Abstract

Introduction

Tripartite motif 65 (TRIM65) is a crucial regulator of cell differentiation, proliferation, migration, invasion, and carcinogenesis. However, its role in osteoporosis (OP) remains unclear. In this study, we evaluated the role of TRIM65 in regulating osteoblast differentiation and calcification.

Materials and Methods

The role of TRIM65 during the osteogenic differentiation of MC3T3-E1 cells was evaluated. The expression of COL1A1, RUNX2, and OCN was examined using western blot analysis and immunofluorescence staining. The formation of calcium nodules was evaluated using alizarin red staining. Alkaline phosphatase activity was evaluated using ALP staining.

Results

TRIM65 expression was significantly elevated during the osteogenic differentiation of bone marrow mesenchymal stem and MC3T3-E1 cells. We demonstrated that TRIM65 overexpression enhanced osteogenic differentiation and promoted bone formation in the MC3T3-E1 cells. Conversely, TRIM65 inhibited the osteogenic differentiation and bone formation of the MC3T3-E1 cells. Mechanistically, we found that TRIM65 knockdown in MC3T3-E1 cells up-regulated the phosphorylated protein expression of PI3K and AKT, which was contrary to the results of the TRIM65-overexpression group.

Conclusion

Our research suggests that TRIM65 is an important osteogenic differentiation and bone formation regulator and offers a therapeutic application for OP.

Keywords: Tripartite motif 65, Osteoporosis, Osteoblast, Osteogenic differentiation, Signaling pathway, Phosphorylated protein.
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