Analysis of epidermal growth factor receptor mRNA expression by polymerase chain reaction assay in 94 human breast adenocarcinoma tumors

Breast Cancer Res Treat. 1994;30(3):275-82. doi: 10.1007/BF00665968.

Abstract

It is well known that breast cancer cells can synthesize and secrete various growth factors that are able to stimulate tumor growth through autocrine and/or paracrine mechanisms. EGF is one of these growth factors involved in normal breast epithelial development and tumor proliferation. EGF and TGF alpha (EGF-like peptide) are produced in variable amounts and both bind to the EGF receptor (EGF-R). Previous investigation in the laboratory measuring free and occupied EGF-R sites by differential ligand binding assays had demonstrated that non-occupied and total binding sites were present in 54 and 90% of 216 breast tumor biopsies respectively. EGF-R appeared to be totally masked by endogenous ligand in 40 and 21% of estrogen receptor positive and negative tumors respectively. The aim of the present study was to check by a molecular method the expression of the EGF-R gene. The PCR method was applied to 94 tumor samples of the previous series. Total RNA was treated with 0.5 units of Rnase-free Dnase/mg of RNA to remove any contaminating DNA. We simultaneously reverse transcribed and amplified another transcript (beta-actin) as an internal standard. Both signals were present in 88 of the 94 samples while the presence of EGF-R was detected in 74 of them when assessed by radioligand assay. The findings indicate that 93% of the tumors analysed in this series expressed EGF-R mRNA, in agreement with our previous data on occupied EGF-R sites, i.e. two-fold more than by using the standard binding assay. No significant correlation was observed between the expression of the EGF-R gene and the estrogen receptor content.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenocarcinoma / metabolism*
  • Adenocarcinoma / pathology
  • Base Sequence
  • Biopsy
  • Blotting, Northern
  • Breast Neoplasms / metabolism*
  • Breast Neoplasms / pathology
  • DNA Primers
  • Electrophoresis, Agar Gel
  • ErbB Receptors / analysis
  • ErbB Receptors / biosynthesis*
  • Gene Expression*
  • Humans
  • Molecular Sequence Data
  • Polymerase Chain Reaction / methods*
  • RNA, Messenger / analysis
  • RNA, Messenger / biosynthesis*
  • Radioligand Assay

Substances

  • DNA Primers
  • RNA, Messenger
  • ErbB Receptors