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The anti-invasive activity of synthetic alkaloid ethoxyfagaronine on L1210 leukemia cells is mediated by down-regulation of plasminogen activators and MT1-MMP expression and activity

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Summary

Quaternary benzo[c]phenanthridines such as fagaronine are natural substances which have been reported to exhibit anticancer and anti-leukemic properties. However, the therapeutic use of these molecules is limited due to the high dose required to exhibit anti-tumor activity and subsequent toxicity. In this study, we describe the therapeutic potential of a new derivative of fagaronine, Ethoxyfagaronine (N-methyl-12-ethoxy-2hydroxy-3, 8, 9-trimethoxybenzo[c]-phenanthridiniumchlorhydrate) as an anti-leukemic agent. Cytotoxic activity and cell growth inhibition of Ethoxyfagaronine (Etxfag) was tested on murine L1210 leukemia cells using trypan blue assay and MTT assay. At the concentration of 10−7 M, Etxfag induced less than 10% of cell death. Etxfag (10−7 M) was tested on L1210 cell invasiveness using matrigel™ precoated transwell chambers and efficiently reduces the invasive potential of L1210 cells by more than 50% as compared with untreated cells. Western blot and immunofluorescence experiments showed that Etxfag decreased both MT1-MMP expression and activation at the cell surface, decreased plasmin activity by down-regulating u-PAR and uPA expression at the cell surface and increasing PAI-1 secretion in conditioned media. The set of our findings underscore the therapeutic potential of ethoxyfagaronine as a new potential anticancer agent able to prevent leukemic cell dissemination.

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Acknowledgements

I would like to thank Dr H. Kaplan for his help with Analyze and Amira. I would like also to thank Dr L. Devy for her advices.

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Authors and Affiliations

  1. UMR CNRS 6237, Laboratoire SiRMa, IFR 53 Interactions Cellules Microenvironnement, UFR Sciences Exactes et Naturelles, Moulin de la Housse, Université de Reims Champagne Ardenne (URCA), BP 1039, F-51687, Reims Cedex 2, France

    Jérôme Devy, Farid Ouchani, Christelle Oudot, Stéphanie Salesse, Fanja Rabenoelina, Siana Al-Khara, Isabelle Letinois, Laurent Martiny & Emmanuelle Charpentier

  2. Laboratoire de Chimie Organique IFR QUASAV 149 UFR Sciences UFR Sciences Pharmaceutiques et Ingénierie de la Santé, Université d’Angers, SONAS, 16 bd Daviers, 49045, ANGERS Cedex, France

    Jean Jacques Helesbeux, Enguerran Vanquelef & Olivier Duval

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  1. Jérôme Devy
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Correspondence to Jérôme Devy.

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This work was supported by grants from the Centre National de la Recherche Scientifique (UMR 6237), the University of Reims Champagne Ardenne, the Region Champagne Ardenne, the Ligue Nationale Contre le Cancer (Comité de la Haute-Marne). The confocal microscope observations were performed at INRA, UMR FARE, Reims.

Jérôme Devy and Farid Ouchani have equally contributed to the work.

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Devy, J., Ouchani, F., Oudot, C. et al. The anti-invasive activity of synthetic alkaloid ethoxyfagaronine on L1210 leukemia cells is mediated by down-regulation of plasminogen activators and MT1-MMP expression and activity. Invest New Drugs 29, 730–741 (2011). https://doi.org/10.1007/s10637-010-9410-x

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