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-6 and 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 DC2.4 cell line stably overexpressing the target gene, and detected the TNF-α secretion after transfecting miR-144 mimics. Results The miR-144 expression was significantly down-related in DCs stimulated with LPS(P<0.01). After the miR-144 mimics were transfected into DCs,expressions of TNF-α,IL-1β,IL-6 and 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 DC2.4 cells stably overexpressing RANKL resulted in no change of TNF-α mRNA expression. Conclusion miR-144 can regulate DCs cytokine secretion by targeting RANKL.