title: Comparison of miRNA profiling during airway epithelial repair in undifferentiated and differentiated cells in vitro.
abstract: Respiratory epithelium is a highly integrated structure that efficiently protects lungs from extrinsic irritants thanks to rapid repair of the wound. The repair is a complex process that requires coordinated expression of networks of genes. Plausible regulators of this process are microRNAs. We investigated whether global miRNA silencing influences the epithelial repair, and whether changes in miRNA expression profile during repair are similar between two bronchial epithelial cell cultures: differentiated and undifferentiated cells. Two bronchial cell types were used:16HBE14o- and NHBE. Transfection was performed with siRNAs against Drosha and Dicer. For miRNA profiling, non-transfected cells were cultured until confluent and harvested for RNA isolation at baseline (cells before wounding) and at different time post-wounding (8, 16, 24, and 48 h). MicroRNA expression profiling was performed using TaqMan Array Human MicroRNA Card A. Target prediction was done in miRNA body map, and pathway analysis using DAVID. Cells with downregulated Drosha and Dicer demonstrated a significantly delayed wound repair in comparison to control in both cell lines. MiRNA expression profiling revealed that ten miRNAs exhibited significant changes over time after cell injury. These genes showed a similar expression pattern in both cell lines. The predicted targets of these miRNAs were then clustered by pathway analysis into six biological groups related to wound repair. Silencing of global miRNA expression confirmed that miRNAs are crucial for airway epithelial repair. Moreover, epithelial cells of two different origins demonstrated some similarities in miRNA expression pattern during wound repair, independent of differentiation state.
authors: Laboratory of Molecular and Cell Biology, Department of Pediatric Pulmonology, Allergy and Clinical Immunology, Poznan University of Medical Sciences, 27/33 Szpitalna St., 60-572, Poznan, Poland.
authors: Wojciech
authors: W
authors: Langwinski
authors: Laboratory of Molecular and Cell Biology, Department of Pediatric Pulmonology, Allergy and Clinical Immunology, Poznan University of Medical Sciences, 27/33 Szpitalna St., 60-572, Poznan, Poland.
authors: Beata
authors: B
authors: Narozna
authors: Clinical and Experimental Sciences, Faculty of Medicine, University of Southampton, Southampton, UK.
authors: Peter M
authors: PM
authors: Lackie
authors: Clinical and Experimental Sciences, Faculty of Medicine, University of Southampton, Southampton, UK.|Human Development and Health, Faculty of Medicine, University of Southampton, Southampton, UK.
authors: John W
authors: JW
authors: Holloway
authors: Laboratory of Molecular and Cell Biology, Department of Pediatric Pulmonology, Allergy and Clinical Immunology, Poznan University of Medical Sciences, 27/33 Szpitalna St., 60-572, Poznan, Poland. alszczep@ump.edu.pl.
authors: Aleksandra
authors: A
authors: Szczepankiewicz
mesh_terms: D002454:Cell Differentiation; D002460:Cell Line; D053487:DEAD-box RNA Helicases / Q000235:genetics; D015536:Down-Regulation; D004847:Epithelial Cells / Q000166:cytology*; D020869:Gene Expression Profiling; D020868:Gene Silencing; D006801:Humans; D035683:MicroRNAs / Q000235:genetics*; D043244:Ribonuclease III / Q000235:genetics; D014162:Transfection; D014945:Wound Healing / Q000235:genetics*
chemical_list: D035683:MicroRNAs; C408087:DICER1 protein, human; C517918:DROSHA protein, human; D043244:Ribonuclease III; D053487:DEAD-box RNA Helicases
keywords: 16HBE14o- cells; Airway epithelium; NHBE cells; Wound repair; miRNA