title: Expression of prostatic factors measured by reverse transcription polymerase chain reaction in human papillomavirus type 18 deoxyribonucleic acid immortalized prostate cell lines.
abstract: OBJECTIVE
  To investigate expression of the prostatic markers prostate-specific antigen (PSA), prostate-specific membrane antigen (PSM), and the androgen receptor (AR) after human papillomavirus (HPV) type 18 deoxyribonucleic acid (DNA) transfection and subsequent immortalization of human prostate epithelial cells.
  
  
  METHODS
  Recently, two human prostate epithelial cell lines were established by HPV transformation: PZ-HPV-7, derived from normal peripheral zone (PZ) tissue, and CA-HPV-10, derived from high Gleason grade adenocarcinoma. Expression of PSA was studied by the reverse transcription polymerase chain reaction (RT-PCR), because in preliminary studies using immunocytochemistry and Northern blotting, no PSA expression was found. PSM was analyzed by RT-PCR and nested RT-PCR. These analyses included primary human prostate cell strains. Furthermore, androgen-supplemented methylthiazol tetrazolium (MTT) growth assays were performed and expression of AR was studied by immunocytochemistry. Prostate carcinoma cell lines LNCaP and PC-346C were included as positive controls and breast carcinoma cell line MCF-7 as a negative control.
  
  
  RESULTS
  Both cell lines exhibited low levels of RNA for PSA and PSM in comparison with cell lines LNCaP and PC-346C. AR expression by immunocytochemistry was negative using monoclonal antibody F39.4 and polyclonal antibody SP-197. In an androgen-supplemented environment, growth rates of both HPV immortalized cell lines were not stimulated in contrast to LNCaP.
  
  
  CONCLUSIONS
  RNA transcripts of PSA and PSM were detected by RT-PCR in HPV immortalized prostate epithelial cell lines PZ-HPV-7 and CA-HPV-10. The expression of prostate-specific markers may further validate the utility of this stepwise transformation model of human prostate carcinogenesis.
authors.affiliation: Department of Urology, Erasmus University Rotterdam, The Netherlands.
authors.forename: P C
authors.initials: PC
authors.lastname: Weijerman
authors.forename: Y
authors.initials: Y
authors.lastname: Zhang
authors.forename: J
authors.initials: J
authors.lastname: Shen
authors.forename: H J
authors.initials: HJ
authors.lastname: Dubbink
authors.forename: J C
authors.initials: JC
authors.lastname: Romijn
authors.forename: D M
authors.initials: DM
authors.lastname: Peehl
authors.forename: F H
authors.initials: FH
authors.lastname: Schröder
mesh_terms: D000954:Antigens, Surface / Q000032:analysis / Q000096:biosynthesis*; D002268:Carboxypeptidases / Q000032:analysis / Q000096:biosynthesis*; D002478:Cells, Cultured; D043425:Glutamate Carboxypeptidase II; D006801:Humans; D008297:Male; D027383:Papillomaviridae / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011467:Prostate / Q000166:cytology / Q000378:metabolism*; D017430:Prostate-Specific Antigen / Q000032:analysis / Q000096:biosynthesis*; D011944:Receptors, Androgen / Q000032:analysis / Q000096:biosynthesis*; D014162:Transfection
chemical_list: D000954:Antigens, Surface; D011944:Receptors, Androgen; D002268:Carboxypeptidases; C473848:FOLH1 protein, human; D043425:Glutamate Carboxypeptidase II; D017430:Prostate-Specific Antigen