Gene Amplification and Overexpression in BPH by Means of One-step Real Time Quantitative PCR / 대한남성과학회지

ABSTRACT

PURPOSE: Gene amplification/overexpression of c-erbB2/neu is controversy in BPH by conventional detecting methods. To evaluate the amplification and overexpression of c-erbB2/neu gene in BPH we have used and validated a one-step real time PCR and quantitative reverse transcription PCR assay based on fluorescent TaqMan methodology. MATERIALS AND METHODS: Using one-step real time PCR assay, we have assesed the amplification and overexpression of c-erbB2/neu gene in 30 prostate samples from patients with BPH as well as from 10 normal blood sample. Real time PCR and RT-PCR were performed on an iCycler (Bio-Rad Co., USA) and we used TaqMan probes to detect c-erbB2/neu transcripts amplified. All experiments were performed in duplicate using 96 well-PCR plate. The GAPDH housekeeping gene was used for normalization of c-erbB2/neu amplification and overexpression. RESULTS: In a series of 30 BPH samples c-erbB2/neu normalized amplification was found to range from 3.16 10(-3) to 7.16 10(-2) and overexpression to range from 7.72 10(-3) to 1.57 10. Sixteen cases of BPH (53.3%) showed c-erbB2/neu overexpression and only two cases (6.7%) showed amplification of c-erbB2/neu. Except 2 cases, no correlation was found between the results of amplification and overexpression of c-erbB2/neu (p=0.183). CONCLUSIONS: Our results using of one-step real time quantitative PCR assay suggest that a role of c-erbB2/neu overexpression in the development of BPH and the use of this new semi-automated technique will make molecular analysis of gene simpler and more reliable.