Differential epithelium DNA damage response to ATM and DNA-PK pathway inhibition in human prostate tissue culture

Cell Cycle. 2011 Oct 15;10(20):3545-53. doi: 10.4161/cc.10.20.17841. Epub 2011 Oct 15.


The ability of cells to respond and repair DNA damage is fundamental for the maintenance of genomic integrity. Ex vivo culturing of surgery-derived human tissues has provided a significant advancement to assess DNA damage response (DDR) in the context of normal cytoarchitecture in a non-proliferating tissue. Here, we assess the dependency of prostate epithelium DDR on ATM and DNA-PKcs, the major kinases responsible for damage detection and repair by nonhomologous end-joining (NHEJ), respectively. DNA damage was caused by ionizing radiation (IR) and cytotoxic drugs, cultured tissues were treated with ATM and DNA-PK inhibitors, and DDR was assessed by phosphorylation of ATM and its targets H2AX and KAP1, a heterochromatin binding protein. Phosphorylation of H2AX and KAP1 was fast, transient and fully dependent on ATM, but these responses were moderate in luminal cells. In contrast, DNA-PKcs was phosphorylated in both luminal and basal cells, suggesting that DNA-PK-dependent repair was also activated in the luminal cells despite the diminished H2AX and KAP1 responses. These results indicate that prostate epithelial cell types have constitutively dissimilar responses to DNA damage. We correlate the altered damage response to the differential chromatin state of the cells. These findings are relevant in understanding how the epithelium senses and responds to DNA damage.

Publication types

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

MeSH terms

  • Ataxia Telangiectasia Mutated Proteins
  • Cell Cycle Proteins / metabolism*
  • Cells, Cultured
  • DNA Damage*
  • DNA End-Joining Repair / physiology*
  • DNA-Activated Protein Kinase / metabolism*
  • DNA-Binding Proteins / metabolism*
  • Epithelium / metabolism*
  • Histones / metabolism
  • Humans
  • Image Processing, Computer-Assisted
  • Immunohistochemistry
  • Male
  • Microscopy, Confocal
  • Microscopy, Fluorescence
  • Phosphorylation
  • Prostate / cytology
  • Protein-Serine-Threonine Kinases / metabolism*
  • Radiation, Ionizing
  • Repressor Proteins / metabolism
  • Signal Transduction / physiology*
  • Tripartite Motif-Containing Protein 28
  • Tumor Suppressor Proteins / metabolism*


  • Cell Cycle Proteins
  • DNA-Binding Proteins
  • H2AX protein, human
  • Histones
  • Repressor Proteins
  • Tumor Suppressor Proteins
  • TRIM28 protein, human
  • Tripartite Motif-Containing Protein 28
  • ATM protein, human
  • Ataxia Telangiectasia Mutated Proteins
  • DNA-Activated Protein Kinase
  • Protein-Serine-Threonine Kinases