Comparison of miRNA profiling during airway epithelial repair in undifferentiated and differentiated cells in vitro
Comparison of miRNA profiling during airway epithelial repair in undifferentiated and differentiated cells in vitro
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
205-212
Langwinski, Wojciech
f9de82f2-03f0-4b85-b70e-5d2433bed346
Narozna, Beata
1c6178db-d6f1-4f24-873e-38a8c47357d8
Lackie, Peter M.
4afbbe1a-22a6-4ceb-8cad-f3696dc43a7a
Holloway, John W.
4bbd77e6-c095-445d-a36b-a50a72f6fe1a
Szczepankiewicz, Aleksandra
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May 2017
Langwinski, Wojciech
f9de82f2-03f0-4b85-b70e-5d2433bed346
Narozna, Beata
1c6178db-d6f1-4f24-873e-38a8c47357d8
Lackie, Peter M.
4afbbe1a-22a6-4ceb-8cad-f3696dc43a7a
Holloway, John W.
4bbd77e6-c095-445d-a36b-a50a72f6fe1a
Szczepankiewicz, Aleksandra
54b47a55-9a85-44c8-b4b9-dea4a8d9fe73
Langwinski, Wojciech, Narozna, Beata, Lackie, Peter M., Holloway, John W. and Szczepankiewicz, Aleksandra
(2017)
Comparison of miRNA profiling during airway epithelial repair in undifferentiated and differentiated cells in vitro.
Journal of Applied Genetics, 58 (2), .
(doi:10.1007/s13353-016-0370-5).
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
Text
Manuscript_JOAG_revised_final
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More information
Accepted/In Press date: 11 October 2016
e-pub ahead of print date: 28 October 2016
Published date: May 2017
Organisations:
Human Development & Health, Clinical & Experimental Sciences
Identifiers
Local EPrints ID: 402329
URI: http://eprints.soton.ac.uk/id/eprint/402329
ISSN: 1234-1983
PURE UUID: cea360ee-b0cd-40c5-bfa9-8545af359358
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Date deposited: 04 Nov 2016 15:20
Last modified: 10 Dec 2019 06:21
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Contributors
Author:
Wojciech Langwinski
Author:
Beata Narozna
Author:
Aleksandra Szczepankiewicz
University divisions
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