LNCaP produces both putative zymogen and inactive, free form of prostate-specific antigen

Prostate. 1998 May;35(2):135-43. doi: 10.1002/(sici)1097-0045(19980501)35:2<135::aid-pros7>3.0.co;2-i.

Abstract

Background: Our objective was to investigate the presence of prostate specific antigen (PSA) and alpha-1-antichymotrypsin (ACT) mRNA and protein in prostate cancer cell lines, and the complexing characteristics of expressed PSA.

Methods: RT-PCR, immunohistochemistry, and Western blots were employed. Trypsin treatment of PSA was performed to establish the possible presence of an activatable form of PSA.

Results: ACT mRNA and protein were detected in LNCaP, PC-3, and DU 145 by RT-PCR and by immunohistochemistry, respectively. Only LNCaP cells were positive for PSA mRNA and protein. LNCaP expressed approximately 30% active PSA, approximately 40% putative zymogen form of PSA, and approximately 30% stably inactive PSA.

Conclusions: We have shown that the majority of PSA expressed by LNCaP cells is present in free, noncomplexed forms in the conditioned media. A portion (40%) can be activated by trypsin, while the rest is stably inactive PSA. LNCaP cells may serve as a source of the "unreactive" PSA present in prostate cancer patients' serum.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biomarkers, Tumor / analysis*
  • Blotting, Western
  • DNA Primers
  • Enzyme Precursors / analysis*
  • Humans
  • Immunohistochemistry
  • Male
  • Peptides / analysis*
  • Polymerase Chain Reaction / methods
  • Prostate-Specific Antigen / analysis*
  • Prostate-Specific Antigen / genetics
  • Prostatic Neoplasms / enzymology*
  • Prostatic Neoplasms / immunology*
  • Prostatic Secretory Proteins*
  • RNA, Messenger / analysis
  • RNA, Neoplasm / analysis
  • RNA-Directed DNA Polymerase
  • Tumor Cells, Cultured
  • alpha 1-Antichymotrypsin / analysis*
  • alpha 1-Antichymotrypsin / genetics

Substances

  • Biomarkers, Tumor
  • DNA Primers
  • Enzyme Precursors
  • Peptides
  • Prostatic Secretory Proteins
  • RNA, Messenger
  • RNA, Neoplasm
  • alpha 1-Antichymotrypsin
  • beta-microseminoprotein
  • RNA-Directed DNA Polymerase
  • Prostate-Specific Antigen