title: NF-κB mediates the 12(S)-HETE-induced endothelial to mesenchymal transition of lymphendothelial cells during the intravasation of breast carcinoma cells.
abstract: BACKGROUND
  The intravasation of breast cancer into the lymphendothelium is an early step of metastasis. Little is known about the mechanisms of bulky cancer invasion into lymph ducts.
  
  
  METHODS
  To particularly address this issue, we developed a 3-dimensional co-culture model involving MCF-7 breast cancer cell spheroids and telomerase-immortalised human lymphendothelial cell (LEC) monolayers, which resembles intravasation in vivo and correlated the malignant phenotype with specific protein expression of LECs.
  
  
  RESULTS
  We show that tumour spheroids generate 'circular chemorepellent-induced defects' (CCID) in LEC monolayers through retraction of LECs, which was induced by 12(S)-hydroxyeicosatetraenoic acid (HETE) secreted by MCF-7 spheroids. This 12(S)-HETE-regulated retraction of LECs during intravasation particularly allowed us to investigate the key regulators involved in the motility and plasticity of LECs. In all, 12(S)-HETE induced pro-metastatic protein expression patterns and showed NF-κB-dependent up-regulation of the mesenchymal marker protein S100A4 and of transcriptional repressor ZEB1 concomittant with down-regulation of the endothelial adherence junction component VE-cadherin. This was in accordance with ∼50% attenuation of CCID formation by treatment of cells with 10 μM Bay11-7082. Notably, 12(S)-HETE-induced VE-cadherin repression was regulated by either NF-κB or by ZEB1 since ZEB1 siRNA knockdown abrogated not only 12(S)-HETE-mediated VE-cadherin repression but inhibited VE-cadherin expression in general.
  
  
  CONCLUSIONS
  These data suggest an endothelial to mesenchymal transition-like process of LECs, which induces single cell motility during endothelial transmigration of breast carcinoma cells. In conclusion, this study demonstrates that the 12(S)-HETE-induced intravasation of MCF-7 spheroids through LECs require an NF-κB-dependent process of LECs triggering the disintegration of cell-cell contacts, migration, and the generation of CCID.
authors.affiliation: Institute of Clinical Pathology, Medical University of Vienna, Waehringer Guertel 18-20, A-1090 Vienna, Austria.
authors.forename: C
authors.initials: C
authors.lastname: Vonach
authors.forename: K
authors.initials: K
authors.lastname: Viola
authors.forename: B
authors.initials: B
authors.lastname: Giessrigl
authors.forename: N
authors.initials: N
authors.lastname: Huttary
authors.forename: I
authors.initials: I
authors.lastname: Raab
authors.forename: R
authors.initials: R
authors.lastname: Kalt
authors.forename: S
authors.initials: S
authors.lastname: Krieger
authors.forename: T P N
authors.initials: TP
authors.lastname: Vo
authors.forename: S
authors.initials: S
authors.lastname: Madlener
authors.forename: S
authors.initials: S
authors.lastname: Bauer
authors.forename: B
authors.initials: B
authors.lastname: Marian
authors.forename: M
authors.initials: M
authors.lastname: Hämmerle
authors.forename: N
authors.initials: N
authors.lastname: Kretschy
authors.forename: M
authors.initials: M
authors.lastname: Teichmann
authors.forename: B
authors.initials: B
authors.lastname: Hantusch
authors.forename: S
authors.initials: S
authors.lastname: Stary
authors.forename: C
authors.initials: C
authors.lastname: Unger
authors.forename: M
authors.initials: M
authors.lastname: Seelinger
authors.forename: A
authors.initials: A
authors.lastname: Eger
authors.forename: R
authors.initials: R
authors.lastname: Mader
authors.forename: W
authors.initials: W
authors.lastname: Jäger
authors.forename: W
authors.initials: W
authors.lastname: Schmidt
authors.forename: M
authors.initials: M
authors.lastname: Grusch
authors.forename: H
authors.initials: H
authors.lastname: Dolznig
authors.forename: W
authors.initials: W
authors.lastname: Mikulits
authors.forename: G
authors.initials: G
authors.lastname: Krupitza
mesh_terms: D019377:12-Hydroxy-5,8,10,14-eicosatetraenoic Acid / Q000494:pharmacology*; D000970:Antineoplastic Agents / Q000494:pharmacology; D001943:Breast Neoplasms / Q000378:metabolism / Q000473:pathology*; D002277:Carcinoma / Q000378:metabolism / Q000473:pathology*; D002461:Cell Line, Transformed; D002465:Cell Movement / Q000187:drug effects; D054338:Cell Transdifferentiation / Q000187:drug effects*; D018920:Coculture Techniques; D042783:Endothelial Cells / Q000187:drug effects* / Q000502:physiology; D005260:Female; D006801:Humans; D008648:Mesoderm / Q000187:drug effects / Q000502:physiology; D016328:NF-kappa B / Q000037:antagonists & inhibitors / Q000378:metabolism / Q000502:physiology*; D009361:Neoplasm Invasiveness; D009570:Nitriles / Q000494:pharmacology; D015398:Signal Transduction / Q000187:drug effects; D013450:Sulfones / Q000494:pharmacology; D014407:Tumor Cells, Cultured
chemical_list: C434003:3-(4-methylphenylsulfonyl)-2-propenenitrile; D000970:Antineoplastic Agents; D016328:NF-kappa B; D009570:Nitriles; D013450:Sulfones; D019377:12-Hydroxy-5,8,10,14-eicosatetraenoic Acid