Inhibition of tissue factor by siRNA enhances doxorubicin-induced apoptosis in human neuroblastoma.
- Author:
Jun FANG
1
;
Hao TANG
;
Ling-Hui XIA
;
Mu-Xiang ZHOU
;
Yan CHEN
;
Wen-Ning WEI
;
Yu HU
;
Shan-Jun SONG
Author Information
- Publication Type:Journal Article
- MeSH: Apoptosis; drug effects; genetics; Caspase 3; metabolism; Cell Line, Tumor; Doxorubicin; pharmacology; Genetic Vectors; Humans; Neuroblastoma; metabolism; pathology; Poly(ADP-ribose) Polymerases; metabolism; RNA Interference; RNA, Small Interfering; genetics; Thromboplastin; genetics; metabolism; Transfection
- From: Chinese Journal of Hematology 2007;28(9):594-597
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo investigate the regulation of tissue factor (TF) on doxorubicin-induced apoptosis in human neuroblastoma.
METHODThe expression of TF was examined by Western blotting. TF siRNA-pSUPER plasmid was constructed by inserting a specific 19-nt silencing sequence targeting TF gene into pSUPER vector. Transfection of TF siRNA-pSUPER was performed using lipofectamine 2000. The activation of caspase-3 and PARP induced by doxorubicin was tested by Western blotting. The apoptotic cells were stained by Hochest 33342 and counted under fluorescence inverted microscope.
RESULTS(1) Human neuroblastoma cell line SK-N-MC expressed high level of TF. (2) Downregulation of TF expression was achieved by transfection of TF siRNA-pSUPER into SK-N-MC cells in a dose-dependent manner. (3) Cleavage of caspase-3 and PARP was increased in transfected SK-N-MC cell with down-regulation of TF. (4) TF siRNA treatment at 1 microg/ml for 8 h significantly increased apoptotic cell number in transfected SK-N-MC cells compared to that in non-transfected cells (P < 0.05) while exposing to 1 microg/ml doxorubicin for 8 h.
CONCLUSIONSDownregulation of TF expression by specific siRNA vector could increase the cytotoxicity of doxorubicin and enhance doxorubicin-induced apoptosis in human neuroblastoma cells.
