Detection of HER-2 oncogene amplification in breast cancer by differential polymerase chain reaction from single cryosections.
Amplification of genomic DNA encoding oncogenes such as HER-2 (syn.c-erbB2/c-neu) may be substantially involved in the initiation and progression of breast cancer. In order to refine and facilitate the quantitative analysis of HER-2 amplification in breast cancer, differential polymerase chain reaction (PCR) was performed on DNA derived from single cryosections of tumor tissue. This technique is based on the simultaneous amplification of a potentially amplified oncogene (HER-2) and a reference gene (IFN-gamma). Differential PCR yielded reproducible results that were in agreement with gene copy quantification using the dot blot technique. Thus we suggest differential PCR to be a reliable and rapid method for determining relative gene dosage in a minute amount of tumour tissue.
Friedrichs
K
K
Department of Gynecology and Obstetrics, University of Hamburg, Medical School, Germany.
Lohmann
D
D
Höfler
H
H
D001483:Base Sequence; D001943:Breast Neoplasms / Q000235:genetics*; D004273:DNA, Neoplasm / Q000302:isolation & purification; D066246:ErbB Receptors / Q000235:genetics*; D005260:Female; D005629:Frozen Sections; D005784:Gene Amplification / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D009857:Oncogenes / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011518:Proto-Oncogene Proteins / Q000235:genetics*; D018719:Erb-b2 Receptor Tyrosine Kinases
D004273:DNA, Neoplasm; D066246:ErbB Receptors; D011518:Proto-Oncogene Proteins; D018719:Erb-b2 Receptor Tyrosine Kinases