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01.12.2015 | Original Paper

Role of JNK and NF-κB in mediating the effect of combretastatin A-4 and brimamin on endothelial and carcinoma cells

verfasst von: Katharina Mahal, Aamir Ahmad, Seema Sethi, Marcus Resch, Ralf Ficner, Fazlul H. Sarkar, Rainer Schobert, Bernhard Biersack

Erschienen in: Cellular Oncology | Ausgabe 6/2015

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Abstract

Purpose

The 4,5-diarylimidazole brimamin is an analog of the natural vascular-disrupting agent combretastatin A-4 (CA-4) with improved water solubility, tolerance by animals and efficacy in multidrug-resistant tumors. Here, we aimed at identifying the major mechanisms underlying the in vitro and in vivo actions of brimamin on endothelial and carcinoma cells, including vascularization.

Methods

The contribution of specific signaling kinases to the effects of brimamin on cytoskeleton organization and the viability and differentiation of endothelial cells was assessed by MTT and tube formation assays in the presence or absence of specific kinase inhibitors. Changes in DNA affinity and expression of NF-κB in endothelial and carcinoma-derived cells and their solid tumors (xenografts) treated with brimamin were ascertained by electrophoretic mobility shift assays and Western blotting. The anti-vascular effect of brimamin in solid tumors was verified by CD31 immunostaining.

Results

We found that brimamin can inhibit tubulin polymerization and cause a reorganization of F-actin in Ea.hy926 endothelial cells. Its inhibitory effect on tube formation was found to depend on functional Rho kinase and JNK. JNK inhibition was found to suppress the induction of endothelial cell apoptosis by brimamin. In CA-4-refractory human BxPC-3 pancreas carcinoma-derived and triple-negative MDA-MB-231 breast carcinoma-derived cells brimamin was found to inhibit growth and to induce apoptosis at low nanomolar concentrations by blocking NF-κB activation in a dose-dependent manner. Brimamin was also found to reduce the in vivo growth rate and vascularization of MDA-MB-231 xenografts in mice. Residual tumor cells of these treated xenografts showed a relatively low expression of the p65 subunit of NF-κB.

Conclusions

Our data indicate that cellular JNK and Rho kinase activities are crucial for the cytotoxic and cytoskeleton reorganizing effects of brimamin on endothelial cells. In addition, we found that in resistant carcinoma cells and xenografts brimamin can induce down-regulation of anti-apoptotic NF-κB expression and signaling. Its chemical properties and efficacy against clinically relevant cancer entities make brimamin a promising candidate vascular-disrupting agent.

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G. Pettit, S. Singh, E. Hamel, C. Lin, D. Alberts, D. Garcia-Kendall, Isolation and structure of the strong cell growth and tubulin inhibitor combretastatin A-4. Experientia 45, 209–211 (1989)CrossRefPubMed

C.J. Mooney, G. Nagaiah, P. Fu, J.K. Wasman, M.M. Cooney, P.S. Savvides, J.A. Bokar, A. Dowlati, D. Wang, S.S. Agarwala, A phase II trial of fosbretabulin in advanced anaplastic thyroid carcinoma and correlation of baseline serum-soluble intracellular adhesion molecule-1 with outcome. Thyroid 19, 233–240 (2009)PubMedCentralCrossRefPubMed

G.J. Rustin, G. Shreeves, P.D. Nathan, A. Gaya, T.S. Ganesan, D. Wang, J. Boxall, L. Poupard, D.J. Chaplin, M.R.L. Stratford, J. Balkissoon, M. Zweifel, A Phase Ib trial of CA4P (combretastatin A-4 phosphate), carboplatin, and paclitaxel in patients with advanced cancer. Br. J. Cancer 102, 1355–1360 (2010)PubMedCentralCrossRefPubMed

G.C. Tron, T. Pirali, G. Sorba, F. Pagliai, S. Busacca, A.A. Genazzani, Medicinal chemistry of combretastatin A4: present and future directions. J. Med. Chem. 49, 3033–3044 (2006)CrossRefPubMed

S.E. Holwell, P.A. Cooper, M.J. Thompson, G.R. Pettit, L.W. Lippert 3rd, S.W. Martin, M.C. Bibby, Anti-tumor and anti-vascular effects of the novel tubulin-binding agent combretastatin A-1 phosphate. Anticancer Res. 22, 3933–3940 (2002)PubMed

L.K. Folkes, M. Christlieb, E. Madej, M.R.L. Stratford, P. Wardman, Oxidative metabolism of combretastatin A-1 produces quinone intermediates with the potential to bind to nucleophiles and to enhance oxidative stress via free radicals. Chem. Res. Toxicol. 20, 1885–1894 (2007)CrossRefPubMed

G.R. Pettit, M.R. Rhodes, D.L. Herald, D.J. Chaplin, M.R. Stratford, E. Hamel, R.K. Pettit, J.C. Chapuis, D. Oliva, Antineoplastic agents 393. Synthesis of the trans-isomer of combretastatin A-4 prodrug. Anticancer Drug Des. 13, 981–993 (1998)PubMed

G.R. Pettit, B.E. Toki, D.L. Herald, M.R. Boyd, E. Hamel, R.K. Pettit, J.C. Chapuis, Antineoplastic agents. 410. Asymmetric hydroxylation of trans-combretastatin A-4. J. Med. Chem. 42, 1459–1465 (1999)CrossRefPubMed

L. Wang, K.W. Woods, Q. Li, K.J. Barr, R.W. McCroskey, S.M. Hannick, L. Gherke, R.B. Credo, Y.-H. Hui, K. Marsh, R. Warner, J.Y. Lee, N. Zielinski-Mozng, D. Frost, S.H. Rosenberg, H.L. Sham, Potent, orally active heterocycle-based combretastatin A-4 analogues: synthesis, structure − activity relationship, pharmacokinetics, and in vivo antitumor activity evaluation. J. Med. Chem. 45, 1697–1711 (2002)CrossRefPubMed

10.

R. Schobert, B. Biersack, A. Dietrich, K. Effenberger, S. Knauer, T. Mueller, 4-(3-Halo/amino-4,5-dimethoxyphenyl)-5-aryloxazoles and -N-methylimidazoles that are cytotoxic against combretastatin A resistant tumor cells and vascular disrupting in a cisplatin resistant germ cell tumor model. J. Med. Chem. 53, 6595–6602 (2010)CrossRefPubMed

11.

K. Mahal, B. Biersack, H. Caysa, R. Schobert, T. Mueller, Combretastatin A-4 derived imidazoles show cytotoxic, antivascular, and antimetastatic effects based on cytoskeletal reorganisation. Investig. New Drugs 33, 541–554 (2015)CrossRef

12.

C. Kanthou, The tumor vascular targeting agent combretastatin A-4-phosphate induces reorganization of the actin cytoskeleton and early membrane blebbing in human endothelial cells. Blood 99, 2060–2069 (2002)CrossRefPubMed

13.

B.B. Aggarwal, S. Shishodia, S.K. Sandur, M.K. Pandey, G. Sethi, Inflammation and cancer: how hot is the link? Biochem. Pharmacol. 72, 1605–1621 (2006)CrossRefPubMed

14.

A. Ahmad, S. Banerjee, Z. Wang, D. Kong, F.H. Sarkar, Plumbagin-induced apoptosis of human breast cancer cells is mediated by inactivation of NF-κB and Bcl-2. J. Cell. Biochem. 105, 1461–1471 (2008)CrossRefPubMed

15.

S. Ali, A. Ahmad, S. Banerjee, S. Padhye, K. Dominiak, J.M. Schaffert, Z. Wang, P.A. Philip, F.H. Sarkar, Gemcitabine sensitivity can be induced in pancreatic cancer cells through modulation of miR-200 and miR-21 expression by curcumin or its analogue CDF. Cancer Res. 70, 3606–3617 (2010)PubMedCentralCrossRefPubMed

16.

Y. He, H. Xu, L. Liang, Z. Zhan, X. Yang, X. Yu, Y. Ye, L. Sun, Antiinflammatory effect of Rho kinase blockade via inhibition of NF-kappaB activation in rheumatoid arthritis. Arthritis Rheum. 58, 3366–3376 (2008)CrossRefPubMed

17.

H. Shimada, L.E. Rajagopalan, Rho kinase-2 activation in human endothelial cells drives lysophosphatidic acid-mediated expression of cell adhesion molecules via NF-kappaB p65. J. Biol. Chem. 285, 12536–12542 (2010)PubMedCentralCrossRefPubMed

18.

S. Shimizu, M. Tahara, S. Ogata, K. Hashimoto, K. Morishige, K. Tasaka, Y. Murata, Involvement of nuclear factor-kappaB activation through RhoA/Rho-kinase pathway in LPS-induced IL-8 production in human cervical stromal cells. Mol. Hum. Reprod. 13, 181–187 (2007)CrossRefPubMed

19.

G.G. Mackenzie, C.L. Keen, P.I. Oteiza, Microtubules are required for NF-kappaB nuclear translocation in neuroblastoma IMR-32 cells: modulation by zinc. J. Neurochem. 99, 402–415 (2006)CrossRefPubMed

20.

V. Bourgarel-Rey, S. Vallee, O. Rimet, S. Champion, D. Braguer, A. Desobry, C. Briand, Y. Barra, Involvement of nuclear factor kappaB in c-Myc induction by tubulin polymerization inhibitors. Mol. Pharmacol. 59, 1165–1170 (2001)PubMed

21.

T.F. Hansen, B.S. Nielsen, A. Jakobsen, F.B. Sorensen, Visualising and quantifying angiogenesis in metastatic colorectal cancer. A comparison of methods and their predictive value for chemotherapy response. Cell. Oncol. 36, 341–350 (2013)CrossRef

22.

B. Pula, M. Olbromski, A. Wojnar, A. Gomulkiewicz, W. Witkiewicz, M. Ugorski, P. Dziegiel, M. Podhorska-Okolow, Impact of SOX18 expression in cancer cells and vessels on the outcome of invasive ductal breast carcinoma. Cell. Oncol. 36, 469–483 (2013)CrossRef

23.

A.W. Schüttelkopf, D.M.F. van Aalten, PRODRG: a tool for high-throughput crystallography of protein-ligand complexes. Acta Crystallogr. D Biol. Crystallogr. 60, 1355–1363 (2004)CrossRefPubMed

24.

O. Trott, A.J. Olson, AutoDock Vina: improving the speed and accuracy of docking with a new scoring function, efficient optimization, and multithreading. J. Comput. Chem. 31, 455–461 (2009)

25.

J. Gasteiger, M. Marsili, Iterative partial equalization of orbital electronegativity – a rapid access to atomic charges. Tetrahedron 36, 3219–3228 (1980)CrossRef

26.

G.L. Warren, C.W. Andrews, A.-M. Capelli, B. Clarke, J. LaLonde, M.H. Lambert, M. Lindvall, N. Nevins, S.F. Semus, S. Senger, G. Tedesco, I.D. Wall, J.M. Woolven, C.E. Peishoff, M.S. Head, A critical assessment of docking programs and scoring functions. J. Med. Chem. 49, 5912–5931 (2006)CrossRefPubMed

27.

W. DeLano, The PyMOL molecular graphics system (DeLano Sci, LLC San Carlos CA, 2003)

28.

K. Mahal, S. Schruefer, G. Steinemann, F. Rausch, R. Schobert, M. Höpfner, B. Biersack, Biological evaluation of 4,5-diarylimidazoles with hydroxamic acid appendages as novel dual mode anticancer agents. Cancer Chemother. Pharmacol. 75, 691–700 (2015)CrossRefPubMed

29.

K. Mahal, M. Resch, R. Ficner, R. Schobert, B. Biersack, T. Mueller, Effects of the tumor-vasculature-disrupting agent verubulin and two heteroaryl analogues on cancer cells, endothelial cells, and blood vessels. ChemMedChem 9, 847–854 (2014)CrossRefPubMed

30.

G.M. Tozer, C. Kanthou, B.C. Baguley, Disrupting tumour blood vessels. Nat. Rev. Cancer 5, 423–435 (2005)CrossRefPubMed

31.

B. Biersack, K. Effenberger, R. Schobert, M. Ocker, Oxazole-bridged combretastatin a analogues with improved anticancer properties. ChemMedChem 5, 420–427 (2010)CrossRefPubMed

32.

S. Ali, Simultaneous targeting of the epidermal growth factor receptor and cyclooxygenase-2 pathways for pancreatic cancer therapy. Mol. Cancer Ther. 4, 1943–1951 (2005)CrossRefPubMed

33.

R.B.G. Ravelli, B. Gigant, P.A. Curmi, I. Jourdain, S. Lachkar, A. Sobel, M. Knossow, Insight into tubulin regulation from a complex with colchicine and a stathmin-like domain. Nature 428, 198–202 (2004)CrossRefPubMed

34.

M. Botta, S. Forli, M. Magnani, F. Manetti, Molecular modeling approaches to study the binding mode on tubulin of microtubule destabilizing and stabilizing agents. Top. Curr. Chem. 286, 279–328 (2009)CrossRefPubMed

35.

C. Kanthou, G.M. Tozer, Tumour targeting by microtubule-depolymerising vascular disrupting agents. Expert Opin. Ther. Targets 11, 1443–1457 (2007)CrossRefPubMed

36.

H. Quan, Y. Xu, L. Lou, p38 MAPK, but not ERK1/2, is critically involved in the cytotoxicity of the novel vascular disrupting agent combretastatin A4. Int. J. Cancer 122, 1730–1737 (2007)CrossRef

37.

M. Fan, L. Du, A. Stone, K. Gilbert, T. Chambers, Modulation of mitogen-activated protein kinases and phosphorylation of Bcl-2 by vinblastine represent persistent forms of normal fluctuations at G2-M. Cancer Res. 60, 6403–6407 (2000)PubMed

38.

C. Kanthou, G.M. Tozer, Microtubule depolymerizing vascular disrupting agents: novel therapeutic agents for oncology and other pathologies. Int. J. Exp. Pathol. 90, 284–294 (2009)PubMedCentralCrossRefPubMed

39.

M. Chrzanowska-Wodnicka, K. Burridge, Rho-stimulated contractility drives the formation of stress fibers and focal adhesions. J. Cell Biol. 133, 1403–1415 (1996)CrossRefPubMed

40.

L.J. Williams, D. Mukherjee, M. Fisher, C.C. Reyes-Aldasoro, S. Akerman, C. Kanthou, G.M. Tozer, An in vivo role for Rho kinase activation in the tumour vascular disrupting activity of combretastatin A-4 3-O-phosphate: Rho kinase and tumour vascular targeting. Br. J. Pharmacol. 171, 4902–4913 (2014)PubMedCentralCrossRefPubMed

41.

A. Arlt, A. Gehrz, S. Müerköster, J. Vorndamm, M.-L. Kruse, U.R. Fölsch, H. Schäfer, Role of NF-kappaB and Akt/PI3K in the resistance of pancreatic carcinoma cell lines against gemcitabine-induced cell death. Oncogene 22, 3243–3251 (2003)CrossRefPubMed

42.

M. Graupera, J. Guillermet-Guibert, L.C. Foukas, L.-K. Phng, R.J. Cain, A. Salpekar, W. Pearce, S. Meek, J. Millan, P.R. Cutillas, A.J.H. Smith, A.J. Ridley, C. Ruhrberg, H. Gerhardt, B. Vanhaesebroeck, Angiogenesis selectively requires the p110α isoform of PI3K to control endothelial cell migration. Nature 453, 662–666 (2008)CrossRefPubMed

43.

J. Karar, A. Maity, PI3K/AKT/mTOR pathway in angiogenesis. Front. Mol. Neurosci. 4, 51 (2011)PubMedCentralCrossRefPubMed

44.

G. Rajashekhar, M. Kamocka, A. Marin, M.A. Suckow, W.R. Wolter, S. Badve, A.R. Sanjeevaiah, K. Pumiglia, E. Rosen, M. Clauss, Pro-inflammatory angiogenesis is mediated by p38 MAP kinase. J. Cell. Physiol. 226, 800–808 (2011)CrossRefPubMed

45.

M. Kelkel, C. Cerella, F. Mack, T. Schneider, C. Jacob, M. Schumacher, M. Dicato, M. Diederich, ROS-independent JNK activation and multisite phosphorylation of Bcl-2 link diallyl tetrasulfide-induced mitotic arrest to apoptosis. Carcinogenesis 33, 2162–2171 (2012)CrossRefPubMed

46.

J.D. Orth, A. Loewer, G. Lahav, T.J. Mitchison, Prolonged mitotic arrest triggers partial activation of apoptosis, resulting in DNA damage and p53 induction. Mol. Biol. Cell 23, 567–576 (2012)PubMedCentralCrossRefPubMed

47.

S.P. Tabruyn, A.W. Griffioen, A new role for NF-kappaB in angiogenesis inhibition. Cell Death Differ. 14, 1393–1397 (2007)CrossRefPubMed

48.

S.P. Tabruyn, S. Memet, P. Ave, C. Verhaeghe, K.H. Mayo, I. Struman, J.A. Martial, A.W. Griffioen, NF-kappaB activation in endothelial cells is critical for the activity of angiostatic agents. Mol. Cancer Ther. 8, 2645–2654 (2009)CrossRefPubMed

49.

M. Carr, L.M. Greene, A.J.S. Knox, D.G. Lloyd, D.M. Zisterer, M.J. Meegan, Lead identification of conformationally restricted β-lactam type combretastatin analogues: synthesis, antiproliferative activity and tubulin targeting effects. Eur. J. Med. Chem. 45, 5752–5766 (2010)CrossRefPubMed

50.

H. Wehbe, C.M. Kearney, K.G. Pinney, Combretastatin A-4 resistance in H460 human lung carcinoma demonstrates distinctive alterations in β-tubulin isotype expression. Anticancer Res. 25, 3865–3870 (2005)PubMed

51.

E.C. Tampaki, L. Nakopoulou, A. Tampakis, K. Kontzoglou, W.P. Weber, G. Kouraklis, Nestin involvement in tissue injury and cancer - a potential tumor marker? Cell. Oncol. 37, 305–315 (2014)CrossRef

52.

D.O. Moon, M.O. Kim, C.H. Kang, J.D. Lee, Y.H. Choi, G.Y. Kim, JNK inhibitor SP600125 promotes the formation of polymerized tubulin, leading to G2/M phase arrest, endoreduplication, and delayed apoptosis. Exp. Mol. Med. 41, 665–677 (2009)PubMedCentralCrossRefPubMed

53.

A.V. Singh, M. Bandi, N. Raje, P. Richardson, M.A. Palladino, D. Chauhan, K.C. Anderson, A novel vascular disrupting agent plinabulin triggers JNK-mediated apoptosis and inhibits angiogenesis in multiple myeloma cells. Blood 117, 5692–5700 (2011)PubMedCentralCrossRefPubMed

54.

L. Ciani, P.C. Salinas, c-Jun N-terminal kinase (JNK) cooperates with Gsk3β to regulate dishevelled-mediated microtubule stability. BMC Cell Biol. 8, 27 (2007)PubMedCentralCrossRefPubMed

55.

D.C.H. Ng, T.T. Zhao, Y.Y.C. Yeap, K.R. Ngoei, M.A. Bogoyevitch, c-Jun N-terminal kinase phosphorylation of stathmin confers protection against cellular stress. J. Biol. Chem. 285, 29001–29013 (2010)PubMedCentralCrossRefPubMed

56.

Y.Y. Yip, Y.Y.C. Yeap, M.A. Bogoyevitch, D.C.H. Ng, Differences in c-Jun N-terminal kinase recognition and phosphorylation of closely related stathmin-family members. Biochem. Biophys. Res. Commun. 446, 248–254 (2014)CrossRefPubMed

57.

J.-H. Kim, T.H. Kim, H.S. Kang, J. Ro, H.S. Kim, S. Yoon, SP600125, an inhibitor of Jnk pathway, reduces viability of relatively resistant cancer cells to doxorubicin. Biochem. Biophys. Res. Commun. 387, 450–455 (2009)CrossRefPubMed

58.

X. Wang, K. Belguise, N. Kersual, K.H. Kirsch, N.D. Mineva, F. Galtier, D. Chalbos, G.E. Sonenshein, Oestrogen signalling inhibits invasive phenotype by repressing RelB and its target BCL2. Nat. Cell Biol. 9, 470–478 (2007)PubMedCentralCrossRefPubMed

59.

H. Lee, J. Jeon, Y.S. Ryu, J.E. Jeong, S. Shin, T. Zhang, S.W. Kang, J.H. Hong, G.M. Hur, Disruption of microtubules sensitizes the DNA damage-induced apoptosis through inhibiting nuclear factor kappaB (NF-κB) DNA-binding activity. J. Korean Med. Sci. 25, 1574 (2010)PubMedCentralCrossRefPubMed

60.

S. Papa, Linking JNK signaling to NF-kappaB: a key to survival. J. Cell Sci. 117, 5197–5208 (2004)CrossRefPubMed

61.

L. Li, B.B. Aggarwal, S. Shishodia, J. Abbruzzese, R. Kurzrock, Nuclear factor-kappaB and IkappaB kinase are constitutively active in human pancreatic cells, and their down-regulation by curcumin (diferuloylmethane) is associated with the suppression of proliferation and the induction of apoptosis. Cancer 101, 2351–2362 (2004)CrossRefPubMed

Titel: Role of JNK and NF-κB in mediating the effect of combretastatin A-4 and brimamin on endothelial and carcinoma cells
verfasst von: Katharina Mahal
Aamir Ahmad
Seema Sethi
Marcus Resch
Ralf Ficner
Fazlul H. Sarkar
Rainer Schobert
Bernhard Biersack
Publikationsdatum: 01.12.2015
Verlag: Springer Netherlands
Erschienen in: Cellular Oncology / Ausgabe 6/2015
Print ISSN: 2211-3428
Elektronische ISSN: 2211-3436
DOI: https://doi.org/10.1007/s13402-015-0243-7

Springer Medizin

Role of JNK and NF-κB in mediating the effect of combretastatin A-4 and brimamin on endothelial and carcinoma cells

Abstract

Purpose

Methods

Results

Conclusions

Neu im Fachgebiet Pathologie

Molekularpathologische Untersuchungen im Wandel der Zeit

Vergleichende Pathologie in der onkologischen Forschung

Gastrointestinale Stromatumoren

Personalisierte Medizin in der Onkologie

Springer Medizin

Abstract

Purpose

Methods

Results

Conclusions

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