Real-time PCR: detection of oestrogen receptor-alpha and -beta isoforms and variants in breast cancer

Anticancer Res. 2010 Oct;30(10):4147-56.

Abstract

Background: Splice variants exist for both alpha and beta oestrogen receptors (ERs). Oestrogen function results from a balance between the wild-type ERs (wt) and their variants.

Patients and methods: Forty formalin-fixed paraffin-embedded breast cancer samples were analysed by real-time PCR using ERα primer sets detecting wt and exon-deleted 3, 5, 6 and 7 variants. The ERβ primer sets detected wt ERβ1 and ERβ2 and ERβ5 variants. At the end of the PCR cycles, a dissociation curve was generated showing the peaks for each sample at specific melting temperatures (Tm); finding more than one peak indicated the presence of variants.

Results: Many samples expressed both wt ER isoforms and their variants. The Tm value served as a cut-off point for determination of wt versus variant ER expression.

Conclusion: This method of detection of wt and variant ER could help in patient selection for anti-oestrogen therapy and in monitoring response to therapy.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Breast Neoplasms / genetics
  • Breast Neoplasms / metabolism*
  • Cell Line, Tumor
  • Estrogen Receptor alpha / biosynthesis*
  • Estrogen Receptor alpha / genetics
  • Estrogen Receptor beta / biosynthesis*
  • Estrogen Receptor beta / genetics
  • Female
  • Gene Amplification
  • Humans
  • Paraffin Embedding
  • Reverse Transcriptase Polymerase Chain Reaction / methods*

Substances

  • Estrogen Receptor alpha
  • Estrogen Receptor beta