Identification of miR-328-3p targets during wound repair of bronchial epithelial cells

Abstract

<b>Background:</b> Wound repair of injured airway epithelium requires the phased expression of networks of genes regulated by microRNAs, a mechanism that might be disturbed in diseases characterized by aberrant repair and chronic inflammation. This research aimed to investigate which mRNAs are targeted by microRNA-328-3p, previously shown to be crucial during the repair of airway epithelium. <b>Methods:</b> We transfected the cell culture model of human bronchial epithelium with miR-328 inhibitor and performed a wound assay. Whole transcriptome human microarrays (Agilent) and GeneSpring 14.9 software were used to analyze the mRNA expression at four time points: before the injury and 4, 8, 16 hours after injury. We compared results to databases containing the predicted and validated targets of miR-328. <b>Results:</b> As a result of miR-328 inhibition, the expression levels of 3669 genes were significantly altered during the wound repair (fold change&gt;2.0, p&lt;0.05). Six genes were common for all time points, whereas others were either differentially expressed at only one time point or were shared between two or three time points, implicating their involvement at a specific stage of the epithelial wound repair. Bioinformatics analysis has also confirmed that 897 of these genes are predicted targets of miR-328. <b>Conclusion:</b> Inhibition of miR-328 during wound repair had altered gene expression of numerous genes at different time points, suggesting that they might be either direct or indirect targets of this miR and therefore are involved during the repair of the injured airway epithelium. This study was supported by the Polish National Science Centre grant no. 2017/25/N/NZ3/00332.