Deoxynucleoside salvage facilitates DNA repair during ribonucleotide reductase blockade in human cervical cancers

Radiat Res. 2011 Oct;176(4):425-33. doi: 10.1667/rr2556.1. Epub 2011 Jul 14.


Cells generate 2'-deoxyribonucleoside triphosphates (dNTPs) for both replication and repair of damaged DNA predominantly through de novo reduction of intracellular ribonucleotides by ribonucleotide reductase (RNR). Cells can also salvage deoxynucleosides by deoxycytidine kinase/thymidine kinase 1 in the cytosol or by deoxyguanosine kinase/thymidine kinase 2 in mitochondria. In this study we investigated whether the salvage dNTP supply pathway facilitates DNA damage repair, promoting cell survival, when pharmacological inhibition of RNR by 3-aminopyridine-2-carboxaldehyde thiosemicarbazone (3-AP, NSC no. 663249) impairs the de novo pathway. Human cervical cancer cells were subjected to radiation with or without 3-AP under medium deoxynucleoside concentrations of 0, 0.05, 0.5 and 5.0 µM. Efficacy of DNA damage repair was assessed by γ-H2AX flow cytometry and focus counts, by single cell electrophoresis (Comet assay), and by caspase 3 cleavage assay as a marker of treatment-induced apoptosis. Cell survival was assessed by colony formation. We found that deoxyribonucleotide salvage facilitates DNA repair during RNR inhibition by 3-AP and that salvage reduces the radiochemosensitivity of human cervical cancer cells.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Culture Media / pharmacology
  • DNA Repair / drug effects*
  • DNA Repair / radiation effects*
  • Deoxyribonucleosides / metabolism*
  • Deoxyribonucleosides / pharmacology*
  • Deoxyribonucleotides / metabolism
  • Enzyme Inhibitors / pharmacology*
  • Female
  • Humans
  • Oxidation-Reduction / drug effects
  • Oxidation-Reduction / radiation effects
  • Pyridines / pharmacology
  • Radiation Tolerance / drug effects
  • Radiation Tolerance / radiation effects
  • Ribonucleotide Reductases / antagonists & inhibitors*
  • Thiosemicarbazones / pharmacology
  • Uterine Cervical Neoplasms / enzymology
  • Uterine Cervical Neoplasms / genetics*
  • Uterine Cervical Neoplasms / metabolism


  • Culture Media
  • Deoxyribonucleosides
  • Deoxyribonucleotides
  • Enzyme Inhibitors
  • Pyridines
  • Thiosemicarbazones
  • 3-aminopyridine-2-carboxaldehyde thiosemicarbazone
  • Ribonucleotide Reductases