title: Cancer cells cause vascular endothelial cell (vEC) retraction via 12(S)HETE secretion; the possible role of cancer cell derived microparticle.
abstract: BACKGROUND
  Cancer cell mediated vascular endothelial cell (vEC) retraction plays a pivotal role in cancer metastasis. The aim of this study is to clarify the biochemical character of vEC retraction factor derived from human breast cancer cell line, MCF-7.
  
  
  RESULTS
  In order to estimate vEC retracting activity, transwell chamber assay system was employed. We first tested the effects of trypsin digestion as well as lipid extraction of culture medium (CM). Trypsin digestion of CM resulted in approximately 40% loss of vEC retracting activity and lipid extraction of CM by Brigh and Dyer methods recovered approximately 60% of vEC retracting activity, suggesting that approximately 60% of vEC retracting activity in MCF-7 derived CM is due to lipid. Although Nordihydroguaiaretic acid (NDGA), the specific lipoxygenase inhibitor, suppressed vEC retracting activity in CM, Acetyl salicylic acid (ASA), a specific cyclooxygenase inhibitor, did not affect the activity, suggesting that lipid exerting vEC retracting activity in CM belongs to lipoxygenase mediated arachidonate metabolites. Thin layer chromatography clearly demonstrated that Rf value of lipid vEC retracting factor in CM is identical to 12HETE. Authentic 12(S)HETE, but not 12(R)HETE, showed vEC retracting activity. After the ultracentrifugation of CM, most lipid vEC retracting activity was recovered from the pellet fraction, and flow cytometric analysis using specific antibody against 12(S)HETE clearly showed the association of 12(S)HETE with small particle in CM.
  
  
  CONCLUSIONS
  These findings suggested the principal involvement of 12(S)HETE in cancer cell derived microparticles in cancer cell mediated vEC retraction.
authors.affiliation: Department of Gastroenterological Surgery, Graduate School of Medicine, Osaka University, 2-2, Yamadaoka, Suitacity, Osaka, Japan. k-uchide@jb3.so-net.ne.jp
authors.forename: Keiji
authors.initials: K
authors.lastname: Uchide
authors.forename: Masato
authors.initials: M
authors.lastname: Sakon
authors.forename: Hideo
authors.initials: H
authors.lastname: Ariyoshi
authors.forename: Syouji
authors.initials: S
authors.lastname: Nakamori
authors.forename: Masaru
authors.initials: M
authors.lastname: Tokunaga
authors.forename: Morito
authors.initials: M
authors.lastname: Monden
mesh_terms: D019377:12-Hydroxy-5,8,10,14-eicosatetraenoic Acid / Q000378:metabolism*; D001943:Breast Neoplasms / Q000378:metabolism*; D002448:Cell Adhesion / Q000502:physiology; D045744:Cell Line, Tumor; D042783:Endothelial Cells / Q000378:metabolism*; D005260:Female; D006801:Humans; D009362:Neoplasm Metastasis / Q000503:physiopathology*
chemical_list: D019377:12-Hydroxy-5,8,10,14-eicosatetraenoic Acid