Abstract
Bronchial epithelial cells represent an invaluable tool to elucidate molecular signaling regulation in cystic fibrosis (CF). CF is a lethal genetic condition characterized by chronic inflammation in which bronchial epithelial cells play a pivotal role. Here we describe their use in analysis of microRNA (miRNA) and their target genes following a two-step RT-PCR miRNA profiling method in bronchial cell specimens from CF and control individuals where 667 human miRNA were examined. We also describe an approach to experimental modulation of these miRNA in vitro.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsReferences
Oglesby IK, McElvaney NG, Greene CM (2010) MicroRNAs in inflammatory lung disease--master regulators or target practice? Respir Res 11:148
Applied Biosystems (2010). Applied Biosystems TaqMan® Low Density Array. Available via Applied Biosystems https://www3.appliedbiosystems.com/cms/groups/mcb_support/documents/generaldocuments/cms_042326.pdf. Accessed 14 Nov 2015
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 25:402–408
Oglesby IK, Bray IM, Chotirmall SH et al (2010) miR-126 is downregulated in cystic fibrosis airway epithelial cells and regulates TOM1 expression. J Immunol 184:1702–1709
Gruenert DC, Willems M, Cassiman JJ, Frizzell RA (2004) Established cell lines used in cystic fibrosis research. J Cyst Fibros 3(Suppl 2):191–196
Gene Expression Omnibus. NCBI. www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE19431
Primer3 Database (v4.0.0). http://frodo.wi.mit.edu. Accessed 15 Nov 2015
Untergrasser A, Cutcutache I, Koressaar T et al (2012) Primer3—new capabilities and interfaces. Nucleic Acids Res 40(15), e115
NCBI. Primer-BLAST. Available via http://www.ncbi.nlm.nih.gov/tools/primer-blast. Accessed 15 Nov 2015
NCBI. Basic Local Alignment Search Tool. Available via http://www.ncbi.nlm.nih.gov/blast. Accessed 15 Nov 2015
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2017 Springer Science+Business Media New York
About this protocol
Cite this protocol
Oglesby, I.K., McKiernan, P.J. (2017). MiRNA Expression in Cystic Fibrosis Bronchial Epithelial Cells. In: Rani, S. (eds) MicroRNA Profiling. Methods in Molecular Biology, vol 1509. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6524-3_7
Download citation
DOI: https://doi.org/10.1007/978-1-4939-6524-3_7
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-6522-9
Online ISBN: 978-1-4939-6524-3
eBook Packages: Springer Protocols