Cell proliferation as a requirement for development of the contact effect in Chinese hamster V79 spheroids

Radiat Res. 1989 Jan;117(1):79-92.


Chinese hamster V79 cells grown for several hours in suspension culture form spheroids which are more resistant to killing by ionizing radiation than cells grown on petri dishes, a phenomenon known as the "contact effect." Previous results using the alkali-unwinding assay as a measure of DNA damage have implicated differences in DNA conformation as contributing to this effect; spheroid DNA denatures more slowly in dilute alkali than monolayer DNA, perhaps due to the presence of constraints to DNA unwinding. In this paper, the rate of development of radiation resistance is shown to be similar when either cell survival or DNA unwinding is used as an end point. At the midpoint for development of resistance, approximately 10 h, the unwinding kinetics indicate that either half of the cells contain constraints to DNA unwinding, or half of the DNA in all of the cells contains constraints. The latter explanation appears more likely since all cells seem to develop these constraints at the same rate, regardless of position in the cell cycle or the degree of contact with other cells. Results using the microelectrophoresis assay to measure damage to individual nuclei confirm the fact that 10-h cultures show a homogeneous radiation response intermediate between that of monolayers and spheroids. Incubation of cells at room temperature or in the presence of drugs which inhibit cell cycle progression prevents full development of the contact effect. Conversely, incubation of cells in medium containing inhibitors of polyamine synthesis, adenylcyclase, glutathione synthesis, poly(ADP-ribose)polymerase, topoisomerase II, or cell-cell communication does not inhibit development of the contact effect as measured by DNA-unwinding kinetics.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Aggregation*
  • Cell Communication
  • Cell Cycle
  • Cell Division
  • Cell Line
  • Cell Survival / radiation effects
  • DNA / radiation effects
  • DNA Damage
  • Demecolcine / pharmacology
  • Nucleic Acid Conformation / radiation effects
  • Radiation Tolerance*


  • DNA
  • Demecolcine