Cancer Chemotherapy and Pharmacology

, Volume 54, Issue 5, pp 432–440 | Cite as

Inhibition of HUVEC tubulogenesis by hederacolchiside-A1 is associated with plasma membrane cholesterol sequestration and activation of the Ha-Ras/MEK/ERK cascade

  • Chantal Barthomeuf
  • Dominique Boivin
  • Richard Béliveau
Original Article
First Online:
Received:
Accepted:

Abstract

Purpose

Neoangiogenesis is critical to cancer proliferation and metastasis and constitutes an attractive target for cancer therapy. It has previously been demonstrated that hederacolchiside-A1 (HCol-A1), a triterpenoid saponin from Hedera colchica Koch, has antimelanoma potential. The goal of this study was to evaluate, in vitro, if in addition to its tumoricidal effect on melanoma cells, HCol-A1 might affect endothelial cell network formation.

Methods

We investigated whether HCol-A1 affects matrigel-induced tubulogenesis and inhibits the viability (WST-1 assay) of human umbilical vein endothelial cells (HUVECs). To provide structure-activity relationships (SAR), studies were conducted on HCol-A1, oleanolic acid and hederacolchiside A (HCol-A), a triterpenoid saponin which possess the same sugar sequence as Hcol-A1. Plasma membrane cholesterol sequestration was studied by labelling with [3H]cholesterol and assayed with HCol-A1-cholesterol complexes. HCol-A1 signalling was investigated using immunoassays.

Results

In contrast to HCol-A and oleanolic acid, HCol-A1 inhibited matrigel-induced angiogenesis at micromolar concentration. Plasma membrane cholesterol sequestration was found to be critical for this activity. Activation of the Ras/MEK/ERK cascade appears to be one of the mechanisms by which Hcol-A1 affects HUVEC network formation. The predominant activation of the Ha-Ras isoform, which decreases HUVEC-tolerance to apoptosis, might contribute to the high susceptibility of this cell line to HCol-A1.

Conclusion

Since cholesterol sequestration affects cell confluence-dependent remodelling of endothelial membranes and vascular endothelial growth factor receptor-2 activity, these results raise the possibility that Hcol-A1 might slow-down cancer proliferation and metastasis in vivo by inhibiting critical aspects of neoangiogenesis. Further in vivo studies are needed to verify this hypothesis.

Keywords

Cancer Angiogenesis Triterpene saponin Hederacolchiside-A1 Cholesterol ERK Ras 

Abbreviations

CC

Column chromatography

DMEM

Dulbecco’s modified Eagle’s medium

EGCg

Epigallocatechin gallate

FBS

Fetal bovine serum

HCol-A1

Hederacolchiside A1

HCol-A

Hederacolchiside A

Ha-Ras

Harvey isoform of Ras

Ki-Ras

Kirsten isoform of Ras

mAb

Monoclonal antibody

pAb

Polyclonal antibody

PBS

Phosphate-buffered saline

PVDF

Polyvinylidene difluoride

RP-HPLC

Reverse-phase high-pressure liquid chromatography

ROS

Reactive oxygen species

SAK

Extracellular signal-regulated kinase

VEGFR-2

Vascular endothelial growth factor receptor-2

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Notes

Acknowledgements

The authors thank Nicole Lafontaine for excellent technical assistance. They are grateful to Prof. G. Balansard for the generous gift of oleanolic acid and to Dr. V. Mishvildadze for prepurified hederacolchiside-A1 and hederacolchiside-A samples. They also thank Dr. Michel Demeule at Université du Québec à Montréal (UQAM) for his critical reading of the manuscript. This work was in part supported by a grant from the Canadian Institute of Health Research to Richard Béliveau.

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Copyright information

© Springer-Verlag 2004

Authors and Affiliations

  • Chantal Barthomeuf
    • 1
  • Dominique Boivin
    • 2
  • Richard Béliveau
    • 2
  1. 1.Laboratoire de Pharmacognosie et Biotechnologies, Faculté de PharmacieUMR-INSERM U-484-UdA-CJPClermont-Fd CedexFrance
  2. 2.Laboratoire de Médecine Moléculaire, Hôpital Ste-Justine-UQAM, Centre de Cancérologie Charles BruneauCentre de Recherche de l’Hôpital Ste-JustineMontréalCanada

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