The MDM2 Antagonist nutlin-3 Sensitizes p53-null Neuroblastoma Cells to Doxorubicin via E2F1 and TAp73

Int J Oncol. 2009 May;34(5):1395-402.

Abstract

Neuroblastoma (NB) is a primitive neuroectodermal tumor and the second most common solid tumor in children. NB exhibits heterogeneous behavior and spontaneous regression can occur in patients under 12 months of age. Response to treatment is both age- and stage-specific; however, patients over 1 year of age are generally considered high risk. NB tumors from these patients are often characterized by alterations in p53 expression and murine double minute (MDM2) activity with concomitant resistance to chemotherapy. We evaluated the ability of nutlin-3 to sensitize a p53-null and doxorubicin-resistant NB cell line, LA155N, to doxorubicin. Nutlin-3 treatment upregulated TAp73 and E2F1 protein levels. It potentiated the ability of doxorubicin to block cell proliferation and activate apoptosis and TAp73 knockdown resulted in a reduction of this sensitization. Additionally, PUMA expression was induced by the combination treatment, but reduced by knockdown of either TAp73 or E2F1. We conclude that, following nutlin-3 treatment, TAp73 and E2F1 are released from MDM2 and activated by doxorubicin to induce PUMA and apoptosis. This study addresses p53-independent mechanisms of nutlin-3 action in chemoresistant NB, especially in combination with chemotherapeutics. We believe that this model has strong clinical relevance for chemoresistant and p53 dysfunctional NB.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antineoplastic Combined Chemotherapy Protocols / pharmacology
  • Antineoplastic Combined Chemotherapy Protocols / therapeutic use
  • Apoptosis / drug effects
  • Apoptosis / genetics
  • Cell Proliferation / drug effects
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / metabolism
  • DNA-Binding Proteins / physiology*
  • Doxorubicin / administration & dosage
  • Doxorubicin / pharmacology*
  • Drug Evaluation, Preclinical
  • Drug Resistance, Neoplasm / drug effects
  • Drug Resistance, Neoplasm / genetics
  • Drug Synergism
  • E2F1 Transcription Factor / metabolism
  • E2F1 Transcription Factor / physiology*
  • Gene Expression Regulation, Neoplastic / drug effects
  • Genes, p53
  • Humans
  • Imidazoles / administration & dosage
  • Imidazoles / pharmacology*
  • Models, Biological
  • Neuroblastoma / drug therapy
  • Neuroblastoma / genetics
  • Neuroblastoma / pathology*
  • Nuclear Proteins / genetics
  • Nuclear Proteins / metabolism
  • Nuclear Proteins / physiology*
  • Piperazines / administration & dosage
  • Piperazines / pharmacology*
  • Proto-Oncogene Proteins c-mdm2 / antagonists & inhibitors*
  • Tumor Cells, Cultured
  • Tumor Protein p73
  • Tumor Suppressor Proteins / genetics
  • Tumor Suppressor Proteins / metabolism
  • Tumor Suppressor Proteins / physiology*

Substances

  • DNA-Binding Proteins
  • E2F1 Transcription Factor
  • E2F1 protein, human
  • Imidazoles
  • Nuclear Proteins
  • Piperazines
  • Tumor Protein p73
  • Tumor Suppressor Proteins
  • nutlin 3
  • Doxorubicin
  • MDM2 protein, human
  • Proto-Oncogene Proteins c-mdm2