Drug-induced death of leukaemic cells after G2/M arrest: higher order DNA fragmentation as an indicator of mechanism

Br J Cancer. 1998;77(1):40-50. doi: 10.1038/bjc.1998.7.

Abstract

Many reports have documented apoptotic death in different cell types within hours of exposure to cytotoxic drugs; lower drug concentrations may cause cell cycle arrest at G2/M and subsequent death, which has been distinguished from 'classic' apoptosis. We have analysed etoposide-induced cell death in two lymphoblastoid T-cell lines, CCRF-CEM and MOLT-4, specifically in relation to DNA cleavage as indicated by pulse-field gel and conventional electrophoresis. High (5 microM) concentration etoposide causes 50-kb cleavage of DNA that occurs at the same time as apoptotic morphology and internucleosomal cleavage. At lower concentrations (0.5-0.05 microM), sequential change may be discerned with altered gene expression being similar to that at high dose, but preceding cell cycle arrest and 50-kb cleavage. These last changes, in turn, clearly precede internucleosomal fragmentation of DNA, vital dye staining and morphological evidence cell death. The pattern of higher order fragmentation constitutes a sensitive indicator of commitment to cell death in these cells. Morphological evidence of cell death is associated with internucleosomal fragmentation in one of the lines, but the pattern of 50-kb DNA cleavage provides the clearest evidence of commonality in death processes occurring at low and high drug concentration.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antineoplastic Agents, Phytogenic / pharmacology*
  • Apoptosis / drug effects
  • Cell Division / drug effects
  • Cell Size / drug effects
  • Coloring Agents
  • DNA Fragmentation*
  • DNA, Neoplasm*
  • Electrophoresis, Gel, Pulsed-Field
  • Etoposide / pharmacology*
  • G2 Phase / drug effects*
  • Gene Expression / drug effects
  • Humans
  • Leukemia / genetics
  • Leukemia / pathology*
  • Mitosis / drug effects*
  • Polymerase Chain Reaction
  • Trypan Blue
  • Tumor Cells, Cultured / drug effects

Substances

  • Antineoplastic Agents, Phytogenic
  • Coloring Agents
  • DNA, Neoplasm
  • Etoposide
  • Trypan Blue