title: Secreted ovarian stromal substance inhibits ovarian epithelial cell proliferation.
abstract: OBJECTIVE
  Determine the effects of factors secreted by normal human ovarian stroma on the proliferation of benign and malignant ovarian epithelia, in vitro.
  
  
  METHODS
  Primary cultures of normal human ovarian surface epithelium (HOSE), human ovarian stromal tissue (HOST), and epithelial ovarian carcinomas (CSOC) were established from surgical specimens and characterized immunohistochemically using anti-cytokeratin, vimentin, and Factor VIII antibodies. Stroma-conditioned media (SCM) were collected over 3 days from confluent HOST cultures. The SCM were dialyzed, lyophilized, resuspended, and added to HOSE, CSOC, SKOV-3, and Caov-3 ovarian cancer cell cultures and growth inhibitory effects were assayed by MTS and [3H]thymidine uptake.
  
  
  RESULTS
  SCM inhibited the growth and DNA synthesis of normal HOSE cells and cancer cells by 79-99% in > 10-cell lines studied to date. The inhibitory effect was rapid in onset with 31-82% reduction in DNA synthesis at 1 hr and approximately 50% return of activity by 23 hr following a 1-hr SCM pulse treatment. The SCM inhibitory activity was not abolished by boiling or by absorption with heparin-agarose. Size exclusion filtration places the molecular weight of the inhibitory substance between 1 and 3 kDa. Neither trypsin nor proteinase K treatments altered the inhibitory activity of SCM, while a Bligh-Dyer organic extraction placed the activity in the aqueous phase.
  
  
  CONCLUSIONS
  A heat-stable, non-heparin-binding, low-molecular-weight, water-soluble substance secreted by normal ovarian stroma significantly inhibits HOSE and ovarian cancer cell proliferation. Derangements in normal ovarian stroma-epithelial interactions may contribute to growth dysregulation of the surface epithelia and result in ovarian carcinogenesis.
authors.affiliation: Department of Obstetrics and Gynecology, Cedars-Sinai Medical Center, University of California, Los Angeles, School of Medicine 90048, USA.
authors.forename: B Y
authors.initials: BY
authors.lastname: Karlan
authors.forename: R L
authors.initials: RL
authors.lastname: Baldwin
authors.forename: F D
authors.initials: FD
authors.lastname: Cirisano
authors.forename: P W
authors.initials: PW
authors.lastname: Mamula
authors.forename: J
authors.initials: J
authors.lastname: Jones
authors.forename: L D
authors.initials: LD
authors.lastname: Lagasse
mesh_terms: D001685:Biological Factors / Q000378:metabolism / Q000502:physiology*; D002455:Cell Division; D002478:Cells, Cultured; D017077:Culture Media, Conditioned; D004847:Epithelial Cells; D004848:Epithelium / Q000378:metabolism; D005260:Female; D006801:Humans; D008970:Molecular Weight; D010051:Ovarian Neoplasms / Q000473:pathology*; D010053:Ovary / Q000166:cytology* / Q000378:metabolism*; D017154:Stromal Cells / Q000378:metabolism
chemical_list: D001685:Biological Factors; D017077:Culture Media, Conditioned