siRNA-mediated beta-catenin knockdown in human hepatoma cells results in decreased growth and survival

Neoplasia. 2007 Nov;9(11):951-9. doi: 10.1593/neo.07469.


beta-Catenin, the chief oncogenic component of the canonical Wnt pathway, is known to be involved in a variety of cancers, including hepatocellular carcinoma (HCC). Although the mechanism of beta-catenin activation in HCC is multifactorial, it is indisputably implicated at various stages of hepatocarcinogenesis, making it an attractive therapeutic target. Here we investigate the effect of small interfering RNA-mediated beta-catenin knockdown on the growth and survival of human hepatoma cell lines with (HepG2) and without (Hep3B) beta-catenin mutations. Transfection of HepG2 and Hep3B cells with human beta-catenin (CTNNB1) small interfering RNA resulted in a significant beta-catenin decrease, as confirmed by Western blot analyses and immunofluorescence, also leading to decreased expression of known target genes such as cyclin D1 and glutamine synthetase. The decrease in beta-catenin activity was confirmed by TOPflash reporter luciferase assay. The functional impact of diminished beta-catenin was exhibited as temporal decrease in tumor cell viability by the MTT assay. A concomitant decrease in tumor cell proliferation was also evident with [(3)H]thymidine incorporation and verified with soft agar assays. Thus, beta-catenin is essential for the survival and growth of hepatoma cells independent of mutations in the beta-catenin gene and provide a proof of principle for the significance of the therapeutic inhibition of beta-catenin in HCC.

Keywords: Liver cancer; Wnt/β-catenin; development; regeneration; treatment.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Carcinoma, Hepatocellular / pathology
  • Carcinoma, Hepatocellular / therapy*
  • Cell Line, Tumor
  • Cell Proliferation
  • Cell Survival
  • Cyclin D1 / analysis
  • Fluorescent Antibody Technique
  • Glutamate-Ammonia Ligase / analysis
  • Glycogen Synthase Kinase 3 / physiology
  • Glycogen Synthase Kinase 3 beta
  • Humans
  • Liver Neoplasms / pathology
  • Liver Neoplasms / therapy*
  • Mutation
  • RNA, Small Interfering / therapeutic use*
  • beta Catenin / antagonists & inhibitors*
  • beta Catenin / genetics


  • RNA, Small Interfering
  • beta Catenin
  • Cyclin D1
  • Glycogen Synthase Kinase 3 beta
  • Glycogen Synthase Kinase 3
  • Glutamate-Ammonia Ligase