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Published on: August 3, 2018
MiR-151b inhibits osteoblast differentiation via downregulating Msx2
Fuan Liu1, Yunbang Liang1, Xiaoyi Lin1
1Department of Orthopedics, The First Affiliated Hospital of Xiamen University, Xiamen, People's Republic of China.
Purpose:
MicroRNA-151b (miR-151b) showed altered expression in ovariectomized rat model of osteoporosis. This study established an ovariectomy-induced osteoporotic rat model to investigate the role of miR-151b in osteoblasts.
Methods:
Eighteen female Sprague-Dawley (SD) rats were divided randomly into Sham and OVX group (n = 9). The transfections with different miRNAs and expression vectors were confirmed by RT-qPCR. The protein expression of Msx2 was detected by Western blots. The interaction between miR-151b and Msx2D was evaluated by RNA pull-down and dual luciferase reporter assay.
Results:
The expression of miR-151b was significantly increased in femoral tissues of ovariectomy-induced osteoporotic rats. The expression of osteogenesis marker genes including RUNX2, ALP, OCN, OSX, and Msx2 were all significantly increased in osteogenic medium (OM) incubated primary osteoblasts and MC3T3-E1 cells. The interaction between miR-151b and Msx2 was confirmed by luciferase reporter assay and RNA pull-down. Moreover, overexpression of miR-151b significantly inhibited Msx2 in both MC3T3-E1 cells and primary osteoblasts, while miR-151b inhibitor had the opposite effect on the expression of Msx2. In addition, in primary osteoblasts and MC3T3-E1 cells, miR-151b overexpression, or Msx2 silence significantly decreased the expression of OSX, ALP, RUNX2, and OCN.
Conclusion:
MiR-151b could inhibit osteoblast proliferation, differentiation, and mineralization via downregulating Msx2 in both MC3T3-E1 cells and primary osteoblasts. MiR-151b might serve as a novel therapeutic target for osteoporosis.
Abbreviations:
miR-151b: microRNA-151b; miRNAs: microRNAs; Msx2: Msh homeobox 2; MAPK: mitogen-activated protein kinase; STAT: signal transducer and activator of transcription; SD: Sprague-Dawley; BMD: bone mineral density; qRT-PCR: quantitative reverse transcription PCR; MTT: methyl thiazolyl tetrazolium; OVX: ovariectomy; ALP: alkaline phosphatase.
Insights
MicroRNA-151b (miR-151b) inhibits osteoblast function by downregulating Msx2, suggesting it as a therapeutic target for osteoporosis. This study investigated miR-151b
Area of Science:
- Molecular Biology
- Cell Biology
- Biochemistry
Background:
- Osteoporosis is a bone disease characterized by low bone mass and microarchitectural deterioration.
- MicroRNAs (miRNAs) play crucial roles in regulating bone metabolism.
- MicroRNA-151b (miR-151b) has shown altered expression in osteoporosis models.
Purpose of the Study:
- To investigate the role of miR-151b in osteoblasts using an ovariectomy-induced osteoporotic rat model.
- To elucidate the molecular mechanism by which miR-151b affects osteoblast function.
- To explore miR-151b as a potential therapeutic target for osteoporosis.
Main Methods:
- An ovariectomy (OVX)-induced osteoporotic rat model was established.
- Transfection of cells with miRNAs and expression vectors, confirmed by RT-qPCR.
- Western blot analysis for Msx2 protein expression.
- RNA pull-down and dual luciferase reporter assays to confirm interaction between miR-151b and Msx2.
Main Results:
- miR-151b expression was significantly increased in osteoporotic rat femoral tissues.
- Overexpression of miR-151b inhibited Msx2 expression in osteoblasts.
- miR-151b overexpression or Msx2 silence decreased the expression of osteogenesis markers (OSX, ALP, RUNX2, OCN).
Conclusions:
- MiR-151b inhibits osteoblast proliferation, differentiation, and mineralization by downregulating Msx2.
- MiR-151b represents a potential novel therapeutic target for osteoporosis treatment.
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