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5-Aza-2′-deoxycytidine induces human Tenon's capsule fibroblasts differentiation and fibrosis by up-regulating TGF-β type I receptor
5-Aza-2′-deoxycytidine induces human Tenon's capsule fibroblasts differentiation and fibrosis by up-regulating TGF-β type I receptor
- Source :
- Experimental Eye Research. 165:47-58
- Publication Year :
- 2017
- Publisher :
- Elsevier BV, 2017.
-
Abstract
- The principle reason of high failure rate of glaucoma filtration surgery is the loss of filtration function caused by postoperative scar formation. We investigated the effects of 5-aza-2′-deoxycytidine (5-Aza-dc), a DNA methyltransferases inhibitor, on human Tenon's capsule fibroblasts (HTFs) differentiation and fibrosis and its mechanism of action, especially in relation to transforming growth factor (TGF)-β1 signaling. TGF-β1 was used to induce differentiation of cultured HTFs. 5-Aza-dc suppressed DNA methyltransferases (DNMTs) activity 6 h after treatment with a course corresponding to that of TGF-β1-induced reduction of DNMT activity without affecting cell viability as measured by Cell Counting Kit-8 assay. 5-Aza-dc also reduced DNMT1 and DNMT3a protein expression from 24 to 48 h. HTFs migration evaluated by scratch-wound assay were significantly increased 24 h after 5-Aza-dc treatment, a time course similar to that of TGF-β1. Treatment with 5-Aza-dc significantly increased the mRNA and protein levels of α-smooth muscle actin (α-SMA), collagen-1A1 (Col1A1), fibronectin (FN) and TGF-β type I receptor (TGFβRI). Furthermore, the effects of 5-Aza-dc on DNMT activity suppression, cell migration, and fibrosis were all reversed by a TGFβRI inhibitor- SB-431542. Meanwhile, knockdown of DNMT1 upregulated TGFβRI expression and had the same fibrosis-inducing effect in HTFs, which was also inhibited by SB-431542. Thus, the results indicate that DNA hypomethylation induces HTFs differentiation and fibrosis through up-regulation of TGFβRI. DNA methylation status plays an important role in subconjunctival wound healing.
- Subjects :
- Adult
DNA (Cytosine-5-)-Methyltransferase 1
Male
0301 basic medicine
Methyltransferase
Tenon Capsule
Receptor, Transforming Growth Factor-beta Type I
Cell Count
Protein Serine-Threonine Kinases
Decitabine
DNA Methyltransferase 3A
03 medical and health sciences
Cellular and Molecular Neuroscience
0302 clinical medicine
Tenon's capsule
Transforming Growth Factor beta
Fibrosis
medicine
Humans
DNA (Cytosine-5-)-Methyltransferases
Viability assay
Enzyme Inhibitors
Cells, Cultured
Gene knockdown
biology
Cell Differentiation
Cell migration
DNA Methylation
Fibroblasts
Middle Aged
medicine.disease
Molecular biology
Sensory Systems
Up-Regulation
Repressor Proteins
Fibronectin
Ophthalmology
030104 developmental biology
medicine.anatomical_structure
Azacitidine
030221 ophthalmology & optometry
Cancer research
biology.protein
Female
Receptors, Transforming Growth Factor beta
Transforming growth factor
Subjects
Details
- ISSN :
- 00144835
- Volume :
- 165
- Database :
- OpenAIRE
- Journal :
- Experimental Eye Research
- Accession number :
- edsair.doi.dedup.....e2a441c5c8b42f4f8927da0e92e2dabe
- Full Text :
- https://doi.org/10.1016/j.exer.2017.09.004