Accurate detection and quantification of epigenetic and genetic second hits in BRCA1 and BRCA2-associated hereditary breast and ovarian cancer reveals multiple co-acting second hits

Cancer Lett. 2018 Jul 1:425:125-133. doi: 10.1016/j.canlet.2018.03.026. Epub 2018 Mar 23.

Abstract

Background: This study characterizes the second hit spectrum in BRCA1 and BRCA2-associated breast and ovarian cancers at both gene loci to investigate if second hit mechanisms are mutually exclusive or able to coincide within the same tumor.

Methods: Loss of heterozygosity, somatic point mutations and copy number alterations along with promoter methylation were studied in 56 breast and 15 ovarian cancers from BRCA1 and BRCA2 germline mutation carriers. A mathematical methodology was introduced to quantify the tumor cell population carrying a second hit.

Results: Copy neutral LOH was the most prevalent LOH mechanism in this cohort (BC 69%, OC 67%). However, only 36% of BC and 47% of OC showed LOH in all cancerous cells. Somatic intragenic deletions and methylated subclones were also found in combination with (partial) loss of heterozygosity. Unequivocal deleterious somatic point mutations were not identified in this cohort.

Conclusion: Different mechanisms inactivating the wild type allele are present within the same tumor sample at various extents. Results indicate that BRCA1/2-linked breast and ovarian cancer cells are predominantly characterized by LOH, but harbor a complex combination of second hits at various frequencies.

Keywords: BRCA1; BRCA2; Loss of heterozygosity; Methylation; Tumor cell percentage.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • BRCA1 Protein / genetics*
  • BRCA2 Protein / genetics*
  • Cohort Studies
  • DNA Copy Number Variations
  • DNA Methylation
  • Epigenesis, Genetic
  • Female
  • Germ-Line Mutation
  • Hereditary Breast and Ovarian Cancer Syndrome / genetics*
  • Humans
  • Loss of Heterozygosity
  • Models, Theoretical
  • Point Mutation
  • Promoter Regions, Genetic
  • Sequence Analysis, DNA / methods*

Substances

  • BRCA1 Protein
  • BRCA1 protein, human
  • BRCA2 Protein
  • BRCA2 protein, human