DNA-PKcs Inhibition Sensitizes Human Chondrosarcoma Cells to Carbon Ion Irradiation via Cell Cycle Arrest and Telomere Capping Disruption

Int J Mol Sci. 2024 Jun 4;25(11):6179. doi: 10.3390/ijms25116179.

Abstract

In order to overcome the resistance to radiotherapy in human chondrosarcoma cells, the prevention from efficient DNA repair with a combined treatment with the DNA-dependent protein kinase catalytic subunit (DNA-PKcs) inhibitor AZD7648 was explored for carbon ion (C-ion) as well as reference photon (X-ray) irradiation (IR) using gene expression analysis, flow cytometry, protein phosphorylation, and telomere length shortening. Proliferation markers and cell cycle distribution changed significantly after combined treatment, revealing a prominent G2/M arrest. The expression of the G2/M checkpoint genes cyclin B, CDK1, and WEE1 was significantly reduced by IR alone and the combined treatment. While IR alone showed no effects, additional AZD7648 treatment resulted in a dose-dependent reduction in AKT phosphorylation and an increase in Chk2 phosphorylation. Twenty-four hours after IR, the key genes of DNA repair mechanisms were reduced by the combined treatment, which led to impaired DNA repair and increased radiosensitivity. A time-dependent shortening of telomere length was observed in both cell lines after combined treatment with AZD7648 and 8 Gy X-ray/C-ion IR. Our data suggest that the inhibition of DNA-PKcs may increase sensitivity to X-rays and C-ion IR by impairing its functional role in DNA repair mechanisms and telomere end protection.

Keywords: AZD7648; DNA-PKcs inhibitor; carbon ion irradiation; chondrosarcoma; telomere length.

MeSH terms

  • Bone Neoplasms / drug therapy
  • Bone Neoplasms / genetics
  • Bone Neoplasms / metabolism
  • Bone Neoplasms / pathology
  • Cell Cycle Checkpoints / drug effects
  • Cell Cycle Checkpoints / radiation effects
  • Cell Line, Tumor
  • Cell Proliferation / drug effects
  • Chondrosarcoma* / drug therapy
  • Chondrosarcoma* / genetics
  • Chondrosarcoma* / metabolism
  • Chondrosarcoma* / radiotherapy
  • DNA Repair / drug effects
  • DNA-Activated Protein Kinase* / antagonists & inhibitors
  • DNA-Activated Protein Kinase* / genetics
  • DNA-Activated Protein Kinase* / metabolism
  • G2 Phase Cell Cycle Checkpoints / drug effects
  • G2 Phase Cell Cycle Checkpoints / radiation effects
  • Heavy Ion Radiotherapy*
  • Humans
  • Pyrazoles / pharmacology
  • Radiation Tolerance / drug effects
  • Telomere* / drug effects
  • Telomere* / metabolism

Substances

  • DNA-Activated Protein Kinase
  • PRKDC protein, human
  • Pyrazoles