The catalytic topoisomerase II inhibitor dexrazoxane induces DNA breaks, ATF3 and the DNA damage response in cancer cells

Br J Pharmacol. 2015 May;172(9):2246-57. doi: 10.1111/bph.13046. Epub 2015 Feb 27.

Abstract

Background and purpose: The catalytic topoisomerase II inhibitor dexrazoxane has been associated not only with improved cancer patient survival but also with secondary malignancies and reduced tumour response.

Experimental approach: We investigated the DNA damage response and the role of the activating transcription factor 3 (ATF3) accumulation in tumour cells exposed to dexrazoxane.

Key results: Dexrazoxane exposure induced topoisomerase IIα (TOP2A)-dependent cell death, γ-H2AX accumulation and increased tail moment in neutral comet assays. Dexrazoxane induced DNA damage responses, shown by enhanced levels of γ-H2AX/53BP1 foci, ATM (ataxia telangiectasia mutated), ATR (ATM and Rad3-related), Chk1 and Chk2 phosphorylation, and by p53 accumulation. Dexrazoxane-induced γ-H2AX accumulation was dependent on ATM. ATF3 protein was induced by dexrazoxane in a concentration- and time-dependent manner, which was abolished in TOP2A-depleted cells and in cells pre-incubated with ATM inhibitor. Knockdown of ATF3 gene expression by siRNA triggered apoptosis in control cells and diminished the p53 protein level in both control and dexrazoxane -treated cells. This was accompanied by increased γ-H2AX accumulation. ATF3 knockdown also delayed the repair of dexrazoxane -induced DNA double-strand breaks.

Conclusions and implications: As with other TOP2A poisons, dexrazoxane induced DNA double-strand breaks followed by activation of the DNA damage response. The DNA damage-triggered ATF3 controlled p53 accumulation and generation of double-strand breaks and is proposed to serve as a switch between DNA damage and cell death following dexrazoxane treatment. These findings suggest a mechanistic explanation for the diverse clinical observations associated with dexrazoxane.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Activating Transcription Factor 3 / genetics
  • Activating Transcription Factor 3 / metabolism*
  • Antigens, Neoplasm / genetics
  • Antigens, Neoplasm / metabolism
  • Apoptosis / drug effects
  • Ataxia Telangiectasia Mutated Proteins / antagonists & inhibitors
  • Ataxia Telangiectasia Mutated Proteins / metabolism
  • Cell Line, Tumor
  • DNA Breaks, Double-Stranded*
  • DNA Topoisomerases, Type II / genetics
  • DNA Topoisomerases, Type II / metabolism
  • DNA-Binding Proteins / antagonists & inhibitors*
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / metabolism
  • Dexrazoxane / pharmacology*
  • Dose-Response Relationship, Drug
  • Fibrosarcoma / drug therapy*
  • Fibrosarcoma / genetics
  • Fibrosarcoma / metabolism
  • Fibrosarcoma / pathology
  • Histones / metabolism
  • Humans
  • Intracellular Signaling Peptides and Proteins / metabolism
  • Poly-ADP-Ribose Binding Proteins
  • RNA Interference
  • Signal Transduction / drug effects
  • Time Factors
  • Topoisomerase II Inhibitors / pharmacology*
  • Transfection
  • Tumor Suppressor Protein p53 / metabolism
  • Tumor Suppressor p53-Binding Protein 1

Substances

  • ATF3 protein, human
  • Activating Transcription Factor 3
  • Antigens, Neoplasm
  • DNA-Binding Proteins
  • H2AX protein, human
  • Histones
  • Intracellular Signaling Peptides and Proteins
  • Poly-ADP-Ribose Binding Proteins
  • TP53 protein, human
  • TP53BP1 protein, human
  • Topoisomerase II Inhibitors
  • Tumor Suppressor Protein p53
  • Tumor Suppressor p53-Binding Protein 1
  • Dexrazoxane
  • ATM protein, human
  • Ataxia Telangiectasia Mutated Proteins
  • DNA Topoisomerases, Type II
  • TOP2A protein, human