Towards the Small and the Beautiful: A Small Dibromotyrosine Derivative from Pseudoceratina sp. Sponge Exhibits Potent Apoptotic Effect through Targeting IKK/NFκB Signaling Pathway
A dibromotyrosine derivative, (1′ R,5′ S,6′ S)-2-(3′,5′-dibromo-1′,6′-dihydroxy-4′-oxocyclohex-2′-enyl) acetonitrile (DT), was isolated from the sponge Pseudoceratina sp., and was found to exhibit a significant cytotoxic activity against leukemia K562 cells. Despite the large number of the isolated bromotyrosine derivatives, studies focusing on their biological mechanism of action are scarce. In the current study we designed a set of experiments to reveal the underlying mechanism of DT cytotoxic activity against K562 cells. First, the results of MTT cytotoxic and the annexin V-FITC/PI apoptotic assays, indicated that the DT cytotoxic activity is mediated through induction of apoptosis. This effect was also supported by caspases-3 and -9 activation as well as PARP cleavage. DT induced generation of reactive oxygen species (ROS) and the disruption of mitochondrial membrane potential (MMP) as indicated by flow cytometric assay. The involvement of ROS generation in the apoptotic activity of DT was further corroborated by the pretreatment of K562 cells with N-acetyl-cysteine (NAC), a ROS scavenger, which prevented apoptosis and the disruption of MMP induced by DT. Results of cell-free system assay suggested that DT can act as a topoisomerase II catalytic inhibitor, unlike the clinical anticancer drug, etoposide, which acts as a topoisomerase poison. Additionally, we found that DT treatment can block IKK/NFκB pathway and activate PI3K/Akt pathway. These findings suggest that the cytotoxic effect of DT is associated with mitochondrial dysfunction-dependent apoptosis which is mediated through oxidative stress. Therefore, DT represents an interesting reference point for the development of new cytotoxic agent targeting IKK/NFκB pathway.
References
[1]
Kunze, K.; Niemann, H.; Ueberlein, S.; Schulze, R.; Ehrlich, H.; Brunner, E.; Proksch, P.; Pee, K.H. Brominated skeletal components of the marine demosponges, Aplysina cavernicola and Ianthella basta: Analytical and biochemical investigations. Mar. Drugs 2013, 11, 1271–1287, doi:10.3390/md11041271.
[2]
Ebada, S.S.; Lin, W.; Proksch, P. Bioactive sesterterpenes and triterpenes from marine sponges: Occurrence and pharmacological significance. Mar. Drugs 2010, 8, 313–346, doi:10.3390/md8020313.
[3]
Lira, N.S.; Montes, R.C.; Tavares, J.F.; da Silva, M.S.; da Cunha, E.V.; de Athayde-Filho, P.F.; Rodrigues, L.C.; da Silva Dias, C.; Barbosa-Filho, J.M. Brominated compounds from marine sponges of the genus Aplysina and a compilation of their 13C NMR spectral data. Mar. Drugs 2011, 9, 2316–2368, doi:10.3390/md9112316.
[4]
Isidori, A. Clinical experimental evaluation of the effect of dibromotyrosine on thyroid function. Int. J. Clin. Pharmacol. Biopharm. 1978, 16, 180–182.
[5]
Galeano, E.; Thomas, O.P.; Robledo, S.; Munoz, D.; Martinez, A. Antiparasitic bromotyrosine derivatives from the marine sponge Verongula rigida. Mar. Drugs 2011, 9, 1902–1913, doi:10.3390/md9101902.
[6]
Hinterding, K.; Knebel, A.; Herrlich, P.; Waldmann, H. Synthesis and biological evaluation of aeroplysinin analogues: A new class of receptor tyrosine kinase inhibitors. Bioorg. Med. Chem. 1998, 6, 1153–1162.
[7]
Cordoba, R.; Tormo, N.S.; Medarde, A.F.; Plumet, J. Antiangiogenic versus cytotoxic activity in analogues of aeroplysinin-1. Bioorg. Med. Chem. 2007, 15, 5300–5315, doi:10.1016/j.bmc.2007.05.011.
[8]
Martinez-Poveda, B.; Garcia-Vilas, J.A.; Cardenas, C.; Melgarejo, E.; Quesada, A.R.; Medina, M.A. The brominated compound aeroplysinin-1 inhibits proliferation and the expression of key pro-inflammatory molecules in human endothelial and monocyte cells. PLoS One 2013, 8, e55203.
[9]
Teeyapant, R.; Woerdenbag, H.J.; Kreis, P.; Hacker, J.; Wray, V.; Witte, L.; Proksch, P. Antibiotic and cytotoxic activity of brominated compounds from the marine sponge Verongia aerophoba. Z. Naturforsch C 1993, 48, 939–945.
[10]
Martinez-Poveda, B.; Rodriguez-Nieto, S.; Garcia-Caballero, M.; Medina, M.A.; Quesada, A.R. The antiangiogenic compound aeroplysinin-1 induces apoptosis in endothelial cells by activating the mitochondrial pathway. Mar. Drugs 2012, 10, 2033–2046, doi:10.3390/md10092033.
[11]
Sallam, A.A.; Ramasahayam, S.; Meyer, S.A.; El Sayed, K.A. Design, synthesis, and biological evaluation of dibromotyrosine analogues inspired by marine natural products as inhibitors of human prostate cancer proliferation, invasion, and migration. Bioorg. Med. Chem. 2010, 18, 7446–7457, doi:10.1016/j.bmc.2010.08.057.
[12]
Kalaitzis, J.A.; Davis, R.A.; Quinn, R.J. Unequivocal 13C NMR assignment of cyclohexadienyl rings in bromotyrosine-derived metabolites from marine sponges. Magn. Reson. Chem. 2012, 50, 749–754, doi:10.1002/mrc.3868.
[13]
Hillgren, J.M.; Oberg, C.T.; Elofsson, M. Syntheses of pseudoceramines A–D and a new synthesis of spermatinamine, bromotyrosine natural products from marine sponges. Org. Biomol. Chem. 2012, 10, 1246–1254, doi:10.1039/c1ob06722b.
[14]
Buchanan, M.S.; Carroll, A.R.; Fechner, G.A.; Boyle, A.; Simpson, M.; Addepalli, R.; Avery, V.M.; Hooper, J.N.; Cheung, T.; Chen, H.; et al. Aplysamine 6, an alkaloidal inhibitor of isoprenylcysteine carboxyl methyltransferase from the sponge Pseudoceratina sp. J. Nat. Prod. 2008, 71, 1066–1067, doi:10.1021/np0706623.
[15]
Kon, Y.; Kubota, T.; Shibazaki, A.; Gonoi, T.; Kobayashi, J. Ceratinadins A–C, new bromotyrosine alkaloids from an Okinawan marine sponge Pseudoceratina sp. Bioorg. Med. Chem. Lett. 2010, 20, 4569–4572, doi:10.1016/j.bmcl.2010.06.015.
[16]
Lebouvier, N.; Jullian, V.; Desvignes, I.; Maurel, S.; Parenty, A.; Dorin-Semblat, D.; Doerig, C.; Sauvain, M.; Laurent, D. Antiplasmodial activities of homogentisic acid derivative protein kinase inhibitors isolated from a Vanuatu marine sponge Pseudoceratina sp. Mar. Drugs 2009, 7, 640–653, doi:10.3390/md7040640.
[17]
Herter-Sprie, G.S.; Chen, S.; Hopker, K.; Reinhardt, H.C. Synthetic lethality as a new concept for the treatment of cancer. Ger. Med. Wkly. 2011, 136, 1526–1530.
[18]
Du, Y.C.; Chang, F.R.; Wu, T.Y.; Hsu, Y.M.; El-Shazly, M.; Chen, C.F.; Sung, P.J.; Lin, Y.Y.; Lin, Y.H.; Wu, Y.C.; et al. Antileukemia component, dehydroeburicoic acid from Antrodia camphorata induces DNA damage and apoptosis in vitro and in vivo models. Phytomedicine 2012, 19, 788–796, doi:10.1016/j.phymed.2012.03.014.
[19]
Nitiss, J.L. Targeting DNA topoisomerase II in cancer chemotherapy. Nat. Rev. Cancer 2009, 9, 338–350, doi:10.1038/nrc2607.
[20]
Sinha, K.; Das, J.; Pal, P.B.; Sil, P.C. Oxidative stress: The mitochondria-dependent and mitochondria-independent pathways of apoptosis. Arch. Toxicol. 2013, 87, 1157–1180, doi:10.1007/s00204-013-1034-4.
[21]
Hutti, J.E.; Pfefferle, A.D.; Russell, S.C.; Sircar, M.; Perou, C.M.; Baldwin, A.S. Oncogenic PI3K mutations lead to NF-kappaB-dependent cytokine expression following growth factor deprivation. Cancer Res. 2012, 72, 3260–3269, doi:10.1158/0008-5472.CAN-11-4141.
[22]
Kim, H.J.; Hawke, N.; Baldwin, A.S. NF-kappaB and IKK as therapeutic targets in cancer. Cell Death Differ. 2006, 13, 738–747, doi:10.1038/sj.cdd.4401877.
[23]
Li, X.; Abdel-Mageed, A.B.; Mondal, D.; Kandil, E. The nuclear factor kappa-B signaling pathway as a therapeutic target against thyroid cancers. Thyroid 2013, 23, 209–218, doi:10.1089/thy.2012.0237.
[24]
Lampiasi, N.; Azzolina, A.; Umezawa, K.; Montalto, G.; McCubrey, J.A.; Cervello, M. The novel NF-kappaB inhibitor DHMEQ synergizes with celecoxib to exert antitumor effects on human liver cancer cells by a ROS-dependent mechanism. Cancer Lett. 2012, 322, 35–44, doi:10.1016/j.canlet.2012.02.008.
[25]
Madonna, G.; Ullman, C.D.; Gentilcore, G.; Palmieri, G.; Ascierto, P.A. NF-kappaB as potential target in the treatment of melanoma. J. Transl. Med. 2012, 10, 53, doi:10.1186/1479-5876-10-53.
[26]
Martelli, A.M.; Tabellini, G.; Bressanin, D.; Ognibene, A.; Goto, K.; Cocco, L.; Evangelisti, C. The emerging multiple roles of nuclear Akt. Biochim. Biophys. Acta 2012, 1823, 2168–2178, doi:10.1016/j.bbamcr.2012.08.017.
[27]
Sawyers, C.L. Chronic myeloid leukemia. N. Engl. J. Med. 1999, 340, 1330–1340, doi:10.1056/NEJM199904293401706.
[28]
Rowley, J.D. Letter: A new consistent chromosomal abnormality in chronic myelogenous leukaemia identified by quinacrine fluorescence and Giemsa staining. Nature 1973, 243, 290–293, doi:10.1038/243290a0.
[29]
Druker, B.J.; Guilhot, F.; O’Brien, S.G.; Gathmann, I.; Kantarjian, H.; Gattermann, N.; Deininger, M.W.; Silver, R.T.; Goldman, J.M.; Stone, R.M.; et al. Five-year follow-up of patients receiving imatinib for chronic myeloid leukemia. N. Engl. J. Med. 2006, 355, 2408–2417, doi:10.1056/NEJMoa062867.
[30]
Ibrahim, A.R.; Eliasson, L.; Apperley, J.F.; Milojkovic, D.; Bua, M.; Szydlo, R.; Mahon, F.X.; Kozlowski, K.; Paliompeis, C.; Foroni, L.; et al. Poor adherence is the main reason for loss of CCyR and imatinib failure for chronic myeloid leukemia patients on long-term therapy. Blood 2011, 117, 3733–3736, doi:10.1182/blood-2010-10-309807.
[31]
Georgiades, S.N.; Clardy, J. Total synthesis of psammaplysenes A and B, naturally occurring inhibitors of FOXO1a nuclear export. Org. Lett. 2005, 7, 4091–4094, doi:10.1021/ol0513286.
[32]
Myatt, S.S.; Brosens, J.J.; Lam, E.W. Sense and sensitivity: FOXO and ROS in cancer development and treatment. Antioxid. Redox Signal. 2011, 14, 675–687, doi:10.1089/ars.2010.3383.
[33]
Greer, E.L.; Brunet, A. FOXO transcription factors at the interface between longevity and tumor suppression. Oncogene 2005, 24, 7410–7425, doi:10.1038/sj.onc.1209086.
[34]
Kryston, T.B.; Georgiev, A.B.; Pissis, P.; Georgakilas, A.G. Role of oxidative stress and DNA damage in human carcinogenesis. Mutat. Res. 2011, 711, 193–201, doi:10.1016/j.mrfmmm.2010.12.016.
[35]
Downward, J. PI 3-kinase, Akt and cell survival. Semin. Cell Dev. Biol. 2004, 15, 177–182, doi:10.1016/j.semcdb.2004.01.002.
[36]
Blanco-Aparicio, C.; Renner, O.; Leal, J.F.; Carnero, A. PTEN, more than the AKT pathway. Carcinogenesis 2007, 28, 1379–1386, doi:10.1093/carcin/bgm052.
[37]
Ahn, J.; Won, M.; Choi, J.H.; Kim, Y.S.; Jung, C.R.; Im, D.S.; Kyun, M.L.; Lee, K.; Song, K.B.; Chung, K.S. Reactive oxygen species-mediated activation of the Akt/ASK1/p38 signaling cascade and p21 downregulation are required for shikonin-induced apoptosis. Apoptosis 2013, 18, 870–881, doi:10.1007/s10495-013-0835-5.
Xing, H.; Weng, D.; Chen, G.; Tao, W.; Zhu, T.; Yang, X.; Meng, L.; Wang, S.; Lu, Y.; Ma, D. Activation of fibronectin/PI-3K/Akt2 leads to chemoresistance to docetaxel by regulating survivin protein expression in ovarian and breast cancer cells. Cancer Lett. 2008, 261, 108–119, doi:10.1016/j.canlet.2007.11.022.
Kishore, N.; Sommers, C.; Mathialagan, S.; Guzova, J.; Yao, M.; Hauser, S.; Huynh, K.; Bonar, S.; Mielke, C.; Albee, L.; et al. A selective IKK-2 inhibitor blocks NF-kappa B-dependent gene expression in interleukin-1 beta-stimulated synovial fibroblasts. J. Biol. Chem. 2003, 278, 32861–32871, doi:10.1074/jbc.M211439200.
[43]
Christopher, J.A.; Avitabile, B.G.; Bamborough, P.; Champigny, A.C.; Cutler, G.J.; Dyos, S.L.; Grace, K.G.; Kerns, J.K.; Kitson, J.D.; Mellor, G.W.; et al. The discovery of 2-amino-3,5-diarylbenzamide inhibitors of IKK-alpha and IKK-beta kinases. Bioorg. Med. Chem. Lett. 2007, 17, 3972–3977, doi:10.1016/j.bmcl.2007.04.088.
[44]
Gamble, C.; McIntosh, K.; Scott, R.; Ho, K.H.; Plevin, R.; Paul, A. Inhibitory kappa B Kinases as targets for pharmacological regulation. Br. J. Pharmacol. 2012, 165, 802–819, doi:10.1111/j.1476-5381.2011.01608.x.
[45]
Nam, S.Y.; Ko, Y.S.; Jung, J.; Yoon, J.; Kim, Y.H.; Choi, Y.J.; Park, J.W.; Chang, M.S.; Kim, W.H.; Lee, B.L. A hypoxia-dependent upregulation of hypoxia-inducible factor-1 by nuclear factor-kappa B promotes gastric tumour growth and angiogenesis. Br. J. Cancer 2011, 104, 166–174, doi:10.1038/sj.bjc.6606020.
[46]
Kuphal, S.; Winklmeier, A.; Warnecke, C.; Bosserhoff, A.K. Constitutive HIF-1 activity in malignant melanoma. Eur. J. Cancer 2010, 46, 1159–1169, doi:10.1016/j.ejca.2010.01.031.
[47]
Crystallographic data have been deposited with the Cambridge Crystallographic Data Centre (deposition number CCDC 930829). Copies of the data can be obtained, f.o.c., on application to the Director, CCDC, 12 Union Road, Cambridge CB2 1EZ, UK (Fax: +44-1223-336033 or E-Mail: deposit@ccdc.cam.ac.uk).
[48]
Capon, R.; Macleod, J. Two epimeric dibromo nitriles from the australian sponge Aplysina laevis. Aust. J. Chem. 1987, 40, 341–346, doi:10.1071/CH9870341.
[49]
Santalova, E.A.; Denisenko, V.A.; Glazunov, V.P.; Kalinovskii, A.I.; Anastyuk, S.D.; Stonik, V.A. Dibromotyrosine derivatives from the ethanol extract of the marine sponge Aplysina sp.: Structures, transformations and origin. Russian Chem. Bull. 2100, 60, 570–580.
[50]
Huang, K.-J.; Chen, Y.-C.; El-Shazly, M.; Du, Y.-C.; Su, J.-H.; Tsao, C.-W.; Yen, W.-H.; Chang, W.-B.; Su, Y.-D.; Yeh, Y.-T.; et al. 5-Episinuleptolide acetate, a norcembranoidal diterpene from the formosan soft coral Sinularia sp., induces leukemia cell apoptosis through Hsp90 inhibition. Molecules 2013, 18, 2924–2933, doi:10.3390/molecules18032924.
