Regulatory effect of miR-144 on cytokine secretion in dendritic cells by targeting receptor activator of NF-κB ligand

Objective To investigate the effect of miRNA-144 on cytokine secretion by dendritic cells(DCs)during their maturation and the related mechanism.Methods miRNA chip results showed markedly down-regulated miRNA-144 during DCs maturation.The miR-144 level was also observed in DCs before and after treatment with lipopolysaccharide(LPS).The changes of related cytokines(TNF-α,IL-1β,IL-6and IL-23)and the activation of signaling pathway(NF-κB and MAPK)were detected in DCs after the transfection of miR-144.TargetScan was used to predict the target spot of miR-144 and double fluorescence reporting system was used for verification.Furthermore,we established DC2.4cell line stably overexpressing the target gene,and detected the TNF-αsecretion after transfecting miR-144 mimics.Results The miR-144 expression was significantly down-related in DCs stimulated with LPS(P0.01).After the miR-144 mimics were transfected into DCs,expressions of TNF-α,IL-1β,IL-6and IL-23 mRNA were significantly reduced(P 0.05,P 0.01),and NF-κB phosphorylation level was reduced as well.Bioinformatics analysis hinted that receptor activator of NF-κB ligand gene(RNAKL)was the potential target of miR-144,which was also validated by double fluorescence reporting system.Moreover,transfecting miR-144 mimics into DC2.4cells stably overexpressing RANKL resulted in no change of TNF-α mRNA expression.Conclusion miR-144 can regulate DCs cytokine secretion by targeting RANKL.