title: Reverse transcription-polymerase chain reaction detection of prostate-specific antigen, prostate-specific membrane antigen, and prostate stem cell antigen in one milliliter of peripheral blood: value for the staging of prostate cancer.
abstract: OBJECTIVE
  There have been several studies on the presence of circulating tumor cells in the peripheral blood of patients with malignant tumors including prostate cancer (PCa) using reverse transcription-PCR (RT-PCR). One of the aims of these studies was to obtain high sensitivity that would enable early-stage diagnosis. However, they varied in their detection rates, and the methods were rather complicated. We have improved the RT-PCR assay combining three prostate-associated molecules, prostate specific antigen (PSA), prostate specific membrane antigen (PSMA), and prostate stem cell antigen (PSCA) to reveal patients with poor prognosis.
  
  
  METHODS
  Peripheral blood samples were obtained from 129 patients including 58 cases of PCa and 71 cases of nonmalignant disorders. Total RNA was extracted from 1 ml of whole blood using a commercially available kit.
  
  
  RESULTS
  The sensitivity of PSA-, PSMA-, and PSCA-nested RT-PCR was verified with positive signals of a single LNCaP cell in 1 ml of female blood sample. PSA-, PSMA-, and PSCA-mRNA were detected in 7 (12.1%), 12 (20.7%), and 8 (13.8%) PCa, and in 1, 2, and 0 samples in nonmalignant disorders, respectively. Among 58 PCa patients, each PCR indicated the prognostic value in the hierarchy of PSCA>PSA>PSMA RT-PCR, and extraprostatic cases with positive PSCA PCR indicated lower disease-progression-free survival than those with negative PSCA PCR.
  
  
  CONCLUSIONS
  The present findings suggest that PSCA PCR would be most promising for the molecular staging of PCa. The present RT-PCR is a highly cost-effective and rapid procedure, enabling the molecular staging of PCa with RT-PCR as a diagnostic routine.
authors.affiliation: Division of Molecular Oncology, Department of Signal Transduction Research, Graduate School of Medical and Dental Sciences, Niigata University, Asahimachi 1, Niigata 951, Japan. harasho@ med.niigata-u.ac.jp
authors.forename: Noboru
authors.initials: N
authors.lastname: Hara
authors.forename: Takashi
authors.initials: T
authors.lastname: Kasahara
authors.forename: Takashi
authors.initials: T
authors.lastname: Kawasaki
authors.forename: Vladimir
authors.initials: V
authors.lastname: Bilim
authors.forename: Kenji
authors.initials: K
authors.lastname: Obara
authors.forename: Kota
authors.initials: K
authors.lastname: Takahashi
authors.forename: Yoshihiko
authors.initials: Y
authors.lastname: Tomita
mesh_terms: D000951:Antigens, Neoplasm; D014408:Biomarkers, Tumor / Q000032:analysis*; D017931:DNA Primers / Q000737:chemistry; D018450:Disease Progression; D018572:Disease-Free Survival; D058851:GPI-Linked Proteins; D006801:Humans; D008297:Male; D008562:Membrane Glycoproteins / Q000097:blood*; D009363:Neoplasm Proteins / Q000097:blood*; D009367:Neoplasm Staging; D011446:Prospective Studies; D017430:Prostate-Specific Antigen / Q000097:blood*; D011470:Prostatic Hyperplasia / Q000097:blood / Q000473:pathology; D019048:Prostatic Intraepithelial Neoplasia / Q000097:blood / Q000473:pathology; D011471:Prostatic Neoplasms / Q000097:blood / Q000473:pathology*; D012334:RNA, Neoplasm / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D013997:Time Factors
chemical_list: D000951:Antigens, Neoplasm; D014408:Biomarkers, Tumor; D017931:DNA Primers; D058851:GPI-Linked Proteins; D008562:Membrane Glycoproteins; D009363:Neoplasm Proteins; C110843:PSCA protein, human; D012334:RNA, Neoplasm; D017430:Prostate-Specific Antigen