LncRNA XIST facilitates S1P-mediated osteoclast differentiation via interacting with FUS
| Autor: | Da-Wei Zhang, Hong-Gang Wang, Kui-Bo Zhang, Yuan-Qing Guo, Lian-Jun Yang, Hai Lv |
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| Rok vydání: | 2021 |
| Předmět: |
NFATC Transcription Factors
Tartrate-Resistant Acid Phosphatase Endocrinology Diabetes and Metabolism Cathepsin K RANK Ligand Serine Endopeptidases Osteoclasts Cell Differentiation General Medicine Hematopoiesis Mice Phosphotransferases (Alcohol Group Acceptor) Endocrinology RAW 264.7 Cells Osteogenesis Animals RNA-Binding Protein FUS Orthopedics and Sports Medicine RNA Long Noncoding Proprotein Convertases Bone Resorption |
| Zdroj: | Journal of bone and mineral metabolism. 40(2) |
| ISSN: | 1435-5604 |
| Popis: | The diagnosis and treatment of osteoporosis, a frequent age-related metabolic bone disorder, remain incomprehensive and challenging. The potential regulatory role of lncRNA XIST and sphingosine kinase 1 (SPHK1) pathway need experimental investigations.RAW264.7 cells and BMMs were obtained for in vitro studies and 30 ng/mL RANKL was implemented for induction of osteoclast differentiation. The suppressing of lncRNA XIST, SPHK1 and fused in sarcoma (FUS) was achieved using small hairpin RNA, while overexpression of XIST and FUS was constructed by pcDNA3.1 vector system. Tartrate-resistant acid phosphatase (TRAP) staining was used for observation of formation of osteoclasts. RNA-pulldown analysis and RNA binding protein immunoprecipitation (RIP) was implemented for measuring mRNA and protein interactions. RT-qPCR was conducted to determining mRNA expression, whereas ELISA and Western blotting assay was performed for monitoring protein expression.RANKL induced osteoclast differentiation and upregulated expression of osteoclastogenesis-related genes that included NFATc1, CTSK, TRAP and SPHK1 and the level of lncRNA XIST in both RAW264.7 cells and BMMs. However, knockdown of lncRNA XIST or suppressing SPHK1 significantly reserved the effects of RANKL. LncRNA XIST was further demonstrated to be interacted with FUS and increased the stability of SPHK1, indicating its ability in promoting osteoclast differentiation through SPHK1/S1P/ERK signaling pathway.LncRNA XIST promoted osteoclast differentiation via interacting with FUS and upregulating SPHK1/S1P/ERK pathway. |
| Databáze: | OpenAIRE |
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