Quantitative assessment of HER2 gene amplification of breast cancer using droplet digital PCR

Pathol Int. 2021 Aug;71(8):538-547. doi: 10.1111/pin.13128. Epub 2021 Jul 21.

Abstract

We previously reported the usefulness of droplet digital polymerase chain reaction (ddPCR) for the assessment of Human epithelial growth factor receptor 2 (HER2) gene amplification in breast cancer using formalin-fixed and paraffin-embedded sections. In our previous study, we combined HER2/CEP17 ratio (HER2 gene signals to chromosome 17 signals) with ddPCR and tumor content ratio (TCR) of each sample and determined the HER2 status by adopting a two-dimensional chart. This "ddPCR-TCR method" showed a high concordance with conventional HER2 status. In this study, we updated our method to assess the HER2 status of breast cancer in a more quantitative manner. We combined obtained data of the ddPCR ratio [Rx ] and TCR [x]; we calculated "(Rx - 1)/x + 1" for 41 samples with primary breast cancer and named the value led by this formula as "eHER2 (estimated HER2/CEP17 ratio of a tumor cell)". eHER2 was equivalent to conventional in situ hybridization (ISH) HER2/CEP17 ratio in most cases. eHER2 and ISH ratio showed a strong correlation (Spearman rank correlation ρ = 0.70, p < 0.0001). The obtained results indicated that eHER2 is a potential tool for HER2 status diagnosis in breast cancer.

Keywords: HER2 genes; breast neoplasms; diagnostic technics and procedures; gene amplification; polymerase chain reaction.

MeSH terms

  • Adult
  • Aged
  • Aged, 80 and over
  • Breast Neoplasms* / diagnosis
  • Breast Neoplasms* / genetics
  • Breast Neoplasms* / metabolism
  • Breast Neoplasms* / pathology
  • Female
  • Genes, erbB-2
  • Humans
  • In Situ Hybridization, Fluorescence
  • Middle Aged
  • Oncogene Proteins v-erbB / genetics*
  • Paraffin Embedding
  • Pathology, Molecular
  • Polymerase Chain Reaction / methods*

Substances

  • Oncogene Proteins v-erbB