Adenoviral-E2F-1 radiosensitizes p53wild-type and p53null human prostate cancer cells

Int J Radiat Oncol Biol Phys. 2005 Sep 1;63(1):238-46. doi: 10.1016/j.ijrobp.2005.04.033.

Abstract

Purpose: E2F-1 is a transcription factor that enhances the radiosensitivity of various cell lines by inducing apoptosis. However, there are conflicting data concerning whether this enhancement is mediated via p53 dependent pathways. Additionally, the role of E2F-1 in the response of human prostate cancer to radiation has not been well characterized. In this study, we investigated the effect of Adenoviral-E2F-1 (Ad-E2F-1) on the radiosensitivity of p53wild-type (LNCaP) and p53null (PC3) prostate cancer cell lines.

Methods and materials: LNCaP and PC3 cells were transduced with Ad-E2F-1, Adenoviral-Luciferase (Ad-Luc) control vector, or Adenoviral-p53 (Ad-p53). Expression of E2F-1 and p53 was examined by Western blot analysis. Annexin V and caspase 3 + 7 assays were performed to estimate the levels of apoptosis. Clonogenic survival assays were used to determine overall cell death. Statistical significance was determined by analysis of variance, using the Bonferroni method to correct for multiple comparisons.

Results: Western blot analysis confirmed the efficacy of transductions with Ad-E2F-1 and Ad-p53. Ad-E2F-1 transduction significantly enhanced apoptosis and decreased clonogenic survival in both cell lines. These effects were compounded by the addition of RT. Although E2F-1-mediated radiosensitization was independent of p53 status, this effect was more pronounced in p53wild-type LNCaP cells. When PC3 cells were treated with Ad-p53 in combination with RT and Ad-E2F-1, there was at least an additive reduction in clonogenic survival.

Conclusions: Our results suggest that Ad-E2F-1 significantly enhances the response of p53wild-type and p53null prostate cancer cells to radiation therapy, although radiosensitization is more pronounced in the presence of p53. Ad-E2F-1 may be a useful adjunct to radiation therapy in the treatment of prostate cancer.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adenoviridae
  • Apoptosis / genetics
  • Apoptosis / radiation effects
  • Cell Cycle Proteins / metabolism
  • Cell Cycle Proteins / therapeutic use*
  • Cell Line, Tumor / radiation effects
  • DNA-Binding Proteins / metabolism
  • DNA-Binding Proteins / therapeutic use*
  • E2F Transcription Factors
  • E2F1 Transcription Factor
  • Genes, p53*
  • Humans
  • Male
  • Prostatic Neoplasms / genetics*
  • Prostatic Neoplasms / metabolism
  • Prostatic Neoplasms / radiotherapy*
  • Proto-Oncogene Proteins p21(ras) / metabolism
  • Radiation Tolerance
  • Radiation-Sensitizing Agents / therapeutic use*
  • Transcription Factors / metabolism
  • Transcription Factors / therapeutic use*
  • Transduction, Genetic / methods*
  • Tumor Stem Cell Assay / methods
  • Tumor Suppressor Protein p53 / metabolism

Substances

  • Cell Cycle Proteins
  • DNA-Binding Proteins
  • E2F Transcription Factors
  • E2F1 Transcription Factor
  • E2F1 protein, human
  • Radiation-Sensitizing Agents
  • Transcription Factors
  • Tumor Suppressor Protein p53
  • HRAS protein, human
  • Proto-Oncogene Proteins p21(ras)