Antitumor activity of cationic liposome-mediated Bax gene transfer in osteosarcoma cells: induction of apoptosis and caspase-independent cell death

Int J Oncol. 2005 Aug;27(2):433-8.

Abstract

The purpose of this study was to evaluate the anti-tumor effects of osteosarcoma (HOSM-1) cells via transfer of the Bax gene using a cationic liposome. We evaluated the levels of Bax, Bcl-xL, Bcl-2 and cytochrome c expression by Western blot analysis, and caspase-9 and -3 activities were determined in a colorimetric assay. Apoptosis was detected using a TUNEL assay, and cell growth inhibition was determined in an MTT assay. Following Bax gene transfer, release of cytochrome c to the cytosol was detected, the activities of caspase-9 and -3 increased, and TUNEL-positive cells (37.5%) were detected. Cell survival rate was 50.8% under these conditions. Induction of apoptosis was inhibited by a caspase inhibitor (zVAD-fmk), but only a slight increase in cell survival rate occurred. Hence, since not only apoptosis but also caspase-independent cell death is induced in HOSM-1 cells, we anticipate that Bax gene therapy with cationic liposomes will be useful for osteosarcoma.

Publication types

  • Comparative Study

MeSH terms

  • Amino Acid Chloromethyl Ketones / pharmacology
  • Apoptosis / drug effects
  • Apoptosis / genetics
  • Blotting, Western
  • Caspase 3
  • Caspase 9
  • Caspase Inhibitors
  • Caspases / metabolism
  • Cations / chemistry
  • Cell Line, Tumor
  • Cell Survival / drug effects
  • Cell Survival / genetics
  • Humans
  • In Situ Nick-End Labeling
  • Liposomes / chemistry
  • Osteosarcoma / enzymology
  • Osteosarcoma / genetics
  • Osteosarcoma / pathology
  • Transfection / methods*

Substances

  • Amino Acid Chloromethyl Ketones
  • Caspase Inhibitors
  • Cations
  • Liposomes
  • benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone
  • CASP3 protein, human
  • CASP9 protein, human
  • Caspase 3
  • Caspase 9
  • Caspases