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MiR-199a-3p/5p participated in TGF-β and EGF induced EMT by targeting DUSP5/MAP3K11 in pterygium
Siying He1, Yifang Huang2, Shiqi Dong3
1Center for Gene Diagnosis, and Clinical Laboratory, Zhongnan Hospital of Wuhan University, Donghu Rd 169#, Wuhan, 430071, China.
Background:
Recently, it has been reported that miRNA is involved in pterygium, however the exact underlying mechanism in pterygium is unrevealed and require further investigation.
Methods:
The differential expression of miRNA in pterygium was profiled using microarray and validated with quantitative real-time polymerase chain reaction (qRT-PCR). Human conjunctival epithelial cells (HCEs) were cultured and treated with transforming growth factor β (TGF-β) and epidermal growth factor (EGF) and transfected with miR-199a-3p/5p mimic and inhibitor. Markers of epithelial-mesenchymal transition (EMT) in HCEs were detected using western blot and immunohistochemistry. Cell migration ability was determined using wound healing and transwell assay, while apoptosis was determined by flow cytometry. The target genes of miR-199a were confirmed by the dual-luciferase reporter assay.
Results:
TGF-β and EGF could induced EMT in HCEs and increase miR-199a-3p/5p but suppress target genes, DUSP5 and MAP3K11. With the occurrence of EMT, cell migration ability was enhanced, and apoptosis was impeded. Promoting miR-199a-3p/5p expression could induce EMT in HCEs without TGF-β and EGF, while suppressing miR-199a-3p/5p could inhibit EMT in TGF-β and EGF induced HCEs. In a word, TGF-β and EGF induced EMT could be regulated with miR-199a-3p/5p-DUSP5/MAP3K11 axes. The validated results in tissues showed that, compared with control conjunctival tissues, miR-199a-3p/5p were more overexpressed in pterygium, while DUSP5/MAP3K11 were lower expressed. In addition, bioinformatics analysis indicated the miR-199a-3p/5p-DUSP5/MAP3K11 was belong to MAPK signalling pathway.
Conclusions:
TGF-β and EGF induce EMT of HCEs through miR-199a-3p/5p-DUSP5/MAP3K11 axes, which explains the pathogenesis of EMT in pterygium and may provide new targets for pterygium prevention and therapy.
Insights
Transforming growth factor-beta and epidermal growth factor induce epithelial-mesenchymal transition in pterygium via the miR-199a-3p/5p-DUSP5/MAP3K11 pathway, offering new therapeutic targets.
Area of Science:
- Ophthalmology
- Molecular Biology
- Cell Biology
Background:
- MicroRNAs (miRNAs) are implicated in pterygium pathogenesis.
- The precise molecular mechanisms underlying miRNA involvement in pterygium remain unclear.
- Further investigation is needed to elucidate these mechanisms.
Purpose of the Study:
- To investigate the role of miR-199a-3p/5p in the epithelial-mesenchymal transition (EMT) of human conjunctival epithelial cells (HCEs).
- To explore the regulatory pathway involving miR-199a-3p/5p, DUSP5, and MAP3K11 in pterygium.
- To identify potential therapeutic targets for pterygium.
Main Methods:
- Differential expression profiling of miRNAs in pterygium using microarray and quantitative real-time polymerase chain reaction (qRT-PCR).
- In vitro studies involving HCEs treated with transforming growth factor-beta (TGF-β) and epidermal growth factor (EGF), and transfected with miR-199a-3p/5p mimics/inhibitors.
- Assessment of epithelial-mesenchymal transition (EMT) markers, cell migration, apoptosis, and target gene validation using western blot, immunohistochemistry, wound healing assay, transwell assay, flow cytometry, and dual-luciferase reporter assay.
Main Results:
- TGF-β and EGF induced EMT in HCEs, upregulating miR-199a-3p/5p and downregulating DUSP5 and MAP3K11.
- EMT enhanced cell migration and inhibited apoptosis.
- miR-199a-3p/5p promoted EMT, while its suppression inhibited TGF-β/EGF-induced EMT.
- The miR-199a-3p/5p-DUSP5/MAP3K11 axis regulates TGF-β/EGF-induced EMT.
- miR-199a-3p/5p was overexpressed, and DUSP5/MAP3K11 were underexpressed in pterygium tissues.
- Bioinformatics analysis linked this axis to the MAPK signaling pathway.
Conclusions:
- TGF-β and EGF induce HCEs EMT via the miR-199a-3p/5p-DUSP5/MAP3K11 axis.
- This pathway elucidates pterygium pathogenesis.
- The identified pathway offers potential targets for pterygium prevention and therapy.
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