ABSTRACT: Real-time quantitative PCR analysis of microRNA expression in Primary human Bronchial Epithelial cells (PBECs) from healthy or asthmatic subjects
Project description:Bronchial asthma is one of the most common respiratory diseases. Reversible airflow limitation, persistent airway inflammation and airway hyperresponsiveness, and air remodeling is its main characterization. The morbidity and mortality rates of asthma increase due to air pollution. MicroRNAs are small molecules that regulate gene expression playing a part in inflammatory diseases. The designed current study aims to compare and screen circulating microRNAs targeting LIGHT as diagnostic biomarkers during asthma attack through microRNA microarray and Real Time PCR assay. Methods: Serum from 40 patients with asthma attack and 20 normal subjects was collected to detect circulating microRNAs expression profile through miRNA microarray. It is revealed that the significant differences of microRNAs level between asthmatic patients and normal controls. Target gene predictive analysis of differentially expressing microRNAs was performed to screen the miRNAs targeting LIGHT gene basing on the TargetScan bioinformatics software. Real Time PCR was conducted to further verify the realiability of the miRNA microassay and the screened specific miRNAs targeting LIGHT. Results: Asthmatic patients had a significant upper expression of miR-512-3p, miR-513b-5p, miR-5691 and lower level of miR-107, miR-140-5p, miR-17-5p compared with healthy subjects. The level of miR-140-5p and miR-107 in plasma of asthmatic patients were tightly correlated with their eosinophilic inflammation. The plasma miR-140-5p and miR-107 were needed to perform further investigation to evaluate the role as biomarkers in diagnosis of asthma.
Project description:Airway inflammation is the hallmark of asthma and suggests a dysregulation of homeostatic mechanisms. MicroRNAs (miRNAs) are key regulators of gene expression, necessary for the proper function of cellular processes. We used miRNA microarrays to compare the profiles of human bronchial epithelial cells from healthy and asthmatic donors Human bronchial epithelial cells were isolated from healthy and asthmatic donors. RNA was extracted and miRNA expression was analyzed on Affymetrix miRNA microarrays. We sought to utilize miRNA expression as a tool for understanding underlying biological differences in BECs from healthy and asthmatic donors.
