The Role of miR-200b-3p in Modulating TGF-ß1-induced Injury in Human Bronchial Epithelial Cells.
Transplantation
; 103(11): 2275-2286, 2019 11.
Article
in En
| MEDLINE
| ID: mdl-31283671
ABSTRACT
BACKGROUND:
Dysregulation of microRNAs (miRNAs) has been implicated in airway diseases where transforming growth factor-ß (TGF-ß)-induced epithelial-mesenchymal transition (EMT) may contribute to pathophysiology. Our study investigated the role of miRNA-200b in TGF-ß1-induced EMT in human bronchial epithelial cells.METHODS:
NanoString nCounter miRNA assay was used to profile miRNA in control versus TGF-ß1 (1, 4, and 24 h) stimulated BEAS-2B cells. Immortalized primary bronchial epithelial cell line (BEAS-2B cells), human primary bronchial epithelial cells (PBECs), and PBECs derived post-lung transplant were transfected with miR-200b-3p mimics and EMT marker expression was examined at RNA and protein level. miRNA target studies were performed and validated using computational tools and luciferase assay. In situ hybridization was done on normal lung tissue to localize miR-200b-3p in airway epithelium.RESULTS:
miR-200b-3p was downregulated post-TGF-ß1 treatment compared with control in BEAS-2B. miR-200b-3p mimic transfection before TGF-ß1 stimulation maintained epithelial marker expression and downregulated mesenchymal cell markers at RNA and protein level in BEAS-2B cells and PBECs. Furthermore, miR-200b-3p mimics reversed established TGF-ß1-induced EMT in BEAS-2B cells. miR-200b-3p targets, ZNF532, and ZEB2 were validated as direct targets using luciferase assay. miR-200b-3p mimics suppress TGF-ß1-induced EMT via inhibition of ZNF532 and ZEB2. In situ hybridization showed that miR-200b-3p is expressed in the normal lung epithelium. Additionally, miR-200b-3p mimics inhibit EMT in the presence of TGF-ß1 in PBECs derived from lung allograft.CONCLUSIONS:
We provide proof of concept that miR-200b-3p protects airway epithelial cells from EMT. Manipulating miR-200b-3p expression may represent a novel therapeutic modulator in airway pathophysiology.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Bronchi
/
MicroRNAs
/
Epithelial Cells
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Transforming Growth Factor beta1
/
Epithelial-Mesenchymal Transition
Limits:
Humans
Language:
En
Journal:
Transplantation
Year:
2019
Document type:
Article
Affiliation country:
United kingdom