P Fang1, L-X Zhang, Y Hu, L Zhang, L-W Zhou. 1. Department of Orthopedics, Jinling Hospital, Nanjing University, School of Medicine, Nanjing, China. LeiZHANG1987MD@163.com.
Abstract
OBJECTIVE: The aim of this study was to examine the role of lncRNA-differentiation antagonizing non-protein coding RNA (DANCR) and its underlying mechanisms in chondrogenesis, more specifically in synovial fluid-derived mesenchymal stem cell (SFMSCs). MATERIALS AND METHODS: The expression levels of DANCR in SFMSCs were measured by qRT-PCR. Luciferase reporter assay and RIP assay were used to investigate the direct target of DANCR and miR-1275 in SFMSCs. The expression of matrix metallopeptidase 13 (MMP13, also known as chondrogenic marker) protein was examined by Western blot. Cell proliferation was analyzed by Cell Counting Kit-8 (CCK-8) assay, while chondrogenic differentiation was explored by sGAG assay. RESULTS: Our data indicated that DANCR can promote SFMSCs proliferation and chondrogenesis. In addition, miR-1275 was indicated as a direct target of DANCR. MiR-1275 was negatively regulated by DANCR via competing endogenous RNA (ceRNA) mechanism. Moreover, our data revealed that miR-1275 could bind to MMP13 and regulate its expression. CONCLUSIONS: Our findings suggested that DANCR was involved in SFMSCs proliferation and chondrogenesis. Mechanistically, DANCR functions as a sponge RNA for miR-1275 that regulates the expression of target gene MMP13. These data provide a therapeutic option for Osteoarthritis (OA).
OBJECTIVE: The aim of this study was to examine the role of lncRNA-differentiation antagonizing non-protein coding RNA (DANCR) and its underlying mechanisms in chondrogenesis, more specifically in synovial fluid-derived mesenchymal stem cell (SFMSCs). MATERIALS AND METHODS: The expression levels of DANCR in SFMSCs were measured by qRT-PCR. Luciferase reporter assay and RIP assay were used to investigate the direct target of DANCR and miR-1275 in SFMSCs. The expression of matrix metallopeptidase 13 (MMP13, also known as chondrogenic marker) protein was examined by Western blot. Cell proliferation was analyzed by Cell Counting Kit-8 (CCK-8) assay, while chondrogenic differentiation was explored by sGAG assay. RESULTS: Our data indicated that DANCR can promote SFMSCs proliferation and chondrogenesis. In addition, miR-1275 was indicated as a direct target of DANCR. MiR-1275 was negatively regulated by DANCR via competing endogenous RNA (ceRNA) mechanism. Moreover, our data revealed that miR-1275 could bind to MMP13 and regulate its expression. CONCLUSIONS: Our findings suggested that DANCR was involved in SFMSCs proliferation and chondrogenesis. Mechanistically, DANCR functions as a sponge RNA for miR-1275 that regulates the expression of target gene MMP13. These data provide a therapeutic option for Osteoarthritis (OA).