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miR-101-3p/Rap1b signal pathway played a key role in osteoclast differentiation after treatment with bisphosphonates
Jie Li1,#, You Li2,#, Shengjie Wang3, Hui Che2, Jun Wu4,*, Yongxin Ren2,*
1Department of Orthopaedics, Xuzhou Central Hospital, Xuzhou Clinical School of Xuzhou Medical University, The Affiliated XuZhou Hospital of Medical College of Southeast University, Xuzhou Clinical Medical College of Nanjing Uni,
2Department of Orthopedics, The First Affiliated Hospital of Nanjing Medical University,
3Department of Orthopaedics Surgery, The Henan Province People’s Hospital,
4Department of Anatomy, Histology and Embryology, The Research Center for Bone and Stem Cells, Nanjing Medical University
Abstract
Bisphosphonates are the mainstay of therapy worldwide for osteoporosis. However, bisphosphonates still have their limitations. Our study implied miR-101-3p is a important regulator in bisphosphonates treated-osteoclasts. When miR-101-3p were down-regulated in bone marrow-derived macrophage-like cells (BMMs), the development of mature osteoclasts were promoted, and vice versa. However, alendronate can decrease the multinucleated cell whether miR-101-3p is knock-down or over-expression. TRAP activity assay also confirmed above results.Luciferase assay indicated that miR-101-3p is a negative regulator of Rap1b and western blot analysis displayed that protein expression level of Rap1b in BMMs transfected with OV-miR-101-3p was lower than in BMMs transfected with empty vector. And Rap1b overexpression can increase the TRAP-positive multinucleated cells, and vice versa. Furthermore, in vivo data showed that miR-101-3p could inhibit osteoclast differentiation in ovariectomized mice,while overexpressed Rap1b can block it. Our data demonstrate that miR-101-3p/Rap1b signal pathway played a key role in osteoclast differentiation after treatment with bisphosphonates.
Abstract, Accepted Manuscript(in press) [Submitted on March 19, 2019, Accepted on July 26, 2019]
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