MiR-424/TGIF2-Mediated Pro-Fibrogenic Responses in Oral Submucous Fibrosis
Abstract
:1. Introduction
2. Results
2.1. MiR-424 Is Aberrantly Overexpressed in OSF Tissues and Positively Associated with Several Fibrosis-Associated Molecules
2.2. Inhibition of MiR-424 in Fibrotic Buccal Mucosal Fibroblasts (fBMFs) Reduces Myofibroblast Activities
2.3. MiR-424 Is Associated with ROS Accumulation and Expression of Myofibroblast Marker
2.4. MiR-424 Modulates Myofibroblast Activities through Directly Binding to TGIF2
2.5. MiR-424 Induces Myofibroblast Transdifferentiation via Activation of TGF-β Signaling
3. Discussion
4. Materials and Methods
4.1. Tissue Collection, Cell Culture, and Reagents
4.2. Quantitative Real-Time PCR (qRT-PCR)
4.3. MiRNAs Mimic and Inhibition
4.4. Collagen Contraction Assay
4.5. Wound Healing Assay
4.6. Transwell Migration Assays
4.7. Reactive Oxygen Species Production Analysis
4.8. Western Blotting
4.9. MiRNA-Targeting Gene Prediction and Dual-Luciferase Reporter Assay
4.10. Lentiviral-Mediated RNAi for Silencing TGIF2:
4.11. ELISA Analysis
4.12. Statistical Analysis
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
Abbreviations
ACTA2 | Actin alpha 2 |
α-SMA | Alpha-smooth muscle actin |
BMFs | Buccal mucosal fibroblasts |
COL1A1 | Collagen type I alpha 1 chain |
DCF | 2′,7′-dichlorofluorescein |
DCFH-DA | 2′,7′- dichlorodihy-drofluorescein diacetate |
EMT | Epithelial-mesenchymal transition |
fBMF | Fibrotic buccal mucosal fibroblasts |
GAPDH | Glyceraldehyde-3-phosphate dehydrogenase |
MiR-424 | MicroRNA-424 |
OSF | Oral submucous fibrosis |
TGF-β1 | Transforming growth factor Beta 1 |
TGIF2 | TGFB-induced factor 2 protein |
ROS | Reactive oxygen species |
3′UTR | Three prime untranslated region |
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Chou, M.-Y.; Hsieh, P.-L.; Chao, S.-C.; Liao, Y.-W.; Yu, C.-C.; Tsai, C.-Y. MiR-424/TGIF2-Mediated Pro-Fibrogenic Responses in Oral Submucous Fibrosis. Int. J. Mol. Sci. 2023, 24, 5811. https://doi.org/10.3390/ijms24065811
Chou M-Y, Hsieh P-L, Chao S-C, Liao Y-W, Yu C-C, Tsai C-Y. MiR-424/TGIF2-Mediated Pro-Fibrogenic Responses in Oral Submucous Fibrosis. International Journal of Molecular Sciences. 2023; 24(6):5811. https://doi.org/10.3390/ijms24065811
Chicago/Turabian StyleChou, Ming-Yung, Pei-Ling Hsieh, Shih-Chi Chao, Yi-Wen Liao, Cheng-Chia Yu, and Chang-Yi Tsai. 2023. "MiR-424/TGIF2-Mediated Pro-Fibrogenic Responses in Oral Submucous Fibrosis" International Journal of Molecular Sciences 24, no. 6: 5811. https://doi.org/10.3390/ijms24065811