| Popis: |
Human bone marrow mesenchymal stem cells (hBMSCs) have a strong self-renewal potential and osteogenic differentiation ability, thus providing a new method for bone defect repair research. LncRNA LINC00707 participates in the regulation of osteogenic differentiation of hBMSCs and our aim was to explore the potential regulatory mechanism.Firstly, quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression levels of LINC00707, miR-145, the low-density lipoprotein receptor-related protein 5 (LRP5) and osteogenesis-related genes. Next, alkaline phosphatase (ALP) activity assay was used to measure the relative activity of ALP in hBMSCs. The protein levels of LRP5 and osteogenesis-related genes were detected by Western blot. Finally, the relationship among LINC00707, miR-145 and LRP5 were predicted by online software and verified by Dual-Luciferase reporter assay, RNA pull-down and RNA immunoprecipitation (RIP).LINC00707 and osteogenesis-related genes were gradually upregulated during osteogenesis of hBMSCs. Meanwhile, overexpression of LINC00707 promoted osteogenic differentiation of hBMSCs. Interestingly, we found that LINC00707 negatively regulated the miR-145 expression and osteogenic differentiation functions by directly interacting with miR-145, and LINC00707 affected the functions of LRP5 by sponging miR-145 in hBMSCs. Moreover, LINC00707 promoted the Wnt/β-catenin pathway through the LINC00707/miR-145/LRP5 axis.LncRNA LINC00707 promoted osteogenic differentiation of hBMSCs by targeting LRP5 mediated by miR-145 through the activation of the Wnt/β-catenin pathway. |
