Analyses of brain tumor cell lines confirm a simple model of relationships among fluorescence in situ hybridization, DNA index, and comparative genomic hybridization

Genes Chromosomes Cancer. 1997 Dec;20(4):311-9. doi: 10.1002/(sici)1098-2264(199712)20:4<311::aid-gcc1>3.0.co;2-4.

Abstract

Several techniques are commonly used for genetic analysis of interphase nuclei. Flow cytometry assays the distribution of DNA content in populations of nuclei stained with a DNA-specific fluorochrome. Fluorescence in situ hybridization (FISH) quantifies the number of copies of a specific DNA sequence in single nuclei. Comparative genomic hybridization (CGH) assesses the relative copy number of DNA sequences throughout a test genome by comparing the signal intensities of test and reference DNA samples hybridized to a template of normal metaphase chromosomes. In principle, there are specific relationship among data obtained from these measurements, and combined measurements should provide a more comprehensive view of the sample that is analyzed. We applied these three techniques to nine brain tumor cell lines and find that a model of CGH that includes unsuppressed repeat sequences describes the data well. We estimate that up to 35% of the fluorescence intensity in well-blocked CGH preparations may not represent unique sequences. Taking these factors into account, our results are, in general, mutually consistent, and highlight issues critical for interpreting CGH preparations.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Algorithms
  • Animals
  • Brain Neoplasms / genetics*
  • DNA Probes
  • DNA, Neoplasm / analysis*
  • Flow Cytometry
  • Gene Dosage
  • Humans
  • In Situ Hybridization, Fluorescence / methods*
  • Models, Genetic*
  • Nucleic Acid Hybridization / methods*
  • Tumor Cells, Cultured

Substances

  • DNA Probes
  • DNA, Neoplasm