Phosphoglycerate kinase 1 silencing by a novel microRNA microRNA-4523 protects human osteoblasts from dexamethasone through activation of Nrf2 signaling cascade
| Autor: | Haining Tan, Lin Bo, Ming-sheng Tan, Jianhua Hu, Zhen-tao Zhou, Jinqian Liang |
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| Rok vydání: | 2021 |
| Předmět: |
Cancer Research
NF-E2-Related Factor 2 Immunology Down-Regulation Apoptosis Dexamethasone Article Cell Line Small hairpin RNA Cellular and Molecular Neuroscience Stress signalling Femur Head Necrosis microRNA medicine Gene silencing Humans Gene Silencing Phosphoglycerate kinase 1 education 3' Untranslated Regions Messenger RNA education.field_of_study Osteoblasts QH573-671 Base Sequence Chemistry Osteoblast Cell Biology Cell biology MicroRNAs Oxidative Stress Phosphoglycerate Kinase medicine.anatomical_structure GCLC Cytoprotection RNA Protein stabilization Cytology Signal Transduction |
| Zdroj: | Cell Death & Disease Cell Death and Disease, Vol 12, Iss 11, Pp 1-11 (2021) |
| ISSN: | 2041-4889 |
| Popis: | Nuclear-factor-E2-related factor 2 (Nrf2) cascade activation can ameliorate dexamethasone (DEX)-induced oxidative injury and death in human osteoblasts. Phosphoglycerate kinase 1 (PGK1) depletion is shown to efficiently activate Nrf2 signaling by inducing methylglyoxal modification of Kelch-like ECH-associated protein 1 (Keap1). We here identified a novel PGK1-targeting microRNA: microRNA-4523 (miR-4523). RNA fluorescent in situ hybridization, RNA pull-down, and Argonaute-2 RNA immunoprecipitation results confirmed a direct binding between miR-4523 and PGK1 mRNA in primary human osteoblasts and hFOB1.19 osteoblastic cells. Forced overexpression of miR-4523, using a lentiviral construct, robustly decreased PGK1 3′-UTR (untranslated region) luciferase activity and downregulated its expression in human osteoblasts and hFOB1.19 cells. Furthermore, miR-4523 overexpression activated the Nrf2 signaling cascade, causing Keap1–Nrf2 disassociation, Nrf2 protein stabilization, and its nuclear translocation as well as transcription activation of Nrf2-dependent genes (NQO1, GCLC, and HO1) in human osteoblasts. By expressing a UTR-null PGK1 construct, miR-4523 overexpression-induced Nrf2 cascade activation was however largely inhibited. Importantly, DEX-induced reactive oxygen species production, oxidative injury, and cell apoptosis were significantly attenuated by miR-4523 overexpression in human osteoblasts and hFOB1.19 cells. Such actions by miR-4523 were abolished by Nrf2 shRNA or knockout, but mimicked by PGK1 knockout (using CRISPR/Cas9 method). In PGK1 knockout human osteoblasts, miR-4523 overexpression failed to further increase Nrf2 cascade activation and offer osteoblast cytoprotection against DEX. Significantly, miR-4523 is downregulated in human necrotic femoral head tissues of DEX-taking patients. Together, PGK1 silencing by miR-4523 protected human osteoblasts from DEX through activation of the Nrf2 signaling cascade. |
| Databáze: | OpenAIRE |
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