Quantitative analysis of Her-2/neu (ERBB2) gene expression using reverse transcriptase polymerase chain reaction

F. H. Sarkar, Daniel W Visscher, J. D. Crissman

Research output: Contribution to journalArticle

12 Citations (Scopus)

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-μm 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 immunoperoxidase 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 number3
StatePublished - 1993
Externally publishedYes

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erbB-2 Genes
Reverse Transcriptase Polymerase Chain Reaction
Gene Expression
Messenger RNA
Breast Neoplasms
Polymerase Chain Reaction
Oncogene Proteins
Sample Size
Complementary DNA
RNA
Staining and Labeling
Cell Line
Genes
Neoplasms

ASJC Scopus subject areas

  • Pathology and Forensic Medicine

Cite this

Quantitative analysis of Her-2/neu (ERBB2) gene expression using reverse transcriptase polymerase chain reaction. / Sarkar, F. H.; Visscher, Daniel W; Crissman, J. D.

In: Diagnostic Molecular Pathology, Vol. 2, No. 3, 1993, p. 210-218.

Research output: Contribution to journalArticle

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