GATA3 siRNA inhibits the binding of NFAT1 to interleukin-13 promoter in human T cells.
- Author:
Xin YAO
1
;
Yan YANG
;
Hai-yan HE
;
Min WANG
;
Kai-sheng YIN
;
Mao HUANG
Author Information
- Publication Type:Journal Article
- MeSH: Cells, Cultured; GATA3 Transcription Factor; antagonists & inhibitors; genetics; Humans; Interleukin-13; genetics; NFATC Transcription Factors; metabolism; Promoter Regions, Genetic; RNA, Small Interfering; genetics; T-Lymphocytes; metabolism; Transfection
- From: Chinese Medical Journal 2010;123(6):739-744
- CountryChina
- Language:English
-
Abstract:
BACKGROUNDInterleukin-13 (IL-13) is recognized to be a key modulator in the pathogenesis of Th2-induced allergic inflammation. Transcription factors GATA3 and NFAT1 have been both implicated in the regulation of Th2 cytokines. We previously demonstrated the GATA3-NFAT1 association during human T cell activation. However, the function of the GATA3-NFAT1 complex in Th2 cytokines regulation is still unknown. Small interference RNA (siRNA) was constructed to knock down GATA3 expression in Hut-78 cells to investigate the possible role of GATA3-NFAT1 complex in IL-13 transcription.
METHODSCells were stimulated with anti-CD3 plus anti-CD28 antibodies to mimic in vivo antigen-mediated co-stimulation; the expression of IL-13 mRNA was determined by real-time PCR; chromation immunoprecipitation (CHIP) assay was employed to investigate the NFAT1 binding to IL-13 promoter.
RESULTSGATA3 siRNA suppressed the expression of GATA3 both in mRNA and protein levels in Hut-78 cells. The binding of NFAT1 to IL-13 promoter was inhibited by GATA3 siRNA in activated T cells, which was followed by the reduction of IL-13 transcription.
CONCLUSIONGATA3-NFAT1 complex may play an important role in the regulation of IL-13 transcription in human T cells.
