Long noncoding RNA TUG1 inhibits osteogenesis of bone marrow mesenchymal stem cells via Smad5 after irradiation
| Autor: | Weiwei Zhang, Jiuxuan Li, Qian Ran, Fengjie Li, Lixin Xiang, Yang Xiang, Li Chen, Chun Wu, Xiaomei Zhang, Zhongjun Li, Jiang Wu |
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| Rok vydání: | 2019 |
| Předmět: |
Smad5 Protein
0301 basic medicine endocrine system animal structures Active Transport Cell Nucleus Medicine (miscellaneous) Bone Marrow Cells 03 medical and health sciences 0302 clinical medicine Downregulation and upregulation Western blot Osteogenesis medicine Humans Binding site Enhancer Pharmacology Toxicology and Pharmaceutics (miscellaneous) Cells Cultured Cell Nucleus medicine.diagnostic_test Microarray analysis techniques Chemistry Cell Differentiation Mesenchymal Stem Cells TUG1 Bone Marrow Mesenchymal Stem Cells In vitro Long non-coding RNA Cell biology 030104 developmental biology 030220 oncology & carcinogenesis embryonic structures Phosphorylation RNA Long Noncoding Irradiation Smad5 Research Paper |
| Zdroj: | Theranostics |
| ISSN: | 1838-7640 |
| Popis: | Irradiation can greatly inhibit osteogenesis of bone marrow mesenchymal stem cells (BM-MSCs). However, the mechanism remains unclear. Methods: We analyzed the expression profile of long noncoding RNAs (lncRNAs) in BM-MSCs using microarray data. LncRNA TUG1 (Taurine Upregulated Gene 1) was selected and tested in radiated BM-MSCs and non-radiated BM-MSCs. Functional analyses (in vitro) were performed to confirm the role of TUG1 in the osteogenic inhibition induced by irradiation. A RIP (RNA immunoprecipitation) assay was performed to detect the interaction of TUG1 and Smad5. Smad5 and the phosphorylated Smad5 (p-Smad5) were tested by western blot. The nuclear translocation of p-Smad5 were tested by immunofluorescence analysis. Furthermore, a series of Smad5 deletions was constructed to identify the TUG1 binding site of Smad5. Results: We found that numerous lncRNAs, including TUG1, exhibit significant expression differences after irradiation. After irradiation TUG1 was significantly increased in BM-MSCs and inhibited osteogenesis. Furthermore, TUG1 directly bound to Smad5, an osteogenic enhancer. Although the phosphorylation level of Smad5 was increased following irradiation, osteogenesis of BM-MSCs was decreased. Mechanistically, TUG1 interacting with the 50-90 aa region of Smad5 and blocks the nuclear translocation of p-Smad5, abolishing osteogenic signalling after irradiation. Conclusion: These results indicate that TUG1 is a negative regulator of Smad5 signalling and suppresses osteogenesis of BM-MSCs after irradiation. |
| Databáze: | OpenAIRE |
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