Effects of Bcl-2 and Bcl-XL protein levels on chemoresistance of hepatoblastoma HepG2 cell line

Biochem Cell Biol. 2000;78(2):119-26.

Abstract

The ratio between apoptotic promoters and repressors in the Bcl-2 family determines the chemosensitivity of cells to apoptotic stimuli. This study examines the chemoresistance of a transfected human hepatoblastoma HepG2 cell-line during Taxol and Doxorubicin application. Sense bcl-2, and anti-sense bcl-XL gene fragments were separately inserted into HepG2 cells via stable transfection. The expression profile of the Bcl-2 family proteins was determined by Western blot analysis. Chemosensitivity of the transfected cells was measured by Trypan blue exclusion assay and XTT reduction assay during drug application. In the absence of Bax protein, HepG2 cells with elevated Bcl-2 protein levels did not exhibit any significant increase in chemosensitivity towards the drugs. Transfected cells with reduced Bcl-XL levels became more sensitive to the drugs, and a significant difference in IC50 values was observed. The chemosensitivity of HepG2 cells to Taxol and Doxorubicin was not affected by Bcl-2 levels, while reduction of Bcl-XL levels rendered the cells more sensitive to the drugs. This suggests that the Bcl-2 protein alone could not protect HepG2 cells from drug-induced apoptosis, and that the Bcl-XL protein may be a target for gene therapy in hepatoblastoma treatment.

MeSH terms

  • Antineoplastic Agents / pharmacology
  • Antineoplastic Agents, Phytogenic / pharmacology
  • Apoptosis
  • Blotting, Western
  • Cell Line
  • Cell Survival / drug effects
  • Dose-Response Relationship, Drug
  • Doxorubicin / pharmacology
  • Drug Resistance, Neoplasm*
  • Genes, bcl-2 / genetics
  • Hepatoblastoma / drug therapy
  • Humans
  • Inhibitory Concentration 50
  • Liver Neoplasms / drug therapy
  • Paclitaxel / pharmacology
  • Plasmids / metabolism
  • Promoter Regions, Genetic
  • Proto-Oncogene Proteins c-bcl-2 / genetics
  • Proto-Oncogene Proteins c-bcl-2 / metabolism*
  • Time Factors
  • Transfection
  • Tumor Cells, Cultured
  • bcl-X Protein

Substances

  • Antineoplastic Agents
  • Antineoplastic Agents, Phytogenic
  • BCL2L1 protein, human
  • Proto-Oncogene Proteins c-bcl-2
  • bcl-X Protein
  • Doxorubicin
  • Paclitaxel