MicroRNA‐16 regulates ENaC expression in alveolar epithelial cells

Abstract

ObjectiveMicroRNAs (miRNAs) have emerged as a novel class of gene regulators which play a critical role in complex diseases like acute lung injury (ALI). Our miRNA profiling studies in animal models of ALI revealed that miR‐146a, mir‐16 and miR‐155 were dysregulated. Mir16 targets Serotonin transporter (SERT) which is involved in serotonin uptake. Elevated levels of serotonin in ALI are known to regulate epithelial sodium channel (ENac) expression. The aim of this study is to investigate the functional effects of mir16 on SERT and ENaC in human alveolar epithelial cells (A549) in vitro.MethodsA549 cells were commercially obtained and cultured according to the manufacturer instructions. Cells were transfected with plasmid containing miR16 expressing or control vector by Lipofectamine. q PCR and immunoblot analysis were performed to assess the effects of miR‐16 on SERT, ENaC and other downstream signalling molecules.ResultsA549 cells transfected with miR16 decreased the expression levels of SERT when compared to controls. Dose dependant regulation of SERT was observed in untransfected A549 cells treated with serotonin. Increased expression of ENaC β was observed in miR16 transfected cells compared to controls.ConclusionmiR16 over expression altered SERT and ENaC expression in human alveolar epithelial cells.Research supported by:RO1HL105932(NIH) and 09SDG2260957(AHA)