Volume 29, Issue 3
DOI: 10.24205/03276716.2020.740
Research on the Mechanism of miR-142-5p’s Promotion of the Function and Mineralization of Osteoblasts by Down-regulating the Expression of WWP1 Gene
Abstract
Objective: To study the mechanism of miR-142-5p’s promotion of the function and mineralization of osteoblasts by down-regulating the expression of WWP1 gene.
Methods:a fracture model of wild-type SPF mice was established, and the expression of miR-142-5p in callus tissues of mice was detected by real-time quantitative PCR. The osteoblastic cell line SV40 was cultured in vitro. The SV40 cells were treated with agonistmiR142-5p, agonist control, antagonist-miR-142-5p and antagonist control reagents. miR-142-5pacquired and miR-142-5p-deficientSV40 cell modelswere constructed, and a blank control group was established. miR-142-5p was transfected respectively, and its effects on ALP mRNA, OC mRNA, matrix mineralization and mRNA and protein of the target gene WWP1 were analyzed.
Result:The levels of miR-142-5p in callus tissues on 7, 14, 21 and 28d were detected after modeling by quantitative PCR in real time and the results showed that miR-142-5p expression was significantly enabled during fracture healing, peaked at 21d after modeling, and marginally decreased at 28d. In the agonist miR-142-5p group, the levels of ALP mRNA, OC mRNA, and alizarin red S (ARS) were meaningfully higher than in the agonist control group, and The levels in the antagonist-miR-142-5p group for ALP mRNA, OC mRNA and ARS quantification were substantially lower than those in the antagonist control group. There was no substantial difference in WWP1 mRNA expression among four groups, the expression of WWP1 protein in the agonist miR-142-5p group was significantly lower than in the agonist control group; the expression of WWP1 protein in the antagonist-miR-142-5p group was meaningfully higher than in the antagonist control group.
Conclusion: the activation of the expression of miR-142-5p in callus tissues during fracture healing is closely related to the activity of osteoblasts, and supports the function and
mineralization of osteoblasts. The mechanism may be related to miR-142-5p’s downregulation of EEP1 gene at the post-transcriptional level.l
Keywords
miR-142-5p, WWP1 gene, Osteoblast, Mineralization, Mechanism.