Kinetochore analysis of micronuclei allows insights into the actions of colcemid and mitomycin C

Mutat Res. 1991 Sep;261(1):57-68. doi: 10.1016/0165-1218(91)90098-7.

Abstract

We have induced micronuclei in two strains of diploid human fibroblasts with a known aneugen, colcemid, and a known clastogen, mitomycin C. Using immunofluorescence to detect the presence of kinetochores in micronuclei, we were able to demonstrate a 26.8-fold increase in fluorescence-positive micronuclei (aneuploidy) in colcemid-treated cells. However, colcemid also induced an increase in kinetochore-negative micronuclei. Our findings support previous reports that suggest colcemid may induce chromosome breakage in addition to its major aneugenic effect. The frequency of kinetochore-negative micronuclei (chromosome breakage) in mitomycin C-treated cells rose an average of 7.9-fold in the two test strains, a clear reflection of its clastogenic action. However, a 4-fold increase in the kinetochore-positive fraction was seen. We conclude that the fibroblast micronucleus assay, coupled with kinetochore immunofluorescence, provides a useful screening approach for genotoxic agents. The delineation of the precise mechanism by which an agent perturbs the rates of chromosomal breakage or lag may require more detailed analysis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aneuploidy
  • Chromosome Aberrations
  • Demecolcine / pharmacology*
  • Evaluation Studies as Topic
  • Fibroblasts / ultrastructure
  • Fluorescent Antibody Technique
  • Humans
  • Micronuclei, Chromosome-Defective / drug effects*
  • Mitomycin
  • Mitomycins / pharmacology*
  • Mutagenicity Tests*

Substances

  • Mitomycins
  • Mitomycin
  • Demecolcine