Quantitative analysis of her-2/neu (Erbb2) gene expression using reverse transcriptase polymerase chain reaction

Fazlul H. Sarkar, Daniel W. Visscher, John D. Crissman

Research output: Contribution to journalArticlepeer-review

12 Scopus citations

Abstract

Inappropriate expression of Her-2/neu (ERBB2) gene has been associated with impaired breast cancer prognosis, suggesting a functional role in tumor progression. Herein we describe a quantitative method for analysis of Her-2/neu gene messenger RNA (mRNA), which employs reverse transcriptase polymerase chain reaction (RT-PCR) on a 10-xm cryostat section. The technique combines modified RNA extraction with complementary DNA (cDNA) synthesis to achieve a high level of sensitivity. Utilizing this PCR-based gene expression assay, we were able to quantitate variable amounts of Her-2/neu mRNA in cell lines with established levels of gene expression and in clinical human breast cancer specimens. In clinical samples, mRNA levels correlated with intensity of immu-noperoxidase staining for corresponding oncoprotein. We conclude that PCR-based mRNA quantitation can be applied to quantitative analysis of Her-2/neu gene expression, and potentially many other genes, in samples of limited size.

Original languageEnglish (US)
Pages (from-to)210-218
Number of pages9
JournalDiagnostic Molecular Pathology
Volume2
Issue number1
DOIs
StatePublished - Jan 1 1993

Keywords

  • Breast carcinoma
  • Erbb2
  • Polymerase chain reaction

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Molecular Biology
  • Cell Biology

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