Original Article
Effect of miR-204 on proliferation and differentiation of osteoblasts in osteoporosis mice by Wnt signaling pathway and its mechanism
Luo Jian, He Rongzhen, He Xingwen, Yang Chunxi, Qiu Huibin
Published 2020-02-25
Cite as Chin J Endocr Surg, 2020,14(01): 60-66. DOI: 10.3760/cma.j.issn.1674-6090.2020.01.014
Abstract
ObjectiveTo investigate the effect of miR-204 on the proliferation and differentiation of osteoblasts in osteoporosis mice by Wnt signaling pathway and its mechanism.
MethodsFemale Kunming mice were divided into: control group, sham operation group and osteoporosis group. Ovariectomy mouse models were established and identified by bilateral ovariectomy; Mouse primary osteoblasts were extracted and identified; Cells was transfected and detected the miR-204 expression levels; MTT was used to detect the viability of each group of cells; Alkaline phosphatase (ALP) activity was detected in each cell group; Cell flowmetry was used to detect apoptosis in each group; Cell flowmetry was used to detect the activity of Caspase-3 in each group of cells; Interaction between miR-204, β-catenin and LRP-5 was detected by dual luciferase reporter gene. Western blot was used to detect the expression of Wnt signaling pathway-related proteins.
ResultsThe bone mineral density of the osteoporosis group was significantly lower than that of the control group and the sham operation group (P=0.007, P=0.057) , indicating that the osteoporosis mice were successfully modeled; The expression level of miR-204 was significantly increased in the miR-204 mimics group (P=0.007) , and decreased in the miR-204 inhibitor group (P=0.031) ; The activity of bone cell and ALP activity of miR-204 mimics increased (P=0.007, P=0.043) , and the activity of bone cell and ALP decreased by miR-204 inhibitor (P=0.007, P=0.035) ; The invasive ability of miR-204 mimics was significantly increased (P=0.006) , and the invasive ability of miR-204 inhibitor was decreased (P=0.036) ; The apoptosis ability and Caspase-3 activity of miR-204 mimics were decreased (P=0.041, P=0.045) , and the apoptosis ability and Caspase-3 activity of bone cells were enhanced by miR-204 inhibitor (P=0.005, P=0.039) ; There were targeting relationship between miR-204 and β-catenin, LRP-5. The expressions of β-catenin and LRP-5 protein in osteoblasts of miR-204 mimics were up-regulated (P=0.043, P=0.009) , and the expression of β-catenin and LRP-5 protein in bone cells of miR-204 inhibitor was down-regulated (P=0.041, P=0.032) .
ConclusionmiR-204 maybe promote the proliferation and differentiation of osteoblasts, activate Wnt signaling pathway, and has certain protective effect on osteoporosis.
Key words:
miR-204; Wnt signaling pathway; Osteoporosis mice; Osteoblasts; Proliferation
Contributor Information
Luo Jian
Ningbo Hangzhou Bay Hospital, Department of Orthopaedics, Ningbo 315300, China
He Rongzhen
Ningbo Hangzhou Bay Hospital, Department of Orthopaedics, Ningbo 315300, China
He Xingwen
Ningbo Hangzhou Bay Hospital, Department of Orthopaedics, Ningbo 315300, China
Yang Chunxi
Department of Joint Surgery, Renji Hospital, Shanghai Jiao Tong University Shanghai 200001, China
Qiu Huibin
Department of Orthopaedics, Ningbo Second Hospital, Ningbo 315502, China
