W-L Cai1, W Zeng, H-H Liu, B-Y Zhu, J-L Liu, Y Liu. 1. Department of Orthopedics Surgery, The Second Xiangya Hospital of Central South University, Changsha, China. zengwei198108@163.com.
Abstract
OBJECTIVE: 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. MATERIALS AND METHODS: 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). RESULTS: 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. CONCLUSIONS: LncRNA LINC00707 promoted osteogenic differentiation of hBMSCs by targeting LRP5 mediated by miR-145 through the activation of the Wnt/β-catenin pathway.
OBJECTIVE: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. MATERIALS AND METHODS: 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). RESULTS: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. CONCLUSIONS: LncRNA LINC00707 promoted osteogenic differentiation of hBMSCs by targeting LRP5 mediated by miR-145 through the activation of the Wnt/β-catenin pathway.
Authors: Carmen Lanzillotti; Monica De Mattei; Chiara Mazziotta; Francesca Taraballi; John Charles Rotondo; Mauro Tognon; Fernanda Martini Journal: Front Cell Dev Biol Date: 2021-04-09