Citation:

Qian XU, Yumei CUI, Siming MA, Yunhong LIN, Yijing XIONG, Zijun SONG, Xudong LI. miR-148a-3p Targeting SMURF2 in Regulating Osteogenic Differentiation and Enamel Development during In Vitro Tooth Organogenesis[J]. Journal of Kunming Medical University, 2023, 44(11): 16-21. doi: 10.12259/j.issn.2095-610X.S20231103

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miR-148a-3p Targeting SMURF2 in Regulating Osteogenic Differentiation and Enamel Development during In Vitro Tooth Organogenesis

doi: 10.12259/j.issn.2095-610X.S20231103

Dept. of Oral Restorations，Affiliated Stomatological Hospital of Kunming Medical University，Kunming Yunnan 650106，China

Received Date: 2023-09-06
Available Online: 2023-11-13
Publish Date: 2023-11-30

Abstract

Abstract

Objective To explore the role of miR-148a-3p in osteogenic differentiation and enamel development during in vitro tooth organogenesis and uncover its underlying molecular mechanism. Methods Human dental pulp stem cells and oral epithelial cells were obtained. Human dental pulp stem cells were transfected with miR-148a-3p mimics, miR-148a-3p inhibitors, or a negative control. A three-dimensional co-culture system was established by seeding human dental pulp stem cells onto the Matrigel matrix and overlaying them with oral epithelial cells. The impact of miR-148a-3p on cell proliferation and protein expression was assessed using MTT assay. The expression levels of osteogenic marker RUNX2 and enamel development marker E-cadherin were determined through Western blotting. The interaction between SMURF2（SMAD-specific E3 ubiquitin-protein ligase 2） and miR-148a-3p was validated via a luciferase reporter assay. Results Downregulation of miR-148a-3p using its inhibitor in human dental pulp stem cells led to reduced cell viability in the 3D co-culture system（P < 0.05）. Conversely, upregulation of miR-148a-3p using its mimic increased the expression of osteogenic marker RUNX2 and enamel development marker E-cadherin in the co-culture setting（P < 0.05）. TargetScan software predicted a binding site for miR-148a-3p within the 3’ -UTR of SMURF2. Luciferase report Experiments showed that miR-148a-3p mimics significantly inhibited the luciferase activity of wild-type（P < 0.05）, while Western blot results showed that miR-148a-3p mimics significantly down-regulated the expression of SMURF2（P < 0.05）. Conclusion The research findings suggest that miR-148a-3p potentially regulates the expression of RUNX2 and E-cadherin by targeting SMURF2, thereby participating in osteogenic differentiation of human dental pulp stem cells and enamel development in oral epithelial cells during in vitro tooth organogenesis. This study provides promising therapeutic targets for dental repair and treatment.
- MiR-148a-3p,
- Dental pulp stem cells,
- Osteogenic differentiation,
- Enamel development,
- SMURF2

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