Related Experiment Video
Updated: Jan 29, 2026
Promoter Capture Hi-C: High-resolution, Genome-wide Profiling of Promoter Interactions
Published on: June 28, 2018
MicroRNA-9-5p promotes osteoporosis development through inhibiting osteogenesis and promoting adipogenesis via
1Department of Traumatology, Yantaishan Hospital, Yantai, China. fanba05328829@163.com.
Objective:
To explore the role of microRNA-9-5p in regulating osteoporosis (OS) development and its underlying mechanism.
Patients And Methods:
MicroRNA-9-5p expression in peripheral blood of 30 OS patients and 30 healthy subjects was examined by quantitative Real-Time-Polymerase Chain Reaction (qRT-PCR). During the processes of osteogenesis and adipogenesis, mRNA levels of microRNA-9-5p, osteogenesis-related genes, and adipogenesis-related genes in marrow stromal stem cells (MSCs) were detected by qRT-PCR as well. After overexpression or knockdown of microRNA-9-5p, the regulatory effects of microRNA-9-5p on osteogenesis-related genes and adipogenesis-related genes in MSCs were accessed by detecting their mRNA and protein levels. Alizarin red staining and oil red staining were performed to determine the osteogenic and adipogenic capacities of MSCs after microRNA-9-5p overexpression, respectively. The dual-luciferase reporter gene assay was conducted to verify the binding condition of microRNA-9-5p and Wnt3a. Finally, rescue experiments were performed to confirm whether microRNA-9-5p could regulate OS development via targeting Wnt3a.
Results:
Higher expression of microRNA-9-5p was found in OS patients than that of healthy controls. MicroRNA-9-5p expression was downregulated with the prolongation of osteogenic induction, whereas it was upregulated during the process of adipogenic differentiation. Overexpression of microRNA-9-5p downregulated mRNA levels of osteogenesis-related genes (ALP, RUNX2, and OPN), whereas upregulated adipogenesis-related genes (PPARγ, Adipsin, and C/EBPα) in MSCs. The number of calcified nodules became fewer after microRNA-9-5p overexpression in MSCs. MSCs that overexpressed microRNA-9-5p showed more lipid droplets than that of controls. Subsequently, the dual-luciferase reporter gene assay verified that Wnt3a is the target gene of microRNA-9-5p. Both mRNA and protein levels of Wnt3a were negatively regulated by microRNA-9-5p. Rescue experiments indicated that the regulatory effects of microRNA-9-5p on osteogenesis and adipogenesis of MSCs were reversed by Wnt3a overexpression.
Conclusions:
MicroRNA-9-5p is lowly expressed in the peripheral blood of OS patients. MicroRNA-9-5p promotes the occurrence and progression of OS through inhibiting osteogenesis and promoting adipogenesis via targeting Wnt3a.
Insights
MicroRNA-9-5p is downregulated in osteoporosis patients and promotes the condition by inhibiting bone formation and enhancing fat cell differentiation via Wnt3a signaling. This study reveals microRNA-9-5p as a potential therapeutic target for osteoporosis.
Area of Science:
- Molecular Biology
- Biochemistry
- Genetics
Background:
- Osteoporosis (OS) is a metabolic bone disease characterized by decreased bone mineral density and increased fracture risk.
- MicroRNAs (miRNAs) are small non-coding RNAs that regulate gene expression and play critical roles in various biological processes, including bone metabolism.
Purpose of the Study:
- To investigate the role of microRNA-9-5p in the development of osteoporosis.
- To elucidate the underlying molecular mechanism of microRNA-9-55p in regulating osteogenesis and adipogenesis.
Main Methods:
- Quantitative Real-Time-Polymerase Chain Reaction (qRT-PCR) to measure microRNA-9-5p expression in patients and cells.
- In vitro studies involving overexpression and knockdown of microRNA-9-5p in marrow stromal stem cells (MSCs).
- Alizarin red and oil red staining to assess osteogenic and adipogenic differentiation.
- Dual-luciferase reporter gene assay to confirm Wnt3a as a target gene.
Main Results:
- MicroRNA-9-5p expression was significantly higher in OS patients compared to healthy controls and was inversely correlated with osteogenic differentiation while positively correlated with adipogenic differentiation in MSCs.
- Overexpression of microRNA-9-5p inhibited osteogenesis and promoted adipogenesis in MSCs, evidenced by reduced calcified nodules and increased lipid droplets.
- Wnt3a was identified as a direct target of microRNA-9-5p, with microRNA-9-5p negatively regulating Wnt3a expression at both mRNA and protein levels.
- Wnt3a overexpression reversed the effects of microRNA-9-5p on osteogenesis and adipogenesis.
Conclusions:
- MicroRNA-9-5p is downregulated in osteoporosis patients.
- MicroRNA-9-5p promotes osteoporosis development by inhibiting osteogenesis and promoting adipogenesis through the Wnt3a signaling pathway.
- MicroRNA-9-5p represents a potential therapeutic target for osteoporosis treatment.
More Related Videos
10:58Plant Promoter Analysis: Identification and Characterization of Root Nodule Specific Promoter in the Common Bean
Published on: December 23, 2017
06:51Systemic Delivery of MicroRNA Using Recombinant Adeno-associated Virus Serotype 9 to Treat Neuromuscular Diseases in Rodents
Published on: August 10, 2018
Related Concept Videos
02:40The Eukaryotic Promoter Region
02:40The Eukaryotic Promoter Region
00:50Anaphase Promoting Complex
01:18Models of Health Promotion and Illness Prevention II
The agent-host-environment model states that disease results...
01:22MicroRNAs
01:26Levels of Health Promotion and Illness Prevention
In primary prevention, actions taken before disease onset prevent the disease from...