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Erschienen in:
02.03.2020 | Original Paper
Silencing of p53 reduces cell migration in human Tenon's fibroblasts induced by TGF-β
Erschienen in: International Ophthalmology | Ausgabe 6/2020
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Purpose
Growth factors are considered as key molecules that participating in fibrosis formation. This research aimed to clarify potential effects of p53 on regulation of transforming growth factor β (TGF-β) and fibrosis formation and investigate the associated mechanisms.
Methods
Vimentin was examined to identify human Tenon's fibroblasts (HTFs). p53-targeting small interfere RNA (siRNA) was synthesis and transfected into HTFs. Real-time PCR assay was utilized to evaluate p53 and microRNA-29b (miR-29b) expression. Immunocytochemical assay was used to observe TGF-β expression. The wound healing assay was conducted to evaluate migration of HTFs. Dual-luciferase assay was employed to identify interaction between p53 and miR-29b in HTFs.
Results
Vimentin was extensively distributed in HTFs cells. HTFs at density of 5 × 104 cells/ml and 6 days exhibited the best growth. The p53 level in TGF-β treatment group was significantly higher compared to that in blank group (p < 0.01). miR-29b level in siRNA targeting p53 group was significantly increased compared to that in blank group (p < 0.01). siRNA targeting p53 could significantly inhibit HTFs migration compared to that in single TGF-β treating HTFs group (p < 0.01). Relative luciferase activity was significantly increased in p53 overexpressed HTFs compared to that in cells transfected with empty pcDNA3.0 plasmid (p < 0.01).
Conclusions
p53 inhibited expression of TGF-β, suppressed HTFs migration and inhibited HTFs growth, by reducing miR-29b expression and interacting with miR29b gene in HTFs.