Estrogen receptor α (ERα) status evaluation using RNAscope in situ hybridization: a reliable and complementary method for IHC in breast cancer tissues

Hum Pathol. 2017 Mar:61:121-129. doi: 10.1016/j.humpath.2016.12.005. Epub 2016 Dec 16.

Abstract

Estrogen receptor α (ERα) plays a significant role in the development of breast cancer and has been used clinically as an endocrine therapeutic target. Currently, clinical laboratories use immunohistochemistry (IHC) to determine the ERα status of patients in order to distinguish those who would benefit from endocrine therapy. This method is highly subjective, requires a large amount of tumor tissue, and may generate false-negative results. To improve the detection of ERα, we used a new RNA in situ hybridization technique (RNAscope) and compared its use with IHC in 72 breast cancer tissues (47 ERα positive and 25 ERα negative). Then we evaluated ERα mRNA by RT-qPCR with RNAscope. An unobvious difference was found between reverse-transcription quantitative polymerase chain reaction (RT-qPCR) and IHC, but a positive correlation was found between RNAscope and IHC. In addition, breast cancer is a highly heterogeneous cancer, and RNAscope could easily reveal the heterogeneity in breast cancer. Moreover, we found that some ERα IHC-based negative and RNAscope-based positive test results were detected as positive after testing with IHC again. Our findings suggest that RNAscope may be a complementary method for improving the detection of patient ERα status and has potential clinical utility.

Keywords: Breast cancer; Estrogen receptor α; Heterogeneity; Immunohistochemistry; RNAscope; Real-time RT-PCR.

Publication types

  • Comparative Study

MeSH terms

  • Adult
  • Biomarkers, Tumor / analysis*
  • Biomarkers, Tumor / genetics*
  • Biopsy
  • Breast Neoplasms / chemistry*
  • Breast Neoplasms / genetics*
  • Breast Neoplasms / pathology
  • Estrogen Receptor alpha / analysis*
  • Estrogen Receptor alpha / genetics*
  • Female
  • Humans
  • Immunohistochemistry*
  • In Situ Hybridization / methods*
  • Middle Aged
  • Predictive Value of Tests
  • RNA, Messenger / genetics*
  • Real-Time Polymerase Chain Reaction
  • Reproducibility of Results
  • Reverse Transcriptase Polymerase Chain Reaction
  • Tissue Array Analysis

Substances

  • Biomarkers, Tumor
  • ESR1 protein, human
  • Estrogen Receptor alpha
  • RNA, Messenger