Detection of Amplified Oncogenes by Differential Polymerase Chain Reaction
Genetic alterations such as oncogene amplifications have been found in a variety of human cancers and may have a prognostic impact. Therefore we used a non-radioactive tool for possible routine screening of genetic alterations with high fidelity in human cancer probes. This technique for detecting oncogene amplifications is a new application of the recently published method by Frye et al., called differential polymerase chain reaction (PCR). With this PCR we amplified five oncogenes which are involved in breast tumor metabolism (erb B-1, erb B-2, erb B-3, c-myc and Ha-ras). The level of amplification is reflected by the ratio of the oncogene and the single-copy reference genes (ß-globin, phenylalanine-hydroxylase) in gel electrophoresis. In a study of 169 breast cancer tissues, we showed that differential PCR is able to detect at least three fold oncogene amplifications. We regarded an oncogene as amplified if the cut off level of three fold amplification was passed. In about 40% of the carcinoma tissues we detected gene amplifications of at least one oncogene.
KeywordsPhenylalanine Hydroxylase Oncogene Amplification Healthy Human Donor Fold Amplification Polymerase Chain Reaction Signal
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