LncRNA MSC-AS1 promotes osteogenic differentiation and alleviates osteoporosis through sponging microRNA-140–5p to upregulate BMP2
| Autor: | Zhihui Huang, Shuanghua He, Xinyu Hu, Feng Wang, Yiwen Zhao, Naidong Zhang, Wenge Ding |
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| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
Biophysics Bone Morphogenetic Protein 2 Biochemistry Bone morphogenetic protein 2 03 medical and health sciences 0302 clinical medicine Western blot Downregulation and upregulation Osteogenesis microRNA medicine Humans Molecular Biology Cells Cultured Reporter gene Gene knockdown medicine.diagnostic_test Chemistry Cell Biology Up-Regulation Cell biology RUNX2 MicroRNAs 030104 developmental biology medicine.anatomical_structure 030220 oncology & carcinogenesis embryonic structures Osteoporosis RNA Long Noncoding Bone marrow |
| Zdroj: | Biochemical and Biophysical Research Communications. 519:790-796 |
| ISSN: | 0006-291X |
| DOI: | 10.1016/j.bbrc.2019.09.058 |
| Popis: | This study aims to explore the role of lncRNA MSC-AS1/microRNA-140-5p/BMP2 regulatory loop in promoting osteogenic differentiation of BMSCs. BMSCs were isolated from bone marrow of mice. Expression levels of MSC-AS1, microRNA-140-5p and BMP2 during osteogenic differentiation were detected by qRT-PCR. Meanwhile, regulatory effect of MSC-AS1 on osteogenic differentiation was detected through ALP staining and alizarin red staining. The binding sites between microRNA-140-5p and MSC-AS1 as well as between microRNA-140-5p and BMP2 were predicted by TargetScan, which were further confirmed by dual-luciferase reporter gene assay. In addition, protein levels of MSC-AS1/microRNA-140-5p/BMP2 were detected by Western blot. Finally, rescue experiments were conducted to clarify the regulatory effects of MSC-AS1/microRNA-140-5p/BMP2 axis on osteogenic differentiation. MSC-AS1 and BMP2 were found to be remarkably up-regulated during osteogenic differentiation, while microRNA-140-5p was conversely down-regulated. Meanwhile, knockdown of MSC-AS down-regulated expression levels of osteogenesis-associated genes and weakened the mineralization capacity of BMSCs. MicroRNA-140-5p was verified to bind to the 3'UTR of MSC-AS1 and BMP2 genes. Knockdown of MSC-AS1 in BMSCs could reduce the expression of microRNA-140-5p, while knockdown of microRNA-140-5p also down-regulated BMP2 level. In addition, co-silence of MSC-AS1 and microRNA-140-5p reversed the inhibitory effect of MSC-AS1 knockdown on osteogenic differentiation and protein levels of p-Smad1/5/8, RUNX2 and Osterix. MSC-AS1 might promote the osteogenic differentiation of BMSCs through sponging microRNA-140-5p to up-regulate BMP2, thus alleviating the progression of osteoporosis. |
| Databáze: | OpenAIRE |
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