title: Real-time quantitative reverse transcriptase-polymerase chain reaction for luteinizing hormone-releasing hormone receptor gene mRNA expression in human prostate cancer.
abstract: OBJECTIVE
  To determine quantitative real-time reverse transcriptase-polymerase chain reaction (RT-PCR) detection of luteinizing hormone-releasing hormone (LHRH) receptor mRNA expressing prostate cancer (CaP) and benign prostatic hyperplasia (BPH) tissue by sequence-specific detection with hybridization probes using the LightCycler. Various in vitro studies have indicated that LHRH agonists and antagonists may have a direct inhibitory effect on CaP mediated by specific LHRH receptors. The incidence of LHRH receptor mRNA expression was demonstrated in 50% to 100% of clinical CaP and BPH tissue by current RT-PCR methods. However, qualitative RT-PCR can only demonstrate the existence of a gene; an exact quantification of LHRH-receptor mRNA expression has not yet been done.
  
  
  METHODS
  Quantitative real-time RT-PCR for LHRH receptor mRNA was performed using the LightCycler system and the RNA Amplification Kit hybridization probes in 35 patients with CaP and 38 patients with BPH.
  
  
  RESULTS
  Thirty-one (88.6%) of the 35 patients with CaP and 36 (94.7%) of the 38 patients with BPH had positive RT-PCR for LHRH receptor mRNA. The number of positive amplifications per 1 microg total RNA was not significantly lower in CaP at 695,428 +/- 350,860 than in BPH at 1,617,654 +/- 787,874. Patients with CaP evidenced a significant negative correlation between the amplification rate for LHRH receptor mRNA and Gleason score (r = -0.476; P = 0.004).
  
  
  CONCLUSIONS
  In the past, the potential mechanisms acting on LHRH receptors in CaP have been identified as possible antitumor strategies for this type of cancer. Our study is the first to show that CaP does not have a higher expression of LHRH receptor mRNA expression than BPH when using quantitative sequence-specific oligonucleotide hybridization probes that only detect certain PCR products.
authors.affiliation: Department of Urology, Universitätsklinikum Benjamin Franklin, Freie Universität Berlin, Berlin, Germany.
authors.forename: Bernd
authors.initials: B
authors.lastname: Straub
authors.forename: Markus
authors.initials: M
authors.lastname: Müller
authors.forename: Hans
authors.initials: H
authors.lastname: Krause
authors.forename: Mark
authors.initials: M
authors.lastname: Schrader
authors.forename: Kurt
authors.initials: K
authors.lastname: Miller
mesh_terms: D000230:Adenocarcinoma / Q000235:genetics* / Q000378:metabolism; D000368:Aged; D018931:Antineoplastic Agents, Hormonal / Q000494:pharmacology; D003199:Computer Systems; D015972:Gene Expression Regulation, Neoplastic; D007987:Gonadotropin-Releasing Hormone / Q000819:agonists; D006801:Humans; D008297:Male; D008875:Middle Aged; D009363:Neoplasm Proteins / Q000096:biosynthesis* / Q000235:genetics; D009376:Neoplasms, Hormone-Dependent / Q000235:genetics* / Q000378:metabolism; D011470:Prostatic Hyperplasia / Q000235:genetics / Q000378:metabolism; D011471:Prostatic Neoplasms / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000096:biosynthesis* / Q000235:genetics; D012334:RNA, Neoplasm / Q000096:biosynthesis* / Q000235:genetics; D011966:Receptors, LHRH / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D013739:Testosterone
chemical_list: D018931:Antineoplastic Agents, Hormonal; D009363:Neoplasm Proteins; D012333:RNA, Messenger; D012334:RNA, Neoplasm; D011966:Receptors, LHRH; D007987:Gonadotropin-Releasing Hormone; D013739:Testosterone