Laser microdissection and microsatellite analyses of breast cancer reveal a high degree of tumor heterogeneity

Pathobiology. 2000;68(4-5):180-90. doi: 10.1159/000055921.

Abstract

Carcinomas with productive fibrosis are the most common forms of breast cancer. Analysis of tumor-specific genomic alterations can be compromised by the presence of normal cells, demanding microdissection of small tumor areas to detect loss of heterozygosity (LOH) and microsatellite instability (MSI). The aim of this study was to evaluate the importance of precise laser microdissection for microsatellite analyses and investigation of tumor heterogeneity in breast cancer. 39 primary breast tumor samples were analyzed for MSI and LOH by PCR followed by polyacrylamide gel electrophoresis and silver staining using 15 microsatellite markers. Different tumor areas were processed separately in 30 patients. Both intraductal and invasive breast cancer regions were investigated in 11 patients. The following results were obtained: (1) accurate microdissection revealed MSI in 3 or more of the investigated markers (> or =20%) in 33% of the patients, a higher frequency than reported previously; (2) laser microdissection was 43% more sensitive in detection of LOH compared to manual microdissection due to a reduction of contamination by normal cells, and (3) 29 of 30 investigated tumors showed heterogeneity of genetic alterations in different tumor regions. Laser-based microdissection is a valuable tool in genetic analysis of desmoplastic tumors and allows an accurate determination of genetic alterations in histologically different tumor regions.

Publication types

  • Comparative Study

MeSH terms

  • Adult
  • Breast Neoplasms / genetics*
  • Breast Neoplasms / pathology
  • Carcinoma, Ductal, Breast / genetics
  • Carcinoma, Ductal, Breast / pathology
  • Cell Separation / instrumentation
  • Cell Separation / methods
  • Chromosomes, Human, Pair 17 / genetics
  • Dissection / instrumentation
  • Dissection / methods*
  • Female
  • Genetic Heterogeneity*
  • Humans
  • Lasers*
  • Loss of Heterozygosity
  • Microsatellite Repeats / genetics*
  • Middle Aged
  • Predictive Value of Tests
  • Prospective Studies
  • Trinucleotide Repeat Expansion / genetics