Summary
Angiogenesis plays a pivotal role in tumorigenesis and also contributes to the pathogenesis of hematologic malignancies. A number of plant compounds have shown efficacy in preclinical and clinical studies and some of them possess an anti-angiogenic activity. Our present findings report anti-angiogenic activities of ethoxyfagaronine (etxfag), a synthetic derivative of fagaronine. Once determined the non-cytotoxic concentration of etxfag, we showed that the drug inhibits VEGF-induced angiogenesis in a Matrigel™ plug assay and suppresses ex vivo sprouting from VEGF-treated aortic rings. Each feature leading to neovascularization was then investigated and results demonstrate that etxfag prevents VEGF-induced migration and tube formation in human umbilical vein endothelial cells (HUVEC). Moreover, etxfag also suppresses VEGF-induced VEGFR-2 phosphorylation and inhibits FAK phosphorylation at Y-861 as well as focal adhesion complex turnover. Beside these effects, etxfag modifies MT1-MMP localization at the endothelial cell membrane. Finally, immunoprecipitation assay revealed that etxfag decreases VEGF binding to VEGFR-2. As we previously reported that etxfag is able to prevent leukemic cell invasiveness and adhesion to fibronectin, all together our data collectively support the anti-angiogenic activities of etxfag which could represent an additional approach to current anti-cancer therapies.
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Shibuya M (2006) Vascular endothelial growth factor receptor-1 (VEGFR-1/Flt-1): a dual regulator for angiogenesis. Angiogenesis 9:225–230, discussion 31
Shibuya M (2014) VEGF-VEGFR signals in health and disease. Biomol Ther (Seoul) 22:1–9
Cross MJ, Dixelius J, Matsumoto T, Claesson-Welsh L (2003) VEGF-receptor signal transduction. Trends Biochem Sci 28:488–494
Kampen KR, Ter Elst A, de Bont ES (2013) Vascular endothelial growth factor signaling in acute myeloid leukemia. Cell Mol Life Sci 70:1307–1317
Podar K, Anderson KC (2005) The pathophysiologic role of VEGF in hematologic malignancies: therapeutic implications. Blood 105:1383–1395
Podar K, Anderson KC (2010) A therapeutic role for targeting c-Myc/Hif-1-dependent signaling pathways. Cell Cycle 9:1722–1728
Medinger M, Mross K (2010) Clinical trials with anti-angiogenic agents in hematological malignancies. J Angiogenes Res 2:10
Lucas DM, Still PC, Perez LB, Grever MR, Kinghorn AD (2010) Potential of plant-derived natural products in the treatment of leukemia and lymphoma. Curr Drug Targets 11:812–822
Basini G, Bussolati S, Santini SE, Grasselli F (2009) Stanniocalcin, a potential ovarian angiogenesis regulator, does not affect endothelial cell apoptosis. Ann N Y Acad Sci 1171:94–99
Eun JP, Koh GY (2004) Suppression of angiogenesis by the plant alkaloid, sanguinarine. Biochem Biophys Res Commun 317:618–624
De Stefano I, Raspaglio G, Zannoni GF, Travaglia D, Prisco MG, Mosca M et al (2009) Antiproliferative and antiangiogenic effects of the benzophenanthridine alkaloid sanguinarine in melanoma. Biochem Pharmacol 78:1374–1381
Messmer WM, Tin-Wa M, Fong HH, Bevelle C, Farnsworth NR, Abraham DJ et al (1972) Fagaronine, a new tumor inhibitor isolated from Fagara zanthoxyloides Lam. (Rutaceae). J Pharm Sci 61:1858–1859
Comoe L, Jeannesson P, Trentesaux C, Desoize B, Jardillier JC (1987) The antileukemic alkaloid fagaronine and the human K 562 leukemic cells: effects on growth and induction of erythroid differentiation. Leuk Res 11:445–451
Stermitz FR, Larson KA, Kim DK (1973) Some structural relationships among cytotoxic and antitumor benzophenanthridine alkaloid derivatives. J Med Chem 16:939–940
Lynch MA, Duval O, Sukhanova A, Devy J, MacKay SP, Waigh RD et al (2001) Synthesis, biological activity and comparative analysis of DNA binding affinities and human DNA topoisomerase I inhibitory activities of novel 12-alkoxy-benzo[c]phenanthridinium salts. Bioorg Med Chem Lett 11:2643–2646
Deroussent A, Re M, Hoellinger H, Vanquelef E, Duval O, Sonnier M et al (2004) In vitro metabolism of ethoxidine by human CYP1A1 and rat microsomes: identification of metabolites by high-performance liquid chromatography combined with electrospray tandem mass spectrometry and accurate mass measurements by time-of-flight mass spectrometry. Rapid Commun Mass Spectrom 18:474–482
Workman P, Aboagye EO, Balkwill F, Balmain A, Bruder G, Chaplin DJ et al (2010) Guidelines for the welfare and use of animals in cancer research. Br J Cancer 102:1555–1577
Ouchani F, Devy J, Rusciani A, Helesbeux JJ, Salesse S, Letinois I et al (2012) Targeting focal adhesion assembly by ethoxyfagaronine prevents lymphoblastic cell adhesion to fibronectin. Anal Cell Pathol 35:267–284
Abu-Ghazaleh R, Kabir J, Jia H, Lobo M, Zachary I (2001) Src mediates stimulation by vascular endothelial growth factor of the phosphorylation of focal adhesion kinase at tyrosine 861, and migration and anti-apoptosis in endothelial cells. Biochem J 360:255–264
Galvez BG, Matias-Roman S, Albar JP, Sanchez-Madrid F, Arroyo AG (2001) Membrane type 1-matrix metalloproteinase is activated during migration of human endothelial cells and modulates endothelial motility and matrix remodeling. J Biol Chem 276:37491–37500
Medinger M, Fischer N, Tzankov A (2010) Vascular endothelial growth factor-related pathways in hemato-lymphoid malignancies. J Oncol 2010:729725
Sallam TH, El Telbany MA, Mahmoud HM, Iskander MA (2013) Significance of neuropilin-1 expression in acute myeloid leukemia. Turk J Haematol 30:300–306
Ma J, Waxman DJ (2008) Combination of antiangiogenesis with chemotherapy for more effective cancer treatment. Mol Cancer Ther 7:3670–3684
Thevenard J, Ramont L, Devy J, Brassart B, Dupont-Deshorgue A, Floquet N et al (2010) The YSNSG cyclopeptide derived from tumstatin inhibits tumor angiogenesis by down-regulating endothelial cell migration. Int J Cancer J Int Du Cancer 126:1055–1066
Genis L, Galvez BG, Gonzalo P, Arroyo AG (2006) MT1-MMP: universal or particular player in angiogenesis? Cancer Metastasis Rev 25:77–86
Lauffenburger DA, Horwitz AF (1996) Cell migration: a physically integrated molecular process. Cell 84:359–369
Parat MO, Anand-Apte B, Fox PL (2003) Differential caveolin-1 polarization in endothelial cells during migration in two and three dimensions. Mol Biol Cell 14:3156–3168
Tahir SA, Park S, Thompson TC (2009) Caveolin-1 regulates VEGF-stimulated angiogenic activities in prostate cancer and endothelial cells. Cancer Biol Ther 8:2286–2296
Neufeld G, Cohen T, Gengrinovitch S, Poltorak Z (1999) Vascular endothelial growth factor (VEGF) and its receptors. FASEB J 13:9–22
Devy J, Ouchani F, Oudot C, Helesbeux JJ, Vanquelef E, Salesse S, et al (2010). 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
Acknowledgments
Plateforme IBiSA PICT “Imagerie Cellulaire et Tissulaire” This work was supported by grants from CNRS and Ligue Contre le Cancer Grand Est. We would like to thank J.L. Breda for animal care. The authors acknowledge Dr Laetitia Devy Dimanche for editorial assistance.
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Ouchani, F., Jeanne, A., Thevenard, J. et al. Ethoxyfagaronine, a synthetic analogue of fagaronine that inhibits vascular endothelial growth factor-1, as a new anti-angiogeneic agent. Invest New Drugs 33, 75–85 (2015). https://doi.org/10.1007/s10637-014-0184-4
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DOI: https://doi.org/10.1007/s10637-014-0184-4