Effect of a poly(ADP-ribose) polymerase-1 inhibitor against esophageal squamous cell carcinoma cell lines

Cancer Sci. 2014 Feb;105(2):202-10. doi: 10.1111/cas.12322. Epub 2013 Dec 5.

Abstract

Effective molecular target drugs that improve therapeutic efficacy with fewer adverse effects for esophageal cancer are highly anticipated. Poly(ADP-ribose) polymerase (PARP) inhibitors have been proposed as low-toxicity agents to treat double strand break (DSB)-repair defective tumors. Several findings imply the potential relevance of DSB repair defects in the tumorigenesis of esophageal squamous cell carcinoma (ESCC). We evaluated the effect of a PARP Inhibitor (AZD2281) on the TE-series ESCC cell lines. Of these eight cell lines, the clonogenic survival of one (TE-6) was reduced by AZD2281 to the level of DSB repair-defective Capan-1 and HCC1937 cells. AZD2281-induced DNA damage was implied by increases in γ-H2AX and cell cycle arrest at G2/M phase. The impairment of DSB repair in TE-6 cells was suggested by a sustained increase in γ-H2AX levels and the tail moment calculated from a neutral comet assay after X-ray irradiation. Because the formation of nuclear DSB repair protein foci was impaired in TE-6 cells, whole-exome sequencing of these cells was performed to explore the gene mutations that might be responsible. A novel mutation in RNF8, an E3 ligase targeting γ-H2AX was identified. Consistent with this, polyubiquitination of γ-H2AX after irradiation was impaired in TE-6 cells. Thus, AZD2281 induced growth retardation of the DSB repair-impaired TE-6 cells. Interestingly, a strong correlation between basal expression levels of γ-H2AX and sensitivity to AZD2281was observed in the TE-series cells (R(2) = 0.5345). Because the assessment of basal DSB status could serve as a biomarker for selecting PARP inhibitor-tractable tumors, further investigation is warranted.

Keywords: DNA repair; RNF8; esophageal cancer; poly(ADP-ribose) polymerase inhibitor; γ-H2AX.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Carcinoma, Squamous Cell / drug therapy*
  • Carcinoma, Squamous Cell / enzymology
  • Carcinoma, Squamous Cell / genetics
  • Carcinoma, Squamous Cell / metabolism
  • Cell Cycle Checkpoints / drug effects
  • Cell Division / drug effects
  • Cell Line, Tumor
  • DNA Damage / drug effects
  • DNA Repair / drug effects
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / metabolism
  • Esophageal Neoplasms / drug therapy*
  • Esophageal Neoplasms / enzymology
  • Esophageal Neoplasms / genetics
  • Esophageal Neoplasms / metabolism
  • Esophageal Squamous Cell Carcinoma
  • G2 Phase / drug effects
  • Histones / genetics
  • Histones / metabolism
  • Humans
  • MCF-7 Cells
  • Molecular Sequence Data
  • Mutation / drug effects
  • Mutation / genetics
  • Phthalazines / adverse effects
  • Phthalazines / pharmacology*
  • Piperazines / adverse effects
  • Piperazines / pharmacology*
  • Poly(ADP-ribose) Polymerase Inhibitors*
  • Ubiquitin-Protein Ligases

Substances

  • DNA-Binding Proteins
  • H2AX protein, human
  • Histones
  • Phthalazines
  • Piperazines
  • Poly(ADP-ribose) Polymerase Inhibitors
  • RNF8 protein, human
  • Ubiquitin-Protein Ligases
  • olaparib

Associated data

  • GENBANK/AG014699