Related Experiment Video
Updated: Mar 27, 2026
Molecular Analysis of Endothelial-mesenchymal Transition Induced by Transforming Growth Factor-β Signaling
Published on: August 3, 2018
MicroRNA-483-3p Inhibits Extracellular Matrix Production by Targeting Smad4 in Human Trabecular Meshwork Cells
Wencui Shen1, Yating Han2, Bingqing Huang3
1Eye Hospital Eye Institute, Tianjin Medical University, Tianjin, China 2Department of Cell Biology, 2011 Collaborative Innovation Center of Tianjin for Medical Epigenetics, Laboratory of Epigenetics in Development and Tumorigenesis, Tianjin Research Cente.
Purpose:
This study investigated the effects of microRNA-483-3p (miR-483-3p) on extracellular matrix (ECM) production, and clarified the regulatory mechanism of microRNA-483-3p in human trabecular meshwork cells (HTMCs) under oxidative stress.
Methods:
The expression levels of ECM (fibronectin, laminin, collagen I) in HTMCs under oxidative stress were measured by Western blot. Changes of miR-483-3p expression in HTMCs were evaluated by quantitative polymerase chain reaction (qPCR). After using lentivirus stably expressing pri-miR-483, the effects of miR-483-3p on the ECM were assessed by qPCR and Western blot. Smad4, the potential target of miR-483-3p according to mRNA target-predicting algorithms, was confirmed by luciferase assay and Western blot. Furthermore, the effects of Smad4 knockdown on ECM expression were investigated by qPCR and Western blot.
Results:
The mRNA and protein levels of ECM (fibronectin, laminin, collagen I) were upregulated in HTMCs induced by oxidative stress. The expression level of miR-483-3p decreased in HTMCs under oxidative stress, and the ectopic expression of miR-483-3p decreased the levels of ECM. In addition, miR-483-3p targeted Smad4 through two binding sites, resulting in a decrease of Smad4 expression. Furthermore, knockdown of Smad4 reduced the levels of ECM in HTMCs.
Conclusions:
MicroRNA-483-3p has an inhibitory effect on ECM production in HTMCs through downregulating Smad4, which indicates that miR-483-3p may serve as a potential therapeutic target in glaucoma.
Insights
MicroRNA-483-3p inhibits extracellular matrix production in human trabecular meshwork cells by downregulating Smad4. This finding suggests miR-483-3p as a potential therapeutic target for glaucoma.
Area of Science:
- Ophthalmology
- Molecular Biology
- Cell Biology
Background:
- Oxidative stress is implicated in glaucoma pathogenesis.
- Extracellular matrix (ECM) production in trabecular meshwork cells (HTMCs) is crucial for aqueous humor outflow.
Purpose of the Study:
- To investigate the role of microRNA-483-3p (miR-483-3p) in regulating ECM production in HTMCs under oxidative stress.
- To elucidate the molecular mechanism underlying miR-483-3p's function.
Main Methods:
- Quantitative polymerase chain reaction (qPCR) and Western blot were used to assess ECM and miR-483-3p expression.
- Lentivirus-mediated overexpression of miR-483-3p and Smad4 knockdown were performed.
- Luciferase assay confirmed Smad4 as a direct target of miR-483-3p.
Main Results:
- Oxidative stress upregulated ECM components (fibronectin, laminin, collagen I) and downregulated miR-483-3p in HTMCs.
- Overexpression of miR-483-3p reduced ECM levels.
- miR-483-3p directly targeted and downregulated Smad4, which in turn reduced ECM production.
Conclusions:
- MicroRNA-483-3p inhibits ECM production in HTMCs by downregulating Smad4.
- miR-483-3p represents a potential therapeutic target for managing glaucoma.
More Related Videos
06:54Studying TGF-β Signaling and TGF-β-induced Epithelial-to-mesenchymal Transition in Breast Cancer and Normal Cells
Published on: October 27, 2020
07:05TGF-β-mediated Endothelial to Mesenchymal Transition EndMT and the Functional Assessment of EndMT Effectors using CRISPR/Cas9 Gene Editing
Published on: February 26, 2021
Related Concept Videos
01:16TGF - β Signaling Pathway
01:23Role of Matrix Metalloproteases in Degradation of ECM
01:29The Extracellular Matrix
In order to maintain tissue organization, many animal cells are surrounded by structural molecules that make up the extracellular matrix (ECM). Together, the molecules in the ECM maintain the structural integrity of tissue as well as the remarkable specific properties of certain tissues.
Composition of the Extracellular Matrix
The extracellular matrix (ECM) is commonly composed of ground substance, a gel-like fluid, fibrous components, and many structurally and functionally diverse...
01:42The Extracellular Matrix