Role of nuclear factor of activated T cells-2 in high mobility protein box-1 release in human monocytic THP-1 cells in vitro.
- Author:
Qing ZHAO
1
;
Li WANG
;
Jie HU
;
Hui LIU
Author Information
- Publication Type:Journal Article
- MeSH: Blotting, Western; Cell Line; Cell Nucleus; Cytoplasm; metabolism; Enzyme-Linked Immunosorbent Assay; HMGB1 Protein; metabolism; Humans; Lipopolysaccharides; Monocytes; metabolism; NFATC Transcription Factors; metabolism
- From: Journal of Southern Medical University 2016;36(1):8-12
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo investigate the role of nuclear factor of activated T cells -2 (NFAT2) in release of high mobility protein box-1 (HMGB1) from human monocytic THP-1 cells in vitro.
METHODSThe level of HMGB1 release from THP-1 cells in response to lipopolysaccharide (LPS) stimulation was examined by Western blotting and enzyme-linked immunosorbent assay (ELISA). The effect of LPS stimulation on NFAT2 and HMGB1 interaction in the cytoplasm was observed by immunoprecipitation assay. HMGB1 production and release was detected in cells with specific small interfering RNA (siRNA)-mediated suppression of NFAT2 expression.
RESULTSLPS stimulated HMGB1 release from THP-1 cells. As LPS stimulation prolonged, HMGB1 concentration increased in the cell culture supernatant and decreased in the cytoplasm, and the binding between NFAT2 and HMGB1 was not detected in the cell nuclei. NFAT2 suppression by the siRNA plasmid resulted in increased HMGB1 level in the cell culture supernatant.
CONCLUSIONNFAT2 can inhibit HMGB1 release from THP-1 cells in vitro.
