title: In vitro characterisation of the anti-intravasative properties of the marine product heteronemin.
abstract: Metastases destroy the function of infested organs and are the main reason of cancer-related mortality. Heteronemin, a natural product derived from a marine sponge, was tested in vitro regarding its properties to prevent tumour cell intravasation through the lymph-endothelial barrier. In three-dimensional (3D) cell cultures consisting of MCF-7 breast cancer cell spheroids that were placed on lymph-endothelial cell (LEC) monolayers, tumour cell spheroids induce "circular chemorepellent-induced defects" (CCIDs) in the LEC monolayer; 12(S)-Hydroxyeicosatetraenoic acid (12(S)-HETE) and NF-κB activity are major factors inducing CCIDs, which are entry gates for tumour emboli intravasating the vasculature. This 3D co-culture is a validated model for the investigation of intravasation mechanisms and of drugs preventing CCID formation and hence lymph node metastasis. Furthermore, Western blot analyses, NF-κB reporter, EROD, SELE, 12(S)-HETE, and adhesion assays were performed to investigate the properties of heteronemin. Five micromolar heteronemin inhibited the directional movement of LECs and, therefore, the formation of CCIDs, which were induced by MCF-7 spheroids. Furthermore, heteronemin reduced the adhesion of MCF-7 cells to LECs and suppressed 12(S)-HETE-induced expression of the EMT marker paxillin, which is a regulator of directional cell migration. The activity of CYP1A1, which contributed to CCID formation, was also inhibited by heteronemin. Hence, heteronemin inhibits important mechanisms contributing to tumour intravasation in vitro and should be tested in vivo.
authors.affiliation: Institute of Clinical Pathology, Medical University of Vienna, Waehringer Guertel 18-20, 1090, Vienna, Austria.
authors.forename: Sabine
authors.initials: S
authors.lastname: Kopf
authors.forename: Katharina
authors.initials: K
authors.lastname: Viola
authors.forename: Atanas G
authors.initials: AG
authors.lastname: Atanasov
authors.forename: Kanokwan
authors.initials: K
authors.lastname: Jarukamjorn
authors.forename: Lucie
authors.initials: L
authors.lastname: Rarova
authors.forename: Nicole
authors.initials: N
authors.lastname: Kretschy
authors.forename: Mathias
authors.initials: M
authors.lastname: Teichmann
authors.forename: Caroline
authors.initials: C
authors.lastname: Vonach
authors.forename: Philipp
authors.initials: P
authors.lastname: Saiko
authors.forename: Benedikt
authors.initials: B
authors.lastname: Giessrigl
authors.forename: Nicole
authors.initials: N
authors.lastname: Huttary
authors.forename: Ingrid
authors.initials: I
authors.lastname: Raab
authors.forename: Sigurd
authors.initials: S
authors.lastname: Krieger
authors.forename: Marc
authors.initials: M
authors.lastname: Schumacher
authors.forename: Marc
authors.initials: M
authors.lastname: Diederich
authors.forename: Miroslav
authors.initials: M
authors.lastname: Strnad
authors.forename: Rainer
authors.initials: R
authors.lastname: de Martin
authors.forename: Thomas
authors.initials: T
authors.lastname: Szekeres
authors.forename: Walter
authors.initials: W
authors.lastname: Jäger
authors.forename: Verena M
authors.initials: VM
authors.lastname: Dirsch
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Mikulits
authors.forename: Michael
authors.initials: M
authors.lastname: Grusch
authors.forename: Helmut
authors.initials: H
authors.lastname: Dolznig
authors.forename: Georg
authors.initials: G
authors.lastname: Krupitza
mesh_terms: D019377:12-Hydroxy-5,8,10,14-eicosatetraenoic Acid / Q000378:metabolism; D015153:Blotting, Western; D001943:Breast Neoplasms / Q000188:drug therapy* / Q000473:pathology; D002465:Cell Movement; D018920:Coculture Techniques; D019363:Cytochrome P-450 CYP1A1 / Q000378:metabolism; D042783:Endothelial Cells / Q000187:drug effects* / Q000378:metabolism; D005260:Female; D006801:Humans; D008207:Lymphatic Metastasis / Q000517:prevention & control*; D061986:MCF-7 Cells; D016328:NF-kappa B / Q000378:metabolism; D051419:Paxillin / Q000378:metabolism; D013729:Terpenes / Q000494:pharmacology*
chemical_list: D016328:NF-kappa B; D051419:Paxillin; D013729:Terpenes; C493748:heteronemin; D019377:12-Hydroxy-5,8,10,14-eicosatetraenoic Acid; D019363:Cytochrome P-450 CYP1A1