Metronomic vinorelbine: Anti-angiogenic activity in vitro in normoxic and severe hypoxic conditions, and severe hypoxia-induced resistance to its anti-proliferative effect with reversal by Akt inhibition

Mavroeidis,L.; Sheldon,H.; Briasoulis,E.; Marselos,M.; Pappas,P.; Harris,A. L.

doi:10.3892/ijo.2015.3059

Metronomic vinorelbine: Anti-angiogenic activity in vitro in normoxic and severe hypoxic conditions, and severe hypoxia-induced resistance to its anti-proliferative effect with reversal by Akt inhibition

Authors:
- L. Mavroeidis
- H. Sheldon
- E. Briasoulis
- M. Marselos
- P. Pappas
- A. L. Harris
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Affiliations: Molecular Oncology Laboratories, Weatherall Institute of Molecular Medicine, University of Oxford, John Radcliffe Hospital, Oxford OX3 9DS, UK, Department of Hematology, School of Medicine, University of Ioannina, University Campus, 45110 Ioannina, Greece, Department of Pharmacology, School of Medicine, University of Ioannina, University Campus, 45110 Ioannina, Greece
Published online on: June 19, 2015 https://doi.org/10.3892/ijo.2015.3059
Pages: 455-464
Copyright: © Mavroeidis et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY_NC 3.0].

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Abstract

Metronomic chemotherapy is the protracted, dense administration of low sub-toxic doses of chemotherapy, to inhibit tumor angiogenesis. Vinorelbine is an orally bioavailable vinca alkaloid shown to be useable for metronomic administration. In clinical trials, metronomic vinorelbine has been demonstrated to generate sustainable antitumor efficacy at low nanomolar (nM) concentrations with negligible toxicity. We sought to determine whether the clinically relevant metronomic concentration of vinorelbine is anti-angiogenic in vitro and whether hypoxia, often induced by anti-angiogenic therapy, modifies its effectiveness. We found that the metronomic concentration of 10 nM vinorelbine inhibited human umbilical vein endothelial cell (HUVEC) proliferation, migration, tube formation and sprouting. Severe hypoxia, did not affect the inhibitory effect of metronomic vinorelbine on migration, tube formation and sprouting. However, severe hypoxia reduced its anti-proliferative effect by decreasing its ability to induce G2/M block as it shifted the cell population to the G1 phase and decreased the fraction of the cells in the DNA synthesis S phase. Furthermore, the pro-apoptotic effects of 10 nM vinorelbine were also decreased. Metronomic vinorelbine decreased the Bcl-2/Bax ratio in normoxia whereas the ratio was reduced in severe hypoxia but unaltered by vinorelbine treatment. Akt signals to an anti-apoptotic pathway and we demonstrated that the Akt inhibitor V reversed the protective effect of severe hypoxia. Thus, we provide evidence for the anti-angiogenic basis of metronomic vinorelbine and we show that severe hypoxia mediates resistance to its anti-proliferative effect on endothelial cells. Akt warrants further investigation as a potential target to circumvent this hypoxic resistance.

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1	Kerbel RS and Kamen BA: The anti-angiogenic basis of metronomic chemotherapy. Nat Rev Cancer. 4:423–436. 2004. View Article : Google Scholar : PubMed/NCBI
2	Pasquier E, Kavallaris M and André N: Metronomic chemotherapy: New rationale for new directions. Nat Rev Clin Oncol. 7:455–465. 2010. View Article : Google Scholar : PubMed/NCBI
3	Pasquier E, Honore S and Braguer D: Microtubule-targeting agents in angiogenesis: where do we stand? Drug Resist Update. 9:74–86. 2006. View Article : Google Scholar
4	Briasoulis E, Pappas P, Puozzo C, Tolis C, Fountzilas G, Dafni U, Marselos M and Pavlidis N: Dose-ranging study of metronomic oral vinorelbine in patients with advanced refractory cancer. Clin Cancer Res. 15:6454–6461. 2009. View Article : Google Scholar : PubMed/NCBI
5	Briasoulis E, Aravantinos G, Kouvatseas G, Pappas P, Biziota E, Sainis I, Makatsoris T, Varthalitis I, Xanthakis I, Vassias A, et al: Dose selection trial of metronomic oral vinorelbine monotherapy in patients with metastatic cancer: A hellenic cooperative oncology group clinical translational study. BMC Cancer. 13:2632013. View Article : Google Scholar : PubMed/NCBI
6	Bergers G and Hanahan D: Modes of resistance to anti-angiogenic therapy. Nat Rev Cancer. 8:592–603. 2008. View Article : Google Scholar : PubMed/NCBI
7	Azam F, Mehta S and Harris AL: Mechanisms of resistance to antiangiogenesis therapy. Eur J Cancer. 46:1323–1332. 2010. View Article : Google Scholar : PubMed/NCBI
8	Loges S, Schmidt T and Carmeliet P: Mechanisms of resistance to anti-angiogenic therapy and development of third-generation anti-angiogenic drug candidates. Genes Cancer. 1:12–25. 2010. View Article : Google Scholar : PubMed/NCBI
9	Cosse JP and Michiels C: Tumour hypoxia affects the responsiveness of cancer cells to chemotherapy and promotes cancer progression. Anticancer Agents Med Chem. 8:790–797. 2008. View Article : Google Scholar : PubMed/NCBI
10	Huang L, Ao Q, Zhang Q, Yang X, Xing H, Li F, Chen G, Zhou J, Wang S, Xu G, et al: Hypoxia induced paclitaxel resistance in human ovarian cancers via hypoxia-inducible factor 1alpha. J Cancer Res Clin Oncol. 136:447–456. 2010. View Article : Google Scholar
11	Raz S, Sheban D, Gonen N, Stark M, Berman B and Assaraf YG: Severe hypoxia induces complete antifolate resistance in carcinoma cells due to cell cycle arrest. Cell Death Dis. 5:e10672014. View Article : Google Scholar : PubMed/NCBI
12	Sersa G, Krzic M, Sentjurc M, Ivanusa T, Beravs K, Cemazar M, Auersperg M and Swartz HM: Reduced tumor oxygenation by treatment with vinblastine. Cancer Res. 61:4266–4271. 2001.PubMed/NCBI
13	Zhao D, Jiang L, Hahn EW and Mason RP: Tumor physiologic response to combretastatin A4 phosphate assessed by MRI. Int J Radiat Oncol Biol Phys. 62:872–880. 2005. View Article : Google Scholar : PubMed/NCBI
14	Klement G, Baruchel S, Rak J, Man S, Clark K, Hicklin DJ, Bohlen P and Kerbel RS: Continuous low-dose therapy with vinblastine and VEGF receptor-2 antibody induces sustained tumor regression without overt toxicity. J Clin Invest. 105:R15–R24. 2000. View Article : Google Scholar : PubMed/NCBI
15	Song G, Ouyang G and Bao S: The activation of Akt/PKB signaling pathway and cell survival. J Cell Mol Med. 9:59–71. 2005. View Article : Google Scholar : PubMed/NCBI
16	Rovini A, Savry A, Braguer D and Carré M: Microtubule-targeted agents: When mitochondria become essential to chemotherapy. Biochim Biophys Acta. 1807.679–688. 2011.
17	Kennedy SG, Kandel ES, Cross TK and Hay N: Akt/protein kinase B inhibits cell death by preventing the release of cytochrome c from mitochondria. Mol Cell Biol. 19:5800–5810. 1999.PubMed/NCBI
18	Marty M, Fumoleau P, Adenis A, Rousseau Y, Merrouche Y, Robinet G, Senac I and Puozzo C: Oral vinorelbine pharmacokinetics and absolute bioavailability study in patients with solid tumors. Ann Oncol. 12:1643–1649. 2001. View Article : Google Scholar
19	Potente M, Gerhardt H and Carmeliet P: Basic and therapeutic aspects of angiogenesis. Cell. 146:873–887. 2011. View Article : Google Scholar : PubMed/NCBI
20	Gascoigne KE and Taylor SS: How do anti-mitotic drugs kill cancer cells? J Cell Sci. 122:2579–2585. 2009. View Article : Google Scholar : PubMed/NCBI
21	Gardner LB, Li Q, Park MS, Flanagan WM, Semenza GL and Dang CV: Hypoxia inhibits G1/S transition through regulation of p27 expression. J Biol Chem. 276:7919–7926. 2001. View Article : Google Scholar
22	Chipuk JE and Green DR: How do BCL-2 proteins induce mitochondrial outer membrane permeabilization? Trends Cell Biol. 18:157–164. 2008. View Article : Google Scholar : PubMed/NCBI
23	Bourgarel-Rey V, Savry A, Hua G, Carré M, Bressin C, Chacon C, Imbert J, Braguer D and Barra Y: Transcriptional down-regulation of Bcl-2 by vinorelbine: Identification of a novel binding site of p53 on Bcl-2 promoter. Biochem Pharmacol. 78:1148–1156. 2009. View Article : Google Scholar : PubMed/NCBI
24	Bertolini F, Paul S, Mancuso P, Monestiroli S, Gobbi A, Shaked Y and Kerbel RS: Maximum tolerable dose and low-dose metronomic chemotherapy have opposite effects on the mobilization and viability of circulating endothelial progenitor cells. Cancer Res. 63:4342–4346. 2003.PubMed/NCBI
25	Pasquier E, Tuset MP, Street J, Sinnappan S, MacKenzie KL, Braguer D, Andre N and Kavallaris M: Concentration- and schedule-dependent effects of chemotherapy on the angiogenic potential and drug sensitivity of vascular endothelial cells. Angiogenesis. 16:373–386. 2013. View Article : Google Scholar :
26	Schwartz EL: Antivascular actions of microtubule-binding drugs. Clin Cancer Res. 15:2594–2601. 2009. View Article : Google Scholar : PubMed/NCBI
27	Pourroy B, Honoré S, Pasquier E, Bourgarel-Rey V, Kruczynski A, Briand C and Braguer D: Antiangiogenic concentrations of vinflunine increase the interphase microtubule dynamics and decrease the motility of endothelial cells. Cancer Res. 66:3256–3263. 2006. View Article : Google Scholar : PubMed/NCBI
28	Wang J, Lou P, Lesniewski R and Henkin J: Paclitaxel at ultra low concentrations inhibits angiogenesis without affecting cellular microtubule assembly. Anticancer Drugs. 14:13–19. 2003. View Article : Google Scholar : PubMed/NCBI
29	Calvani M, Rapisarda A, Uranchimeg B, Shoemaker RH and Melillo G: Hypoxic induction of an HIF-1alpha-dependent bFGF autocrine loop drives angiogenesis in human endothelial cells. Blood. 107:2705–2712. 2006. View Article : Google Scholar
30	Ngan VK, Bellman K, Hill BT, Wilson L and Jordan MA: Mechanism of mitotic block and inhibition of cell proliferation by the semisynthetic Vinca alkaloids vinorelbine and its newer derivative vinflunine. Mol Pharmacol. 60:225–232. 2001.PubMed/NCBI
31	Estève MA, Carré M, Bourgarel-Rey V, Kruczynski A, Raspaglio G, Ferlini C and Braguer D: Bcl-2 down-regulation and tubulin subtype composition are involved in resistance of ovarian cancer cells to vinflunine. Mol Cancer Ther. 5:2824–2833. 2006. View Article : Google Scholar : PubMed/NCBI
32	Savry A, Carre M, Berges R, Rovini A, Pobel I, Chacon C, Braguer D and Bourgarel-Rey V: Bcl-2-enhanced efficacy of microtubule-targeting chemotherapy through Bim overexpression: Implications for cancer treatment. Neoplasia. 15:49–60. 2013. View Article : Google Scholar : PubMed/NCBI
33	Zhou H, Li XM, Meinkoth J and Pittman RN: Akt regulates cell survival and apoptosis at a postmitochondrial level. J Cell Biol. 151:483–494. 2000. View Article : Google Scholar : PubMed/NCBI
34	Datta SR, Dudek H, Tao X, Masters S, Fu H, Gotoh Y and Greenberg ME: Akt phosphorylation of BAD couples survival signals to the cell-intrinsic death machinery. Cell. 91:231–241. 1997. View Article : Google Scholar : PubMed/NCBI
35	Majewski N, Nogueira V, Bhaskar P, Coy PE, Skeen JE, Gottlob K, Chandel NS, Thompson CB, Robey RB and Hay N: Hexokinase-mitochondria interaction mediated by Akt is required to inhibit apoptosis in the presence or absence of Bax and Bak. Mol Cell. 16:819–830. 2004. View Article : Google Scholar : PubMed/NCBI
36	Pastorino JG, Hoek JB and Shulga N: Activation of glycogen synthase kinase 3beta disrupts the binding of hexokinase II to mitochondria by phosphorylating voltage-dependent anion channel and potentiates chemotherapy-induced cytotoxicity. Cancer Res. 65:10545–10554. 2005. View Article : Google Scholar : PubMed/NCBI
37	Chiara F, Castellaro D, Marin O, Petronilli V, Brusilow WS, Juhaszova M, Sollott SJ, Forte M, Bernardi P and Rasola A: Hexokinase II detachment from mitochondria triggers apoptosis through the permeability transition pore independent of voltage-dependent anion channels. PloS One. 3:e18522008. View Article : Google Scholar : PubMed/NCBI
38	Brenner C and Grimm S: The permeability transition pore complex in cancer cell death. Oncogene. 25:4744–4756. 2006. View Article : Google Scholar : PubMed/NCBI