[Micro RNA-451 promoting osteogenesis of mesenchymal stem cells by targeting regulatory calcium binding protein 39]
1Laboratory of Biomechanics, Department of Anatomy, the Third Military Medical University, Chongqing, 400038, P.R.China.
Objective:
To investigate the role of micro RNA-451 (miRNA-451) in promoting the osteogenesis of mesenchymal stem cells (MSCs) by targeting regulatory calcium binding protein 39 (CAB39).
Methods:
pMIR-report and pRL-TK vectors were selected to identify the relationship between miRNA-451 and CAB39 by using dual-luciferase reporter assay. pre-miRNA-451 (group A), anti-miRNA-451 (group C), pre-miRNA negative control (group B), and anti-miRNA negative control (group D) were transfected into the C3H10T1/2 cells, respectively. Then, the cells were collected after osteogenic induction for 7 and 14 days. At 7 and 14 days, the real-time fluorescent quantitative PCR and Western blot assays were performed to detect the related osteogenetic biomarkers [Runx2 and alkaline phosphatase (ALP) mRNA] and expressions of CAB39 protein. At 14 days, the extracellular calcium deposition during the osteogenesis of MSCs was tested by Alizarin red staining method.
Results:
CAB39 was the target gene of miRNA-451. At 7 and 14 days after osteogenic induction, the mRNA expressions of Runx2 and ALP in group A were significantly higher than those in group B (P < 0.05), and the expressions in group C was significantly lower than those in group D (P < 0.05). Furthermore, at 14 days after osteogenic induction, the protein expression of CAB39 in group A (0.55 +/- 0.05) was significantly lower than that in group B (1.00 +/- 0.07), and the protein expression in group C (1.21 +/- 0.05) was significantly higher than that in group D (1.00 +/- 0.04), all showing significant difference (P < 0.05). Finally, at 14 days after osteogenic induction, the extracellular calcium deposition in group A was obviously more than that in group B, and group C was downregulated when compared with group D.
Conclusion:
miRNA-451 can promote the osteogenesis process of MSCs by downregulating the CAB39.
Insights
MicroRNA-451 promotes mesenchymal stem cell (MSC) osteogenesis by downregulating CAB39. This study reveals miRNA-451
Area of Science:
- Molecular Biology
- Stem Cell Biology
- Biochemistry
Background:
- Mesenchymal stem cells (MSCs) are crucial for bone regeneration.
- MicroRNAs (miRNAs) play regulatory roles in cell differentiation.
- The specific role of miRNA-451 in MSC osteogenesis requires further elucidation.
Purpose of the Study:
- To investigate the function of miRNA-451 in promoting MSC osteogenesis.
- To identify regulatory calcium binding protein 39 (CAB39) as a target of miRNA-451.
Main Methods:
- Dual-luciferase reporter assay to confirm miRNA-451 targeting of CAB39.
- Transfection of C3H10T1/2 cells with miRNA-451 mimics or inhibitors.
- Real-time PCR, Western blot, and Alizarin red staining to assess osteogenic markers and calcium deposition.
Main Results:
- miRNA-451 directly targets CAB39.
- Overexpression of miRNA-451 significantly increased Runx2 and ALP mRNA levels and extracellular calcium deposition.
- Inhibition of miRNA-451 decreased these osteogenic markers.
Conclusions:
- miRNA-451 promotes MSC osteogenesis by downregulating CAB39 expression.
- miRNA-451 is a potential therapeutic target for enhancing bone formation.
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