title: GATA3 siRNA inhibits the binding of NFAT1 to interleukin-13 promoter in human T cells.
abstract: BACKGROUND
  Interleukin-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.
  
  
  METHODS
  Cells 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.
  
  
  RESULTS
  GATA3 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.
  
  
  CONCLUSIONS
  GATA3-NFAT1 complex may play an important role in the regulation of IL-13 transcription in human T cells.
authors.affiliation: Department of Respiratory Disease, First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu 210029, China.
authors.forename: Xin
authors.initials: X
authors.lastname: Yao
authors.forename: Yan
authors.initials: Y
authors.lastname: Yang
authors.forename: Hai-yan
authors.initials: HY
authors.lastname: He
authors.forename: Min
authors.initials: M
authors.lastname: Wang
authors.forename: Kai-sheng
authors.initials: KS
authors.lastname: Yin
authors.forename: Mao
authors.initials: M
authors.lastname: Huang
mesh_terms: D002478:Cells, Cultured; D050990:GATA3 Transcription Factor / Q000037:antagonists & inhibitors* / Q000235:genetics; D006801:Humans; D018793:Interleukin-13 / Q000235:genetics*; D050778:NFATC Transcription Factors / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D034741:RNA, Small Interfering / Q000235:genetics*; D013601:T-Lymphocytes / Q000378:metabolism*; D014162:Transfection
chemical_list: D050990:GATA3 Transcription Factor; C494749:GATA3 protein, human; D018793:Interleukin-13; D050778:NFATC Transcription Factors; C494006:NFATC2 protein, human; D034741:RNA, Small Interfering