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Korean Society for Biochemistry and Molecular Biology (생화학분자생물학회)
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miR-101-3p/Rap1b signal pathway plays a key role in osteoclast differentiation after treatment with bisphosphonates

  • Li, Jie (Department of Orthopaedics, Xuzhou Central Hospital, Clinical School of Xuzhou Medical University) ;
  • Li, You (Department of Orthopaedics, The First Affiliated Hospital of Nanjing Medical University) ;
  • Wang, Shengjie (Department of Orthopedics Surgery, Henan Province People's Hospital) ;
  • Che, Hui (Department of Orthopaedics, The First Affiliated Hospital of Nanjing Medical University) ;
  • Wu, Jun (The Research Center for Bone and Stem Cells, Department of Anatomy, Histology and Embryology, Nanjing Medical University) ;
  • Ren, Yongxin (Department of Orthopaedics, The First Affiliated Hospital of Nanjing Medical University)
  • Received : 2019.03.19
  • Accepted : 2019.07.26
  • Published : 2019.09.30
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Abstract

Bisphosphonates are the mainstay of therapy worldwide for osteoporosis. However, bisphosphonates also have limitations. The objective of this study was to determine the role of miR-101-3p/Rap1b signal pathway in osteoclast differentiation after treatment with bisphosphonates. Our results revealed that miR-101-3p was an important regulator in bisphosphonates treated-osteoclasts. When miR-101-3p was down-regulated in bone marrow-derived macrophage-like cells (BMMs), the development of mature osteoclasts was promoted, and vice versa. However, alendronate decreased multinucleated cell number regardless of whether miR-101-3p was knocked down or over-expressed. TRAP activity assay confirmed the above results. Luciferase assay indicated that miR-101-3p was a negative regulator of Rap1b. Western blot analysis revealed that protein expression level of Rap1b in BMMs transfected with OV-miR-101-3p was lower than that in BMMs transfected with an empty vector. Rap1b overexpression increased TRAP-positive multinucleated cells, while Rap1b inhibition decreased the cell numbers. In vivo data showed that miR-101-3p inhibited osteoclast differentiation in ovariectomized mice while overexpressed of Rap1b blocked the differentiation. Taken together, our data demonstrate that miR-101-3p/Rap1b signal pathway plays a key role in osteoclast differentiation after treatment with bisphosphonates.

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References

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