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 progr ession of breast cancer. In order to refine and facilitate the quantit ative analysis of HER-2 amplification in breast cancer, differential p olymerase chain reaction (PCR) was performed on DNA derived from singl e cryosections of tumor tissue. This technique is based on the simulta neous amplification of a potentially amplified oncogene (HER-2) and a reference gene (IFN-gamma). Differential PCR yielded reproducible resu lts 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.