A functional polymorphism at the miR­491­5p binding site in the 3'­untranslated region of the MMP­9 gene increases the risk of developing ventilator­associated pneumonia.

ABSTRACT

Matrix metalloproteinase (MMP)­9 is associated with the severity of ventilator­associated pneumonia (VAP), while an rs1056629 SNP located in the 3'­untranslated region (UTR) of MMP­9 affects the microRNA (miRNA/miR)­491­mediated regulation of MMP­9 expression. In the present study, the effect of rs1056629 on the development of VAP in patients with chronic obstructive pulmonary disease (COPD) was investigated. Patients with COPD were enrolled in the study and their genotypes of rs1056629 (CC, CA or AA) were determined. ELISA was used to analyze the levels of TNF­α and IL­6 in the monocytes of patients with COPD carrying differential genotypes of rs1056629. Reverse transcription­quantitative PCR was carried out to evaluate the expression of miR­491 and MMP­9 mRNA in the different groups of patients with COPD. Luciferase assay was used to confirm the inhibitory role of miR­491 in MMP­9 expression. Western blot analysis was carried out to assess the expression of MMP­9 protein in A549 and H1299 cells transfected with miR­491 mimics. The risk and severity of VAP were significantly elevated in patients with COPD carrying the CC and AC genotypes of rs1056629. Although there was no difference in the expression of miR­491 in patients carrying different genotypes of rs1056629, the expression levels of TNF­α, IL­6 and MMP­9 were increased in patients with COPD carrying the CC and AC genotypes of rs1056629. The results of luciferase assay revealed that miR­491 inhibited the expression of MMP­9 through direct binding to the 3'UTR of MMP­9. Transfection of miR­491 mimics into A549 and H1299 cells markedly suppressed the expression of MMP­9 in a concentration­dependent manner. On the whole, the findings of the present study confirm that the CC and AC genotypes of rs1056629 increase the risk of developing VAP in patients with COPD by increasing the expression of MMP­9.