Article ; Online: DCAF1-targeting microRNA-3175 activates Nrf2 signaling and inhibits dexamethasone-induced oxidative injury in human osteoblasts.
2021 Volume 12, Issue 11, Page(s) 1024
Abstract: Activation of nuclear-factor-E2-related factor 2 (Nrf2) signaling can protect human osteoblasts from dexamethasone-induced oxidative injury. DDB1 and CUL4 associated factor 1 (DCAF1) is a novel ubiquitin E3 ligase for Nrf2 protein degradation. We ... ...
| Abstract | Activation of nuclear-factor-E2-related factor 2 (Nrf2) signaling can protect human osteoblasts from dexamethasone-induced oxidative injury. DDB1 and CUL4 associated factor 1 (DCAF1) is a novel ubiquitin E3 ligase for Nrf2 protein degradation. We identified a novel DCAF1-targeting miRNA, miR-3175. RNA pull-down, Argonaute 2 RNA-immunoprecipitation, and RNA fluorescent in situ hybridization results confirmed a direct binding between miR-3175 and DCAF1 mRNA in primary human osteoblasts. DCAF1 3'-untranslated region luciferase activity and its expression were significantly decreased after miR-3175 overexpression but were augmented with miR-3175 inhibition in human osteoblasts and hFOB1.19 osteoblastic cells. miR-3175 overexpression activated Nrf2 signaling, causing Nrf2 protein stabilization, antioxidant response (ARE) activity increase, and transcription activation of Nrf2-dependent genes in human osteoblasts and hFOB1.19 cells. Furthermore, dexamethasone-induced oxidative injury and apoptosis were largely attenuated by miR-3175 overexpression in human osteoblasts and hFOB1.19 cells. Importantly, shRNA-induced silencing or CRISPR/Cas9-mediated Nrf2 knockout abolished miR-3175 overexpression-induced osteoblast cytoprotection against dexamethasone. Conversely, DFAC1 knockout, by the CRISPR/Cas9 method, activated the Nrf2 cascade and inhibited dexamethasone-induced cytotoxicity in hFOB1.19 cells. Importantly, miR-3175 expression was decreased in necrotic femoral head tissues of dexamethasone-taking patients, where DCAF1 mRNA was upregulated. Together, silencing DCAF1 by miR-3175 activated Nrf2 signaling to inhibit dexamethasone-induced oxidative injury and apoptosis in human osteoblasts. |
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| MeSH term(s) | Apoptosis/genetics ; Case-Control Studies ; Dexamethasone/pharmacology ; Femur Head/drug effects ; Femur Head/metabolism ; Femur Head/pathology ; Gene Knockout Techniques ; Gene Silencing ; HEK293 Cells ; Humans ; MicroRNAs/genetics ; MicroRNAs/metabolism ; NF-E2-Related Factor 2/genetics ; NF-E2-Related Factor 2/metabolism ; Necrosis ; Osteoblasts/drug effects ; Osteoblasts/metabolism ; Oxidative Stress/drug effects ; Protein Binding ; Protein Serine-Threonine Kinases/genetics ; Protein Serine-Threonine Kinases/metabolism ; RNA, Messenger/genetics ; Reactive Oxygen Species/metabolism ; Signal Transduction/genetics ; Transfection ; Ubiquitin-Protein Ligases/genetics ; Ubiquitin-Protein Ligases/metabolism |
| Chemical Substances | MIRN3175 microRNA, human ; MicroRNAs ; NF-E2-Related Factor 2 ; NFE2L2 protein, human ; RNA, Messenger ; Reactive Oxygen Species ; Dexamethasone (7S5I7G3JQL) ; Ubiquitin-Protein Ligases (EC 2.3.2.27) ; DCAF1 protein, human (EC 2.7.11.1) ; Protein Serine-Threonine Kinases (EC 2.7.11.1) |
| Language | English |
| Publishing date | 2021-10-29 |
| Publishing country | England |
| Document type | Journal Article ; Research Support, Non-U.S. Gov't |
| ZDB-ID | 2541626-1 |
| ISSN | 2041-4889 ; 2041-4889 |
| ISSN (online) | 2041-4889 |
| ISSN | 2041-4889 |
| DOI | 10.1038/s41419-021-04300-8 |
| Database | MEDical Literature Analysis and Retrieval System OnLINE |
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