Comparison of caspase genes for the induction of apoptosis following gene delivery

Mol Biotechnol. 2009 Mar;41(3):236-46. doi: 10.1007/s12033-008-9133-9. Epub 2008 Dec 9.

Abstract

The polycation poly(ethylenimine) (PEI) was used to deliver the plasmids coding for various combinations of caspases to Cox-2 overexpressing cancer cell lines. It was found that the expression of the delivered genes, controlled by the Cox-2 promoter, correlated with the expression of the endogenous Cox-2 gene in each cell line in a relatively linear manner. Among the various caspase combination regimens, the combination of caspase 3 plus caspase 9 proved to be the most effective because of an apparent synergy between the two gene products, and produced phosphatidylserine flipping in addition to fragmentation of genomic DNA. Caspase 1 appeared to work independently of either caspases 3 or 9, as no synergistic effect was observed. Transfections with genes coding for granzyme B and caspase 8 yielded a lesser amount of cell death. The delivery of a combination of caspase genes could be readily moved to in vivo research of bladder and colon cancer treatments, and holds great applicability to a wide array of additional tumor types.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Analysis of Variance
  • Animals
  • Apoptosis*
  • Carcinoma, Transitional Cell / enzymology
  • Caspases / genetics*
  • Caspases / metabolism
  • Cell Line, Tumor
  • Cell Survival / genetics
  • Cloning, Molecular
  • Colonic Neoplasms / enzymology
  • Cyclooxygenase 2 / genetics
  • Cyclooxygenase 2 / metabolism*
  • Fibroblasts / enzymology
  • Fluorescein-5-isothiocyanate
  • Granzymes / genetics
  • Granzymes / metabolism
  • In Situ Nick-End Labeling
  • Mice
  • Polyethyleneimine / adverse effects
  • RNA, Messenger / metabolism
  • Transfection*
  • Urinary Bladder Neoplasms / enzymology

Substances

  • RNA, Messenger
  • Polyethyleneimine
  • Ptgs2 protein, mouse
  • Cyclooxygenase 2
  • Granzymes
  • Gzmb protein, mouse
  • Caspases
  • Fluorescein-5-isothiocyanate