title: [Effect of siRNA-mediated inhibition of nuclear transcription factor-kappa B on apoptosis of hepatocarcinoma cells].
abstract: OBJECTIVE
  To investigate the effect of siRNA-mediated inhibition of NF-κB on apoptosis of hepatocarcinoma cells.
  
  
  METHODS
  Specific 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.
  
  
  RESULTS
  The 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.
  
  
  CONCLUSIONS
  NF-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.
authors.affiliation: Research Center of Clinical Medicine, Affiliated Hospital of Nantong University, Nantong 226001, China.
authors.forename: Yi-lang
authors.initials: YL
authors.lastname: Wang
authors.forename: Deng-fu
authors.initials: DF
authors.lastname: Yao
authors.forename: Wei
authors.initials: W
authors.lastname: Wu
authors.forename: Wen-li
authors.initials: WL
authors.lastname: Sai
authors.forename: Li-wei
authors.initials: LW
authors.lastname: Qiu
authors.forename: Jun-ling
authors.initials: JL
authors.lastname: Yang
authors.forename: Jian-wei
authors.initials: JW
authors.lastname: Zhu
mesh_terms: D017209:Apoptosis / Q000187:drug effects*; D006528:Carcinoma, Hepatocellular / Q000378:metabolism* / Q000473:pathology; D005786:Gene Expression Regulation; D056945:Hep G2 Cells; D006801:Humans; D008113:Liver Neoplasms / Q000378:metabolism* / Q000473:pathology; D016328:NF-kappa B / Q000037:antagonists & inhibitors / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000494:pharmacology*
chemical_list: D016328:NF-kappa B; D034741:RNA, Small Interfering