Multiple Active Compounds from Viscum album L. Synergistically Converge to Promote Apoptosis in Ewing Sarcoma
Monika Twardziok, Susann Kleinsimon, Jana Rolff, Sebastian Jäger, Angelika Eggert, Georg Seifert, Catharina I Delebinski, Monika Twardziok, Susann Kleinsimon, Jana Rolff, Sebastian Jäger, Angelika Eggert, Georg Seifert, Catharina I Delebinski
Abstract
Ewing sarcoma is the second most common bone cancer in children and adolescents, with poor prognosis and outcome in ~70% of initial diagnoses and 10-15% of relapses. Hydrophobic triterpene acids and hydrophilic lectins and viscotoxins from European mistletoe (Viscum album L.) demonstrate anticancer properties, but have not yet been investigated for Ewing sarcoma. Commercial Viscum album L. extracts are aqueous, excluding the insoluble triterpenes. We recreated a total mistletoe effect by combining an aqueous extract (viscum) and a triterpene extract (TT) solubilized with cyclodextrins. Ewing sarcoma cells were treated with viscum, TT and viscumTT in vitro, ex vivo and in vivo. In vitro and ex vivo treatment of Ewing sarcoma cells with viscum inhibited proliferation and induced apoptosis in a dose-dependent fashion, while viscumTT combination treatment generated a synergistic effect. Apoptosis occurred via intrinsic and extrinsic apoptotic pathways, evidenced by activation of both CASP8 and CASP9. We show that viscumTT treatment shifts the balance of apoptotic regulatory proteins towards apoptosis, mainly via CLSPN, MCL1, BIRC5 and XIAP downregulation. ViscumTT also demonstrated strong antitumor activity in a cell line- and patient-derived mouse model, and may be considered an adjuvant therapy option for pediatric patients with Ewing sarcoma.
Conflict of interest statement
Competing Interests: The authors have the following interests: Jana Rolff is employed by EPO GmbH and Sebastian Jäger by Birken AG. There are no patents, products in development or marketed products to declare. This does not alter the authors' adherence to all the PLOS ONE policies on sharing data and materials, as detailed online in the guide for authors.
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References
- Society AC (2014) American Cancer Society: Cancer Facts and Figures 2014. American Cancer Society, available online.
- Kaatsch P, Spix C (2014) German Childhood Cancer Registry—Report 2013/14 (1980–2013). Institute of Medical Biostatistics, Epidemiology and Informatics (IMBEI) at the University Medical Center of the Johannes Gutenberg University Mainz.
- von Levetzow C, Jiang X, Gwye Y, von Levetzow G, Hung L, et al. (2011) Modeling Initiation of Ewing Sarcoma in Human Neural Crest Cells. PLoS ONE 6: e19305 10.1371/journal.pone.0019305
- Lin PP, Wang Y, Lozano G (2011) Mesenchymal Stem Cells and the Origin of Ewing's Sarcoma. Sarcoma 2011.
- Paronetto MP (2013) Ewing Sarcoma Protein: A Key Player in Human Cancer. International Journal of Cell Biology 2013: 12.
- Mackintosh C, Madoz-Gurpide J (2013) Mining Sarcomas by Proteomics Approaches: Ewing Sarcoma on the Spotlight. Recent Pat Biotechnol 7: 98–111.
- Amaral AT, Ordóñez JL, Otero-Motta AP, García-Domínguez DJ, Sevillano MV, et al. (2014) Innovative Therapies in Ewing Sarcoma. Advances in Anatomic Pathology 21: 44–62. 10.1097/PAP.0000000000000003
- Stahl M, Ranft A, Paulussen M, Bölling T, Vieth V, et al. (2011) Risk of recurrence and survival after relapse in patients with Ewing sarcoma. Pediatric Blood & Cancer 57: 549–553.
- Hingorani P, Dickman P, Garcia-Filion P, White-Collins A, Kolb EA, et al. (2013) BIRC5 expression is a poor prognostic marker in Ewing sarcoma. Pediatr Blood Cancer 60: 35–40. 10.1002/pbc.24290
- Greve B, Sheikh-Mounessi F, Kemper B, Ernst I, Gotte M, et al. (2012) Survivin, a target to modulate the radiosensitivity of Ewing's sarcoma. Strahlenther Onkol 188: 1038–1047. 10.1007/s00066-012-0223-z
- Coumar MS, Tsai F-Y, Kanwar JR, Sarvagalla S, Cheung CHA (2013) Treat cancers by targeting survivin: Just a dream or future reality? Cancer Treatment Reviews 39: 802–811. 10.1016/j.ctrv.2013.02.002
- Kashkar H (2010) X-linked Inhibitor of Apoptosis: A Chemoresistance Factor or a Hollow Promise. Clinical Cancer Research 16: 4496–4502. 10.1158/1078-0432.CCR-10-1664
- Obexer P, Ausserlechner MJ (2014) X-Linked Inhibitor of Apoptosis Protein–A Critical Death Resistance Regulator and Therapeutic Target for Personalized Cancer Therapy. Frontiers in Oncology 4: 197 10.3389/fonc.2014.00197
- Marvibaigi M, Supriyanto E, Amini N, Abdul Majid FA, Jaganathan SK (2014) Preclinical and Clinical Effects of Mistletoe against Breast Cancer. Biomed Res Int 2014: 785479 10.1155/2014/785479
- Franz H, Ziska P, Kindt A (1981) Isolation and properties of three lectins from mistletoe (Viscum album L.). BiochemJ 195: 481–484.
- Orhan DD, Kupeli E, Yesilada E, Ergun F (2006) Anti-inflammatory and antinociceptive activity of flavonoids isolated from Viscum album ssp. album. ZNaturforschC 61: 26–30.
- Amer B, Juvik OJ, Francis GW, Fossen T (2013) Novel GHB-derived natural products from European mistletoe (Viscum album). Pharm Biol 51: 981–986. 10.3109/13880209.2013.773520
- Nhiem NX, Kiem PV, Minh CV, Kim N, Park S, et al. (2013) Diarylheptanoids and flavonoids from viscum album inhibit LPS-stimulated production of pro-inflammatory cytokines in bone marrow-derived dendritic cells. J Nat Prod 76: 495–502. 10.1021/np300490v
- Jung ML, Baudino S, Ribereau-Gayon G, Beck JP (1990) Characterization of cytotoxic proteins from mistletoe (Viscum album L.). Cancer Lett 51: 103–108.
- Maletzki C, Linnebacher M, Savai R, Hobohm U (2013) Mistletoe lectin has a shiga toxin-like structure and should be combined with other Toll-like receptor ligands in cancer therapy. Cancer Immunol Immunother 62: 1283–1292. 10.1007/s00262-013-1455-1
- Klingbeil MF, Xavier FC, Sardinha LR, Severino P, Mathor MB, et al. (2013) Cytotoxic effects of mistletoe (Viscum album L.) in head and neck squamous cell carcinoma cell lines. Oncol Rep 30: 2316–2322. 10.3892/or.2013.2732
- Ucar EO, Arda N, Aitken A (2012) Extract from mistletoe, Viscum album L., reduces Hsp27 and 14-3-3 protein expression and induces apoptosis in C6 rat glioma cells. Genet Mol Res 11: 2801–2813.
- von Schoen-Angerer T, Goyert A, Vagedes J, Kiene H, Merckens H, et al. (2014) Disappearance of an advanced adenomatous colon polyp after intratumoural injection with Viscum album (European mistletoe) extract: a case report. J Gastrointestin Liver Dis 23: 449–452. 10.15403/jgld.2014.1121.234.acpy
- Jäger S, Winkler K, Pfüller U, Scheffler A (2007) Solubility studies of oleanolic acid and betulinic acid in aqueous solutions and plant extracts of Viscum album L. Planta Med 73: 157–162.
- Akl MR, Elsayed HE, Ebrahim HY, Haggag EG, Kamal AM, et al. (2014) 3-O-[N-(p-fluorobenzenesulfonyl)-carbamoyl]-oleanolic acid, a semisynthetic analog of oleanolic acid, induces apoptosis in breast cancer cells. Eur J Pharmacol 740: 209–217. 10.1016/j.ejphar.2014.07.011
- Petronelli A, Saulle E, Pasquini L, Petrucci E, Mariani G, et al. (2009) High sensitivity of ovarian cancer cells to the synthetic triterpenoid CDDO-Imidazolide. Cancer Lett 282: 214–228. 10.1016/j.canlet.2009.03.018
- Lucio KA, Rocha Gda G, Moncao-Ribeiro LC, Fernandes J, Takiya CM, et al. (2011) Oleanolic acid initiates apoptosis in non-small cell lung cancer cell lines and reduces metastasis of a B16F10 melanoma model in vivo. PLoS One 6: e28596 10.1371/journal.pone.0028596
- Fulda S, Jeremias I, Pietsch T, Debatin KM (1999) Betulinic acid: a new chemotherapeutic agent in the treatment of neuroectodermal tumors. Klin Padiatr 211: 319–322.
- Soica C, Oprean C, Borcan F, Danciu C, Trandafirescu C, et al. (2014) The synergistic biologic activity of oleanolic and ursolic acids in complex with hydroxypropyl-gamma-cyclodextrin. Molecules 19: 4924–4940. 10.3390/molecules19044924
- Delebinski CI, Jäger S, Kemnitz-Hassanin K, Henze G, Lode HN, et al. (2012) A new development of triterpene acid-containing extracts from Viscum album L. displays synergistic induction of apoptosis in acute lymphoblastic leukaemia. Cell Proliferation 45: 176–187. 10.1111/j.1365-2184.2011.00801.x
- Delebinski CI, Twardziok M, Kleinsimon S, Hoff F, Mulsow K, et al. (2015) A Natural Combination Extract of Viscum album L. Containing Both Triterpene Acids and Lectins Is Highly Effective against AML In Vivo. PLoS ONE 10: e0133892 10.1371/journal.pone.0133892
- Strüh CM, Jäger S, Kersten A, Schempp CM, Scheffler A, et al. (2013) Triterpenoids amplify anti-tumoral effects of mistletoe extracts on murine B16.f10 melanoma in vivo. PLoS One 8: e62168 10.1371/journal.pone.0062168
- Strüh CM, Jäger S, Schempp CM, Scheffler A, Martin SF (2012) A Novel Triterpene Extract from Mistletoe Induces Rapid Apoptosis in Murine B16.F10 Melanoma Cells. Phytotherapy Research 26: 1507–1512. 10.1002/ptr.4604
- Jaggy C, Musielski H, Urech K, Schaller G (1995) Quantitative determination of lectins in mistletoe preparations. Arzneimittelforschung 45: 905–909.
- Green DR, Kroemer G (2004) The pathophysiology of mitochondrial cell death. Science 305: 626–629.
- Decaudin D, Marzo I, Brenner C, Kroemer G (1998) Mitochondria in chemotherapy-induced apoptosis: a prospective novel target of cancer therapy (review). Int J Oncol 12: 141–152.
- Ehrhardt H, Hofig I, Wachter F, Obexer P, Fulda S, et al. (2012) NOXA as critical mediator for drug combinations in polychemotherapy. Cell DeathDis 3: e327.
- Foungbé S, Kouassi G, Kablan JB, Marcy R (1991) Study of Costus lucanusianus: plant juice, fraction combinations and pharmacologie estimation of natural product total activity. Journal of Ethnopharmacology 33: 221–226.
- Ji H-F, Li X-J, Zhang H-Y (2009) Natural products and drug discovery. Can thousands of years of ancient medical knowledge lead us to new and powerful drug combinations in the fight against cancer and dementia? EMBO Reports 10: 194–200. 10.1038/embor.2009.12
- Bantel H, Engels IH, Voelter W, Schulze-Osthoff K, Wesselborg S (1999) Mistletoe lectin activates caspase-8/FLICE independently of death receptor signaling and enhances anticancer drug-induced apoptosis. Cancer Res 59: 2083–2090.
- Kim MS, So HS, Lee KM, Park JS, Lee JH, et al. (2000) Activation of caspase cascades in Korean mistletoe (Viscum album var. coloratum) lectin-II-induced apoptosis of human myeloleukemic U937 cells. GenPharmacol 34: 349–355.
- Yang X, Jiang S, Liu Y, Zhang P, Xie S, et al. (2012) Recombinant VAA-I from Viscum album induces apoptotic cell death of hepatocellular carcinoma SMMC7721 cells. Molecules 17: 11435–11446. 10.3390/molecules171011435
- Timoshenko AV, Gorudko IV, Andre S, Gabius HJ (2000) Cell-type dependence of stability modulation of lectin-initiated contacts by impairment of multivalent carbohydrate binding and intracellular signaling. Biosci Rep 20: 199–209.
- Ito Y, Pandey P, Sporn MB, Datta R, Kharbanda S, et al. (2001) The novel triterpenoid CDDO induces apoptosis and differentiation of human osteosarcoma cells by a caspase-8 dependent mechanism. MolPharmacol 59: 1094–1099.
- Chakravarti B, Maurya R, Siddiqui JA, Bid HK, Rajendran SM, et al. (2012) In vitro anti-breast cancer activity of ethanolic extract of Wrightia tomentosa: role of pro-apoptotic effects of oleanolic acid and urosolic acid. J Ethnopharmacol 142: 72–79.
- Yan SL, Huang CY, Wu ST, Yin MC (2010) Oleanolic acid and ursolic acid induce apoptosis in four human liver cancer cell lines. ToxicolIn Vitro 24: 842–848.
- Shyu M-H, Kao T-C, Yen G-C (2010) Oleanolic Acid and Ursolic Acid Induce Apoptosis in HuH7 Human Hepatocellular Carcinoma Cells through a Mitochondrial-Dependent Pathway and Downregulation of XIAP. Journal of Agricultural and Food Chemistry 58: 6110–6118. 10.1021/jf100574j
- Wei J, Liu M, Liu H, Wang H, Wang F, et al. (2013) Oleanolic acid arrests cell cycle and induces apoptosis via ROS-mediated mitochondrial depolarization and lysosomal membrane permeabilization in human pancreatic cancer cells. J Appl Toxicol 33: 756–765. 10.1002/jat.2725
- Konopleva M, Tsao T, Estrov Z, Lee R-m, Wang R-Y, et al. (2004) The Synthetic Triterpenoid 2-Cyano-3,12-dioxooleana-1,9-dien-28-oic Acid Induces Caspase-Dependent and -Independent Apoptosis in Acute Myelogenous Leukemia. Cancer Research 64: 7927–7935.
- He X, Wang Y, Hu H, Zhang Z (2012) In vitro and in vivo antimammary tumor activities and mechanisms of the apple total triterpenoids. J Agric Food Chem 60: 9430–9436.
- Park YK, Do YR, Jang BC (2012) Apoptosis of K562 leukemia cells by Abnobaviscum F(R), a European mistletoe extract. Oncol Rep 28: 2227–2232. 10.3892/or.2012.2026
- Ryu K, Susa M, Choy E, Yang C, Hornicek FJ, et al. (2010) Oleanane triterpenoid CDDO-Me induces apoptosis in multidrug resistant osteosarcoma cells through inhibition of Stat3 pathway. BMCCancer 10: 187.
- Leal AS, Wang R, Salvador JA, Jing Y (2013) Synthesis of novel heterocyclic oleanolic acid derivatives with improved antiproliferative activity in solid tumor cells. Org Biomol Chem 11: 1726–1738. 10.1039/c3ob00011g
- May WA, Grigoryan RS, Keshelava N, Cabral DJ, Christensen LL, et al. (2013) Characterization and Drug Resistance Patterns of Ewing's Sarcoma Family Tumor Cell Lines. PLoS ONE 8: e80060 10.1371/journal.pone.0080060
- Büssing A, Multani AS, Pathak S, Pfüller U, Schietzel M (1998) Induction of apoptosis by the N-acetyl-galactosamine-specific toxic lectin from Viscum album L. is associated with a decrease of nuclear p53 and Bcl-2 proteins and induction of telomeric associations. Cancer Lett 130: 57–68.
- Choi SH, Yang H, Lee SH, Ki J-H, Nam D-H, et al. (2014) TopBP1 and Claspin contribute to the radioresistance of lung cancer brain metastases. Molecular Cancer 13: 211 10.1186/1476-4598-13-211
- Lopez J, Meier P (2010) To fight or die: inhibitor of apoptosis proteins at the crossroad of innate immunity and death. Current Opinion in Cell Biology 22: 872–881. 10.1016/j.ceb.2010.08.025
- de Almagro MC, Vucic D (2012) The inhibitor of apoptosis (IAP) proteins are critical regulators of signaling pathways and targets for anti-cancer therapy. Exp Oncol 34: 200–211.
- Duan Z, Ames R, Ryan M, Hornicek FJ, Mankin H, et al. (2009) CDDO-Me, a synthetic triterpenoid, inhibits expression of IL-6 and Stat3 phosphorylation in multi-drug resistant ovarian cancer cells. Cancer chemotherapy and pharmacology 63: 681–689. 10.1007/s00280-008-0785-8
- Bache M, Bernhardt S, Passin S, Wichmann H, Hein A, et al. (2014) Betulinic Acid Derivatives NVX-207 and B10 for Treatment of Glioblastoma—An in Vitro Study of Cytotoxicity and Radiosensitization. International Journal of Molecular Sciences 15: 19777–19790. 10.3390/ijms151119777
- Huber R, Eisenbraun J, Miletzki B, Adler M, Scheer R, et al. (2010) Pharmacokinetics of natural mistletoe lectins after subcutaneous injection. Eur J Clin Pharmacol 66: 889–897. 10.1007/s00228-010-0830-5
- Frantz M, Jung ML, Ribereau-Gayon G, Anton R (2000) Modulation of mistletoe (Viscum album L.) lectins cytotoxicity by carbohydrates and serum glycoproteins. Arzneimittelforschung 50: 471–478.
- Duong Van Huyen JP, Delignat S, Bayry J, Kazatchkine MD, Bruneval P, et al. (2006) Interleukin-12 is associated with the in vivo anti-tumor effect of mistletoe extracts in B16 mouse melanoma. Cancer Lett 243: 32–37.
- Seifert G, Jesse P, Laengler A, Reindl T, Luth M, et al. (2008) Molecular mechanisms of mistletoe plant extract-induced apoptosis in acute lymphoblastic leukemia in vivo and in vitro. Cancer Lett 264: 218–228. 10.1016/j.canlet.2008.01.036
- Braun JM, Ko HL, Schierholz JM, Beuth J (2002) Standardized mistletoe extract augments immune response and down-regulates local and metastatic tumor growth in murine models. Anticancer Res 22: 4187–4190.
- Hua Y, Zhang Z, Li J, Li Q, Hu S, et al. (2009) Oleanolic acid derivative Dex-OA has potent anti-tumor and anti-metastatic activity on osteosarcoma cells in vitro and in vivo. Investigational New Drugs 29: 258–265. 10.1007/s10637-009-9354-1
- Liby K, Risingsong R, Royce DB, Williams CR, Ma T, et al. (2009) Triterpenoids CDDO-methyl ester or CDDO-ethyl amide and rexinoids LG100268 or NRX194204 for prevention and treatment of lung cancer in mice. Cancer PrevRes(Phila) 2: 1050–1058.
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