Archives of Toxicology

, Volume 87, Issue 10, pp 1851–1861 | Cite as

In vitro characterisation of the anti-intravasative properties of the marine product heteronemin

  • Sabine Kopf
  • Katharina Viola
  • Atanas G. Atanasov
  • Kanokwan Jarukamjorn
  • Lucie Rarova
  • Nicole Kretschy
  • Mathias Teichmann
  • Caroline Vonach
  • Philipp Saiko
  • Benedikt Giessrigl
  • Nicole Huttary
  • Ingrid Raab
  • Sigurd Krieger
  • Marc Schumacher
  • Marc Diederich
  • Miroslav Strnad
  • Rainer de Martin
  • Thomas Szekeres
  • Walter Jäger
  • Verena M. Dirsch
  • Wolfgang Mikulits
  • Michael Grusch
  • Helmut Dolznig
  • Georg Krupitza
Biologicals

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.

Keywords

Heteronemin Intravasation Lymph endothelium Adhesion Mobility 

Abbreviations

ALOX

Lipoxygenase A

CCIDs

Circular chemorepellent-induced defects

CYP

Cytochrome P450

EMT

Epithelial to mesenchymal transition

EROD

Ethoxyresorufin-O-deethylase

HUVEC

Human umbilical vein endothelial cell

ICAM-1

Intercellular adhesion molecule 1

LEC

Lymph-endothelial cell

MCL2

Myosin light chain 2

SELE

Selectin E

12(S)-HETE 12(S)

Hydroxyeicosatetraenoic acid

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Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • Sabine Kopf
    • 1
  • Katharina Viola
    • 1
  • Atanas G. Atanasov
    • 2
  • Kanokwan Jarukamjorn
    • 3
    • 4
  • Lucie Rarova
    • 5
  • Nicole Kretschy
    • 1
  • Mathias Teichmann
    • 1
  • Caroline Vonach
    • 1
  • Philipp Saiko
    • 6
  • Benedikt Giessrigl
    • 1
  • Nicole Huttary
    • 1
  • Ingrid Raab
    • 1
  • Sigurd Krieger
    • 1
  • Marc Schumacher
    • 7
  • Marc Diederich
    • 7
  • Miroslav Strnad
    • 5
  • Rainer de Martin
    • 8
  • Thomas Szekeres
    • 6
  • Walter Jäger
    • 3
  • Verena M. Dirsch
    • 2
  • Wolfgang Mikulits
    • 9
  • Michael Grusch
    • 9
  • Helmut Dolznig
    • 10
  • Georg Krupitza
    • 1
  1. 1.Institute of Clinical PathologyMedical University of ViennaViennaAustria
  2. 2.Department of PharmacognosyUniversity of ViennaViennaAustria
  3. 3.Department for Clinical Pharmacy and Diagnostics, Faculty of Life SciencesUniversity of ViennaViennaAustria
  4. 4.Department of Pharmaceutical Chemistry, Faculty of Pharmaceutical SciencesKhon Kaen UniversityKhon KaenThailand
  5. 5.Faculty of Science, Centre of the Region Haná for Biotechnological and Agricultural ResearchPalacký UniversityOlomoucCzech Republic
  6. 6.Department of Medical and Chemical Laboratory Diagnostics, General Hospital of ViennaMedical University of ViennaViennaAustria
  7. 7.Laboratoire de Biologie Moléculaire et Cellulaire du CancerHôpital KirchbergLuxembourgLuxembourg
  8. 8.Department of Vascular Biology and Thrombosis ResearchMedical University of ViennaViennaAustria
  9. 9.Department of Medicine I, Institute of Cancer ResearchMedical University of ViennaViennaAustria
  10. 10.Institute of Medical GeneticsMedical University of ViennaViennaAustria

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