Exogenous wt-p53 protein is active in transformed cells but not in their non-transformed counterparts: implications for cancer gene therapy without tumor targeting

J Gene Med. 2000 Jan-Feb;2(1):11-21. doi: 10.1002/(SICI)1521-2254(200001/02)2:1<11::AID-JGM81>3.0.CO;2-K.

Abstract

Background: Expression of exogenous wild-type p53 (wt-p53) protein in tumor cells can suppress the transformed phenotype whereas it does not apparently induce detrimental effects in non-transformed cells. This observation may provide a molecular basis for p53-mediated gene therapy of p53-sensitive cancers without the need for tumor targeting.

Methods: To understand the molecular mechanisms responsible for this different behavior in tumor versus normal cells, biochemical and functional analyses of exogenous wt-p53 protein were performed on non-transformed C2C12 myoblasts and their transformed counterparts, the C2-ras cells.

Results: The exogenous wt-p53 protein, which induced persistent growth arrest only in transformed C2-ras cells, was shown to be significantly more stable in transformed than in non-transformed cells. This different stability was due to different p53 proteolytic degradation. Moreover, constitutively, exogenous wt-p53 protein was found to be transcriptionally active only in C2-ras cells but it could also be activated in C2C12 cells by genotoxic damage.

Conclusions: Non-transformed C2C12 cells present regulatory system(s) which control the expression and the activity of exogenously expressed wt-p53 protein probably through degradation and maintenance in a latent form. This regulatory system is lost/inactivated upon transformation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenoviridae / genetics
  • Animals
  • Blotting, Western
  • Cell Division
  • Cell Line
  • Cell Line, Transformed
  • Cell Transformation, Neoplastic / genetics*
  • Cell Transformation, Neoplastic / metabolism
  • Chloramphenicol O-Acetyltransferase / genetics
  • Chloramphenicol O-Acetyltransferase / metabolism
  • Cysteine Endopeptidases / pharmacology
  • DNA Damage / physiology
  • DNA, Recombinant / genetics
  • Doxorubicin / pharmacology
  • Gene Expression Regulation, Neoplastic*
  • Gene Transfer Techniques
  • Genes, Reporter / genetics
  • Genes, ras / genetics
  • Genetic Vectors / genetics
  • Leupeptins / pharmacology
  • Mice
  • Mice, Inbred C3H
  • Multienzyme Complexes / pharmacology
  • Muscles / cytology
  • Neoplasm Proteins / genetics
  • Neoplasm Proteins / metabolism
  • Nuclear Proteins*
  • Proteasome Endopeptidase Complex
  • Proto-Oncogene Proteins / genetics
  • Proto-Oncogene Proteins / metabolism
  • Proto-Oncogene Proteins c-mdm2
  • Tumor Suppressor Protein p53 / chemistry
  • Tumor Suppressor Protein p53 / genetics*
  • Tumor Suppressor Protein p53 / metabolism
  • beta-Galactosidase / genetics
  • beta-Galactosidase / metabolism

Substances

  • DNA, Recombinant
  • Leupeptins
  • Multienzyme Complexes
  • Neoplasm Proteins
  • Nuclear Proteins
  • Proto-Oncogene Proteins
  • Tumor Suppressor Protein p53
  • Doxorubicin
  • Chloramphenicol O-Acetyltransferase
  • Mdm2 protein, mouse
  • Proto-Oncogene Proteins c-mdm2
  • beta-Galactosidase
  • Cysteine Endopeptidases
  • Proteasome Endopeptidase Complex
  • benzyloxycarbonylleucyl-leucyl-leucine aldehyde