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Using multiple online databases to help identify micro RNA s regulating the airway epithelial cell response to a virus‐like stimulus
Author(s) -
Herbert Cristan,
Sebesfi Michelle,
Zeng QingXiang,
Oliver Brian G,
Foster Paul S,
Kumar Rakesh K
Publication year - 2015
Publication title -
respirology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.857
H-Index - 85
eISSN - 1440-1843
pISSN - 1323-7799
DOI - 10.1111/resp.12606
Subject(s) - rna , gene expression , gene , microrna , virus , stimulus (psychology) , medicine , real time polymerase chain reaction , database , cell , biology , microbiology and biotechnology , immunology , genetics , computer science , psychology , psychotherapist
Abstract Background and objective Exacerbations of allergic asthma are often triggered by respiratory viral infections. We have previously shown that in a T ‐helper type 2 ( T h2)‐biased cytokine environment, mouse and human airway epithelial cells ( AEC ) exhibit increased expression of pro‐inflammatory and anti‐viral genes in response to synthetic double‐stranded ribonucleic acid (ds RNA ), a virus‐like stimulus. This implies coordinated regulation of gene expression, suggesting possible involvement of micro RNA . To investigate this, we developed a novel approach to identifying candidate micro RNA using online databases, then confirmed their expression by quantitative real‐time polymerase chain reaction ( qRT ‐ PCR ). Methods Using a list of genes of interest, defined on the basis of the previous study as being up‐regulated in a T h2 environment, we searched mouse and human micro RNA databases for possible regulatory micro RNA , and selected 10 candidates that were conserved across species or predicted by more than one human database. Expression of these micro RNA was tested by qRT ‐ PCR , in primary human AEC pre‐treated with T h2 cytokines and exposed to ds RNA . Results Expression of hsa‐mi R ‐139‐5p, mi R ‐423‐5p and mi R ‐542‐3p was significantly decreased in T h2 pre‐treated AEC , and mi R ‐135a‐5p exhibited a trend towards decreased expression. Further database searches confirmed that these micro RNA regulated additional pro‐inflammatory and anti‐viral response genes for which expression had previously been shown to be up‐regulated, confirming the validity of this approach. Conclusions Our study demonstrates the value of using multiple online databases to identify candidate regulatory micro RNA and provides the first evidence that in an allergic environment, micro RNA may be important in altering the pro‐inflammatory and anti‐viral responses of human AEC during exacerbations of asthma.