Effect of siRNA-mediated inhibition of nuclear transcription factor-kappa B on apoptosis of hepatocarcinoma cells.
- Author:
Yi-lang WANG
1
;
Deng-fu YAO
;
Wei WU
;
Wen-li SAI
;
Li-wei QIU
;
Jun-ling YANG
;
Jian-wei ZHU
Author Information
- Publication Type:Journal Article
- MeSH: Apoptosis; drug effects; Carcinoma, Hepatocellular; metabolism; pathology; Gene Expression Regulation; Hep G2 Cells; Humans; Liver Neoplasms; metabolism; pathology; NF-kappa B; antagonists & inhibitors; metabolism; RNA, Small Interfering; pharmacology
- From: Chinese Journal of Hepatology 2010;18(8):609-613
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo investigate the effect of siRNA-mediated inhibition of NF-κB on apoptosis of hepatocarcinoma cells.
METHODSSpecific small interfering RNA Targeting NF-κB gene was synthesized and transfected into HepG2 cells by liposomes. Nested RT-PCR and quantitative RT-PCR were used to detect the mRNA expression of NF-κB. Immunohistochemistry, enzyme-linked immunosorbent assay and Western blot were performed to examine the protein expression of NF-κB. Annexin V-FITC was used to test cell apoptosis.
RESULTSThe expression of NF-κB in HepG2 cells (1.13+/-0.03) was significantly higher (t=27.02, P<0.05) than that in normal hepatocytes (0.29+/-0.07). The down-regulation of NF-κB expression was depended on the dosage of siRNA and the time after transfection. 72 h after siRNA transfection, NF-κB expression reduced by 93% and 62% at the mRNA and protein levels, respectively. The apoptosis of HepG2 cells increased by 85% with NF-κB inhibition.
CONCLUSIONSNF-κB is abnormally active in HepG2 cells and NF-κB inhibition mediated by siRNA promotes HepG2 cells apoptosis. It suggested that NF-κB could be a potential target for HCC prevention gene therapy.
