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Planta Med 2013; 79(12): 1017-1023
DOI: 10.1055/s-0032-1328715
Biological and Pharmacological Activity
Original Papers
Georg Thieme Verlag KG Stuttgart · New York

A Hydroalcoholic Extract from the Leaves of Nerium oleander Inhibits Glycolysis and Induces Selective Killing of Lung Cancer Cells

José Manuel Calderón-Montaño
1   Department of Pharmacology, Faculty of Pharmacy, University of Seville, Seville, Spain
,
Estefanía Burgos-Morón
1   Department of Pharmacology, Faculty of Pharmacy, University of Seville, Seville, Spain
,
Manuel Luis Orta
2   Department of Cell Biology, Faculty of Biology, University of Seville, Seville, Spain
,
Santiago Mateos
2   Department of Cell Biology, Faculty of Biology, University of Seville, Seville, Spain
,
Miguel López-Lázaro
1   Department of Pharmacology, Faculty of Pharmacy, University of Seville, Seville, Spain
› Author Affiliations
Further Information

Publication History

received 25 April 2013
revised 24 May 2013

accepted 31 May 2013

Publication Date:
03 July 2013 (online)

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Abstract

Recent evidence suggests that cardiac glycosides might be used for the treatment of cancer. The ornamental shrub Nerium oleander has been used in traditional medicine for treating several disorders including cancer, and extracts from the leaves of this plant have already entered phase I clinical trials. In this communication, we have prepared a hydroalcoholic extract from the leaves of Nerium oleander (containing 4.75 ± 0.32 % of cardenolides) and have assessed its cytotoxic activity in A549 lung cancer cells vs. MRC5 nonmalignant lung fibroblasts. The results showed that the cytotoxicity of the Nerium oleander extract against the cancer cell line was significantly higher than that against the nonmalignant cell line, with a potency and selectivity similar to those of the anticancer drug cisplatin. Pretreatment of A549 cells with the antioxidants N-acetylcysteine and catalase slightly prevented the cytotoxicity of the extract, therefore suggesting that the formation of reactive oxygen species participates in its cytotoxic activity but does not play a major role. Nerium oleander extract-induced cytotoxicity and DNA damage (gamma-H2AX focus formation) were slightly higher in cells lacking BRCA2 (deficient in homologous recombination repair) than in parental cells; this indicates that the induction of DNA damage may also play a role in the cytotoxicity of the extract. Nerium oleander extract induced a marked inhibition of glycolysis (glucose consumption and lactate production) in A549 cells, comparable to that of the glycolysis inhibitor dichloroacetate (currently in clinical development for cancer therapy). Because platinum compounds are widely used in the treatment of lung cancer, we tested the cytotoxicity of several combinations of cisplatin with the extract and found a moderate synergism when Nerium oleander extract was administered after cisplatin but a moderate antagonism when it was added before cisplatin. Our results suggest that extracts from Nerium oleander might induce anticancer effects in patients with lung cancer and support their possible advancement into phase II clinical trials for the treatment of this type of cancer.

Key words

Nerium oleander - Apocynaceae - cardiac glycosides - cardiotonic steroids - anticancer
 

  • References

  • 1 Rahimtoola SH, Tak T. The use of digitalis in heart failure. Curr Probl Cardiol 1996; 21: 781-853
    CrossrefPubMedGoogle Scholar
  • 2 Xie Z, Askari A. Na(+)/K(+)-ATPase as a signal transducer. Eur J Biochem 2002; 269: 2434-2439
    CrossrefPubMedGoogle Scholar
  • 3 Schoner W, Scheiner-Bobis G. Endogenous and exogenous cardiac glycosides: their roles in hypertension, salt metabolism, and cell growth. Am J Physiol Cell Physiol 2007; 293: C509-C536
    CrossrefPubMedGoogle Scholar
  • 4 Haux J. Digitoxin is a potential anticancer agent for several types of cancer. Med Hypotheses 1999; 53: 543-548
    CrossrefPubMedGoogle Scholar
  • 5 Lopez-Lazaro M. Digitoxin as an anticancer agent with selectivity for cancer cells: possible mechanisms involved. Expert Opin Ther Targets 2007; 11: 1043-1053
    CrossrefPubMedGoogle Scholar
  • 6 Newman RA, Yang P, Pawlus AD, Block KI. Cardiac glycosides as novel cancer therapeutic agents. Mol Interv 2008; 8: 36-49
    CrossrefPubMedGoogle Scholar
  • 7 Prassas I, Diamandis EP. Novel therapeutic applications of cardiac glycosides. Nat Rev Drug Discov 2008; 7: 926-935
    CrossrefPubMedGoogle Scholar
  • 8 Mijatovic T, Van Quaquebeke E, Delest B, Debeir O, Darro F, Kiss R. Cardiotonic steroids on the road to anti-cancer therapy. Biochim Biophys Acta 2007; 1776: 32-57
    PubMedGoogle Scholar
  • 9 Sreenivasan Y, Raghavendra PB, Manna SK. Oleandrin-mediated expression of Fas potentiates apoptosis in tumor cells. J Clin Immunol 2006; 26: 308-322
    CrossrefPubMedGoogle Scholar
  • 10 Hallbook H, Felth J, Eriksson A, Fryknas M, Bohlin L, Larsson R, Gullbo J. Ex vivo activity of cardiac glycosides in acute leukaemia. PLoS One 2011; 6: e15718
    CrossrefPubMedGoogle Scholar
  • 11 Lopez-Lazaro M, Pastor N, Azrak SS, Ayuso MJ, Austin CA, Cortes F. Digitoxin inhibits the growth of cancer cell lines at concentrations commonly found in cardiac patients. J Nat Prod 2005; 68: 1642-1645
    CrossrefPubMedGoogle Scholar
  • 12 Mijatovic T, Op DB, Van Quaquebeke E, Dewelle J, Darro F, de Launoit Y, Kiss R. The cardenolide UNBS1450 is able to deactivate nuclear factor kappaB-mediated cytoprotective effects in human non-small cell lung cancer cells. Mol Cancer Ther 2006; 5: 391-399
    CrossrefPubMedGoogle Scholar
  • 13 Zhang H, Qian DZ, Tan YS, Lee K, Gao P, Ren YR, Rey S, Hammers H, Chang D, Pili R, Dang CV, Liu JO, Semenza GL. Digoxin and other cardiac glycosides inhibit HIF-1alpha synthesis and block tumor growth. Proc Natl Acad Sci USA 2008; 105: 19579-19586
    CrossrefPubMedGoogle Scholar
  • 14 Lopez-Lazaro M. Digoxin, HIF-1, and cancer. Proc Natl Acad Sci USA 2009; 106: E26
    CrossrefPubMedGoogle Scholar
  • 15 Rashan LJ, Franke K, Khine MM, Kelter G, Fiebig HH, Neumann J, Wessjohann LA. Characterization of the anticancer properties of monoglycosidic cardenolides isolated from Nerium oleander and Streptocaulon tomentosum . J Ethnopharmacol 2011; 134: 781-788
    CrossrefPubMedGoogle Scholar
  • 16 Mijatovic T, Kiss R. Cardiotonic steroids-mediated Na+/K+-ATPase targeting could circumvent various chemoresistance pathways. Planta Med 2013; 79: 189-198
    Article in Thieme ConnectPubMedGoogle Scholar
  • 17 Perne A, Muellner MK, Steinrueck M, Craig-Mueller N, Mayerhofer J, Schwarzinger I, Sloane M, Uras IZ, Hoermann G, Nijman SM, Mayerhofer M. Cardiac glycosides induce cell death in human cells by inhibiting general protein synthesis. PLoS One 2009; 4: e8292
    CrossrefPubMedGoogle Scholar
  • 18 Pelicano H, Martin DS, Xu RH, Huang P. Glycolysis inhibition for anticancer treatment. Oncogene 2006; 25: 4633-4646
    CrossrefPubMedGoogle Scholar
  • 19 Lopez-Lazaro M. The Warburg effect: why and how do cancer cells activate glycolysis in the presence of oxygen?. Anticancer Agents Med Chem 2008; 8: 305-312
    CrossrefPubMedGoogle Scholar
  • 20 Lopez-Lazaro M, Palma DLP, Pastor N, Martin-Cordero C, Navarro E, Cortes F, Ayuso MJ, Toro MV. Anti-tumour activity of Digitalis purpurea L. subsp. heywoodii . Planta Med 2003; 69: 701-704
    Article in Thieme ConnectPubMedGoogle Scholar
  • 21 Watters GP, Smart DJ, Harvey JS, Austin CA. H2AX phosphorylation as a genotoxicity endpoint. Mutat Res 2009; 679: 50-58
    PubMedGoogle Scholar
  • 22 Lobrich M, Shibata A, Beucher A, Fisher A, Ensminger M, Goodarzi AA, Barton O, Jeggo PA. gammaH2AX foci analysis for monitoring DNA double-strand break repair: strengths, limitations and optimization. Cell Cycle 2010; 9: 662-669
    CrossrefPubMedGoogle Scholar
  • 23 Pastor N, Pinero J, Ortiz T, Mateos JC, De Miguel M, Cortes F. Topoisomerase activities and levels in irradiated Chinese hamster AA8 cells and in its radiosensitive mutant EM9. Int J Radiat Biol 1999; 75: 1035-1042
    CrossrefPubMedGoogle Scholar
  • 24 Cao X, Bloomston M, Zhang T, Frankel WL, Jia G, Wang B, Hall NC, Koch RM, Cheng H, Knopp MV, Sun D. Synergistic antipancreatic tumor effect by simultaneously targeting hypoxic cancer cells with HSP90 inhibitor and glycolysis inhibitor. Clin Cancer Res 2008; 14: 1831-1839
    CrossrefPubMedGoogle Scholar
  • 25 Mekhail T, Kaur H, Ganapathi R, Budd GT, Elson P, Bukowski RM. Phase 1 trial of Anvirzel in patients with refractory solid tumors. Invest New Drugs 2006; 24: 423-427
    CrossrefPubMedGoogle Scholar
  • 26 Henary HA, Kurzrock R, Falchook GS, Naing A, Moulder SL, Wheler JJ, Tsimberidou AM, Durand J, Yang P, Johansen MJ, Newman R, Khan R, Patel U, Hong DS. Final results of a first-in-human phase I trial of PBI-05204, an inhibitor of AKT, FGF-2, NF-Kb, and p 70S6K in advanced cancer patients. ASCO Meeting Abstracts 2011; 29: 3023
    PubMedGoogle Scholar
  • 27 Lopez-Lazaro M, Calderon-Montano JM, Burgos-Moron E, Austin CA. Green tea constituents (−)-epigallocatechin-3-gallate (EGCG) and gallic acid induce topoisomerase I- and topoisomerase II-DNA complexes in cells mediated by pyrogallol-induced hydrogen peroxide. Mutagenesis 2011; 26: 489-498
    CrossrefPubMedGoogle Scholar
  • 28 Vega-Perez JM, Palo-Nieto C, Periñan I, Vega-Holm M, Calderon-Montano JM, Lopez-Lazaro M, Iglesias-Guerra F. Stereoselective dihydroxylation reaction of alkenyl β-D-hexopyranosides: a methodology for the synthesis of glycosylglycerol derivatives and 1-O-acyl-3-O-β-D-glycosyl-sn-glycerol analogues. Eur J Org Chem 2012; 6: 1237-1252
    PubMedGoogle Scholar
  • 29 Alexandre J, Batteux F, Nicco C, Chereau C, Laurent A, Guillevin L, Weill B, Goldwasser F. Accumulation of hydrogen peroxide is an early and crucial step for paclitaxel-induced cancer cell death both in vitro and in vivo . Int J Cancer 2006; 119: 41-48
    CrossrefPubMedGoogle Scholar
  • 30 Lopez-Lazaro M. Dual role of hydrogen peroxide in cancer: Possible relevance to cancer chemoprevention and therapy. Cancer Lett 2007; 252: 1-8
    CrossrefPubMedGoogle Scholar
  • 31 Lopez-Lazaro M. A new view of carcinogenesis and an alternative approach to cancer therapy. Mol Med 2010; 16: 144-153
    PubMedGoogle Scholar
  • 32 Martin-Cordero C, Leon-Gonzalez AJ, Calderon-Montano JM, Burgos-Moron E, Lopez-Lazaro M. Pro-oxidant natural products as anticancer agents. Curr Drug Targets 2012; 13: 1006-1028
    CrossrefPubMedGoogle Scholar
  • 33 Newman RA, Yang P, Hittelman WN, Lu T, Ho DH, Ni D, Chan D, Vijjeswarapu M, Cartwright C, Dixon S, Felix E, Addington C. Oleandrin-mediated oxidative stress in human melanoma cells. J Exp Ther Oncol 2006; 5: 167-181
    PubMedGoogle Scholar
  • 34 Helleday T, Petermann E, Lundin C, Hodgson B, Sharma RA. DNA repair pathways as targets for cancer therapy. Nat Rev Cancer 2008; 8: 193-204
    CrossrefPubMedGoogle Scholar
  • 35 Lord CJ, Ashworth A. The DNA damage response and cancer therapy. Nature 2012; 481: 287-294
    CrossrefPubMedGoogle Scholar
  • 36 Postel-Vinay S, Vanhecke E, Olaussen KA, Lord CJ, Ashworth A, Soria JC. The potential of exploiting DNA-repair defects for optimizing lung cancer treatment. Nat Rev Clin Oncol 2012; 9: 144-155
    CrossrefPubMedGoogle Scholar
  • 37 Bielawski K, Winnicka K, Bielawska A. Inhibition of DNA topoisomerases I and II, and growth inhibition of breast cancer MCF-7 cells by ouabain, digoxin and proscillaridin A. Biol Pharm Bull 2006; 29: 1493-1497
    CrossrefPubMedGoogle Scholar
  • 38 Jornada LK, Valvassori SS, Arent CO, Leffa D, Damiani AA, Hainzenreder G, Ferreira CL, Moretti M, Andrade VM, Quevedo J. DNA damage after intracerebroventricular injection of ouabain in rats. Neurosci Lett 2010; 471: 6-9
    CrossrefPubMedGoogle Scholar
  • 39 Hiyoshi H, Abdelhady S, Segerstrom L, Sveinbjornsson B, Nuriya M, Lundgren TK, Desfrere L, Miyakawa A, Yasui M, Kogner P, Johnsen JI, Andang M, Uhlen P. Quiescence and gammaH2AX in neuroblastoma are regulated by ouabain/Na,K-ATPase. Br J Cancer 2012; 106: 1807-1815
    CrossrefPubMedGoogle Scholar
  • 40 Gupta RS, Chopra A, Stetsko DK. Cellular basis for the species differences in sensitivity to cardiac glycosides (digitalis). J Cell Physiol 1986; 127: 197-206
    CrossrefPubMedGoogle Scholar
  • 41 Paul RJ, Bauer M, Pease W. Vascular smooth muscle: aerobic glycolysis linked to sodium and potassium transport processes. Science 1979; 206: 1414-1416
    CrossrefPubMedGoogle Scholar
  • 42 James JH, Fang CH, Schrantz SJ, Hasselgren PO, Paul RJ, Fischer JE. Linkage of aerobic glycolysis to sodium-potassium transport in rat skeletal muscle. Implications for increased muscle lactate production in sepsis. J Clin Invest 1996; 98: 2388-2397
    CrossrefPubMedGoogle Scholar
  • 43 Ishikawa N, Oguri T, Isobe T, Fujitaka K, Kohno N. SGLT gene expression in primary lung cancers and their metastatic lesions. Jpn J Cancer Res 2001; 92: 874-879
    CrossrefPubMedGoogle Scholar
  • 44 Yang P, Menter DG, Cartwright C, Chan D, Dixon S, Suraokar M, Mendoza G, Llansa N, Newman RA. Oleandrin-mediated inhibition of human tumor cell proliferation: importance of Na,K-ATPase alpha subunits as drug targets. Mol Cancer Ther 2009; 8: 2319-2328
    CrossrefPubMedGoogle Scholar
  • 45 Mijatovic T, Roland I, Van Quaquebeke E, Nilsson B, Mathieu A, Van Vynckt F, Darro F, Blanco G, Facchini V, Kiss R. The alpha1 subunit of the sodium pump could represent a novel target to combat non-small cell lung cancers. J Pathol 2007; 212: 170-179
    CrossrefPubMedGoogle Scholar
  • 46 Yang P, Cartwright C, Efuet E, Hamilton SR, Wistuba II, Menter D, Addington C, Shureiqi I, Newman RA. Cellular location and expression of Na(+), K(+) -ATPase alpha subunits affect the anti-proliferative activity of oleandrin. Mol Carcinogen advance online publication 16 Oct 2012 DOI: 10.1002/mc.21968.
    PubMedGoogle Scholar
  • 47 Chou TC. Drug combination studies and their synergy quantification using the Chou-Talalay method. Cancer Res 2010; 70: 440-446
    CrossrefPubMedGoogle Scholar
  • 48 Kishimoto S, Kawazoe Y, Ikeno M, Saitoh M, Nakano Y, Nishi Y, Fukushima S, Takeuchi Y. Role of Na+, K+-ATPase alpha1 subunit in the intracellular accumulation of cisplatin. Cancer Chemother Pharmacol 2006; 57: 84-90
    CrossrefPubMedGoogle Scholar
  • 49 Mijatovic T, Dufrasne F, Kiss R. Cardiotonic steroids-mediated targeting of the Na(+)/K(+)-ATPase to combat chemoresistant cancers. Curr Med Chem 2012; 19: 627-646
    CrossrefPubMedGoogle Scholar