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Published on: September 16, 2020
miR-155-5p regulates mesenchymal stem cell osteogenesis and proliferation by targeting GSK3B in steroid-associated
Fei Wu1, Wei Huang2, Yue Yang1
1Department of Orthopedics, Renmin Hospital, Wuhan University, Wuhan, Hubei, China.
Abstract:
microRNAs (miRNAs) have recently been recognized as playing an important role in bone-associated diseases. This study investigated whether the reduced miR-155-5p in steroid-associated osteonecrosis of the femoral head (ONFH) attenuated osteogenic differentiation and cell proliferation by targeting GSK3B. Bone marrow was collected from the proximal femurs of patients with steroid-associated ONFH (n = 10) and patients with new femoral neck fracture (n = 10) and mesenchymal stem cells (MSCs) were isolated. The expression profile, the biological function of miR-155-5p, and the interaction between miR-155-5p and GSK3B were investigated by cell viability measurement, western blot, real-time polymerase chain reaction, luciferase reporter assay, and Alizarin Red S (ARS) staining of MSCs. The MSCs that were obtained from the femoral neck fracture group and from the steroid-associated ONFH group were transfected with or without miR-155-5p. We found that, in ONFH samples, the level of mature miR-155-5p was significantly lower than that of control samples. By inhibiting GSK3B, miR-155-5p promoted the nuclear translocation of β-catenin, increased the expression of osteogenesis-related genes, and facilitated the proliferation and differentiation of MSCs. Restoring the expression of GSK3B in MSCs partially reversed the effect of miR-155-5p. These findings suggest that reduced miR-155-5p in steroid-associated ONFH attenuates osteogenic differentiation and cell proliferation by increased levels of GSK3B and inhibition of Wnt signaling.
Insights
Reduced miR-155-5p in steroid-associated osteonecrosis of the femoral head (ONFH) impairs bone cell growth and differentiation. This occurs by increasing GSK3B, which inhibits Wnt signaling crucial for bone formation.
Area of Science:
- Biomedical Science
- Molecular Biology
- Orthopedics
Background:
- MicroRNAs (miRNAs) are critical regulators in bone diseases.
- Steroid-associated osteonecrosis of the femoral head (ONFH) involves impaired bone metabolism.
Purpose of the Study:
- To investigate the role of miR-155-5p in steroid-associated ONFH.
- To determine if reduced miR-155-5p affects osteogenic differentiation and proliferation via GSK3B.
Main Methods:
- Mesenchymal stem cells (MSCs) isolated from ONFH and control patients.
- Analysis of miR-155-5p expression, cell proliferation, and osteogenic differentiation.
- Luciferase reporter assays to confirm GSK3B interaction.
Main Results:
- miR-155-5p levels were significantly lower in ONFH samples.
- miR-155-5p promoted MSC proliferation and osteogenic differentiation by inhibiting GSK3B.
- Inhibition of GSK3B led to β-catenin nuclear translocation and increased osteogenesis-related gene expression.
Conclusions:
- Reduced miR-155-5p in steroid-associated ONFH contributes to impaired osteogenesis.
- The mechanism involves elevated GSK3B levels and subsequent Wnt signaling inhibition.
- miR-155-5p may be a therapeutic target for steroid-associated ONFH.
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