14-3-3 Zeta protein secreted by tumor associated monocytes/macrophages from ascites of epithelial ovarian cancer patients

Ryuji Kobayashi, Michael Deavers, Rebecca Patenia, Travis Rice-Stitt, Jeremy Halbe, Stacie Gallardo, Ralph S. Freedman

Research output: Contribution to journalArticlepeer-review

37 Scopus citations

Abstract

Tumor associated monocytes/macrophages (MO/MA) are known contributors to the immune-inflammatory cell environment of advanced epithelial ovarian carcinoma (EOC). The secreted proteome of ascitic MO/ MA was examined as an aid to the discovery of novel proteins in EOC that are likely to have biological relevance in the inflammatory pathways of EOC. Ascitic fluid MO/ MA were isolated from EOC patients, grown short-term in serum-free media. MO/MA supernatants were analyzed for secreted proteins by HPLC fractionation followed by LC-tandem mass spectrometric analysis. The 14-3-3 zeta adaptor protein was identified in supernatants of three of three EOC patients but not in supernatants of buffy coat monocytes isolated from normal donors or the established monocyte cell line THP1. Moreover, 14-3-3 zeta was identified in ascitic fluids in eight of eight chemotherapy naïve patients by both immunoblot and mass spectrometric analysis. Immunofluorescent staining for 14-3-3 zeta demonstrated expression of the protein on ascitic and peritumoral macrophages in EOC patients. 14-3-3 zeta was also expressed on endothelial cells in the peritumoral stroma and partially on tumor cells. Uptake of 14-3-3 zeta was observed in EOC cell lines co-cultured with the recombinant protein expressed in E. coli. It is demonstrated for the first time that the important adaptor protein 14-3-3 zeta is common to the secretome of ascitic MO/MA and the ascites of advanced EOC patients.

Original languageEnglish (US)
Pages (from-to)247-258
Number of pages12
JournalCancer Immunology, Immunotherapy
Volume58
Issue number2
DOIs
StatePublished - Feb 2009

Keywords

  • Inflammation
  • Monocytes/Macrophages
  • Ovarian cancer
  • Secreted proteins 14-3-3 zeta

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology
  • Oncology
  • Cancer Research

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