Regulatory effect of miR-144 on cytokine secretion in dendritic cells by targeting receptor activator of NF-κB ligand
10.3724/SP.J.1008.2014.01053
- Author:
Meng GUO
1
Author Information
1. Department of Organ Transplantation, Changzheng Hospital, Second Military Medical University
- Publication Type:Journal Article
- Keywords:
Cytokines;
Dendritic cells;
miRNA-144;
Receptor activator ofNF-κB ligand
- From:
Academic Journal of Second Military Medical University
2014;35(10):1053-1059
- CountryChina
- Language:Chinese
-
Abstract:
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-1Jβ, 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 veritication. 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-1/3, IL-6 and JL-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-kB 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.
