Planta Med 2006; 72(1): 20-27
DOI: 10.1055/s-2005-873167
Original Paper
© Georg Thieme Verlag KG Stuttgart · New York

Direct and Indirect Mechanism(s) of Antitumour Activity of Propolis and its Polyphenolic Compounds

Nada Oršolić1 , Ana Brbot Šaranović2 , Ivan Bašić1
  • 1Department of Animal Physiology, Faculty of Science, University of Zagreb, Zagreb, Croatia
  • 2Department of Chemistry, Veterinary Faculty, University of Zagreb, Zagreb, Croatia
Further Information

Publication History

Received: December 22, 2004

Accepted: June 20, 2005

Publication Date:
10 November 2005 (online)


The immunomodulatory actions of a water-soluble derivative of propolis (WSDP) and two components of propolis, caffeic acid (CA) and caffeic acid phenethyl ester (CAPE) were investigated. Oral administration (50 mg/kg) of WSDP, CA, and CAPE enhanced the weight and cellularity of the spleen (p < 0.05, p < 0.01) of treated mice. The response of spleen cells to polyclonal mitogens (PHA, Con A, PWM) was also increased in mice treated with WSDP as compared to control (p < 0.01); in contrast, the response of spleen cells of mice treated with CA were significantly suppressed (p < 0.001). The colony forming ability of HeLa cells plated on monolayers of macrophages was completely inhibited by peritoneal macrophages from mice receiving either WSDP, CAPE, or CA. Macrophages from treated mice also inhibited [3 H]TdR incorporation into HeLa cells in vitro. Testing for the possible presence of NO in the supernatants of 24 hours cultured macrophages activated with either compound revealed that the toxicity of these cells to HeLa cells was in part due to the production of NO. Tumour growth was suppressed by WSDP and its polyphenolic compounds given orally to mice. Local presence of CA, and CAPE in the tissue, caused a significant delay of tumour formation. Based on these results, we postulate that the antitumour activity of the test compounds includes pronounced immunomodulatory activity mainly due to the augmentation of non-specific antitumour resistance in mice via macrophage activation and the production of soluble factors by those cells which may interfere with either cells of the immune system or directly by tumour cells.


WSDP:water-soluble derivative of propolis

CA:caffeic acid

CAPE:caffeic acid phenethyl ester

NO:nitric oxide

NOS:nitric oxide syntase

[3 H]-TdR:[3 H]-thymidine


  • 1 Monacada S, Palmer R MJ, Higgs E A. Nitric oxide: physiology, pathophysiology, and pharmacology.  Pharmacol Rev. 1991;  43 109-42
  • 2 Lowenstein C J, Snyder S H. Nitric oxide, a novel biologic messenger.  Cell. 1992;  70 705-7
  • 3 Xie Q W, Nathan C. The high-output nitric oxide pathway: role and regulation.  J Leukoc Biol. 1992;  56 576-80
  • 4 Murad J M, Calvi S A, Soares A MVC, Bankova V, Sforcin J M. Effect of propolis from Brazil and Bulgaria on fungicidal activity of macrophages against Paracoccidioides brasiliensis .  J Ethnopharmacol. 2002;  79 304-31
  • 5 Tatefuji T, Izumi N, Ohta T, Arai S, Ikeda M, Kurimoto M. Isolation and identification of compounds from Brazilian propolis which enhance macrophage spreading and mobility.  Biol Pharm Bull. 1996;  19 966-70
  • 6 Dimov V, Ivanovska N, Manolova N, Bankova V, Nikolov N, Popov S. Immunodulatory action of propolis. Influence on anti-infectious protection and macrophage function.  Apidologie. 1991;  22 155-62
  • 7 Oršolić N, Bašić I. Immunomodulation by water-soluble derivative of propolis (WSDP) a factor of antitumor reactivity.  J Ethnopharmacol. 2003;  84 265-73
  • 8 Bašić I, Oršolić N, Tadić Z, Macedo Fereire Alcici N, Brbot Šaranović A, Bendelja K. et al . Antimetastatic effect of propolis, caffeic acid phenethyl ester and caffeic acid on mammary carcinoma of CBA mouse.  Rio de Janeiro: 17th International Cancer Congress. 1998;  1 63-75
  • 9 Dimov V, Ivanovska N, Bankova V, Popov S. Immunodulatory action of propolis. IV. Prophylactic activity against Gram-negative infections and adjuvant effect of the water-soluble derivate.  Vaccine. 1992;  10 817-23
  • 10 Meltzer M S, Tucker R W, Sanford K K, Leonard E J. Interaction of Bcg-activated macrophages with neoplastic and non-neoplastic cell line in vitro: quantitation of the cytotoxic reaction by release of tritiated thymidine from prelabeled target cells.  J Natl Cancer Inst. 1975;  45 1177-84
  • 11 Stuehr D J, Nathan C F. Nitric oxide: a macrophage product responsible for cytostasis and respiratory inhibition in tumor target cells.  J Exp Med. 1989;  169 543-55
  • 12 Elgert K D, Allleva G D, Mullins D W. Tumor-induced immune disfunction: the macrophage connection.  J Leukoc Biol. 1998;  64 275-90
  • 13 Nikolov N, Marekov N, Bankova V, Popov S, Ignatova R, Vladimirova I. Method for the preparation of water-soluble derivatives of propolis.  Bulg Patent Appl. 1987;  79 903/28 05
  • 14 Grunberger D, Banerjee R, Eisinger K, Oltz Efros E HL, Caldwell M, Estevez V. et al . Preferential cytotoxicity on tumor cells of caffeic acid phenethyl ester isolated from propolis.  Experientia. 1988;  44 230-2
  • 15 Bašić I, Varga E. Immunogenicity of mammary carcinoma and a fibrosarcoma of CBA mice.  Period Biol. 1979;  81 335-7
  • 16 Yoshikawa T, Kokura S, Tainaka K, Naito Y, Kondo M. A novel cancer therapy based on oxygen radicals.  Cancer Res. 1999;  55 1617-20
  • 17 Plowman J, Dykes D J, Hollingshead M, Simpson-Herren L, Alley M C. Human tumor xenograft models in NCI drug development. In: Teicher B, editor Anticancer drug development guide: preclinical screening, clinical trials, and approval. Totowa; Humana 1995: p 101
  • 18 Kimoto T, Arai S, Kohguchi M, Aga M, Nomura Y, Micallef M J. et al . Apoptosis and suppression of tumor growth by artepillin C extracted from Brazilian propolis.  Cancer Detect Prev. 1998;  22 506-15
  • 19 Hayashi A, Gillen A C, Lott J R. Effects of daily oral administration of quercetin chalcone and modified citrus pectin on implanted colon-25 tumor growth in Balb-c mice.  Altern Med Rev. 2000;  5 546-52
  • 20 Scheller S, Krol W, Swiacik J, Owczarek S, Gabrys J, Shani J. Antitumoral property of ethanolic extract of propolis in mice-bearing Ehrlich carcinoma, as compared to bleomycin.  Z Naturforsch [C]. 1989;  44c 1063-5
  • 21 Matsuno T. A new clerodane diterpenoid isolated from propolis.  Z Naturforsch [C]. 1995;  50 93-7
  • 22 Sotomayor E M, Dinapoli M R, Calderon C, Colsky A, Fu YX Lopez D M. Decreased macrophage-mediated cytotoxicity in mammary-tumor-bearing mice is related to alteration of nitric oxide production and/or release.  Int J Cancer. 1995;  60 660-7
  • 23 Kono K, Salazaronfray F, Petersson M, Hansson J, Masucci G, Wasserman K. et al . Hydrogen peroxide secreted by tumor-derived macrophages down-modulates signal-transducing zeta molecules and inhibits tumor-specific T cell- and natural killer cell-mediated cytotoxicity.  Eur J Immunol. 1996;  26 1308-13
  • 24 Ivanovska M, Stefanova Z, Valeva V, Neychev H. Immunodulatory action of propolis: VII. A comparative study on cinnamic and caffeic acid lysine derivatives.  Biol Immun. 1993;  46 115-7
  • 25 Orsi R O, Funari S RC, Soares A MVC, Calvi S A, Oliveira S L, Sforcin J M. et al . Immunomodulatory action of propolis on macrophage activation.  J Venom Anim Toxins. 2000;  6 1-11
  • 26 Chan W S, Wen P C, Chiang H C. Structure-activity relationship of caffeic acid analogues on xanthine oxidase.  Anticancer Res. 1995;  15 703-7
  • 27 Monks T J, Lau S S. Biological reactivity of polyphenolic-glutathione conjugates.  Chem Res Toxicol. 1997;  10 1296-313
  • 28 Nathan C F. Secretory products of macrophages.  J Clin Invest. 1987;  79 319-26
  • 29 Oršolić N, Bašić I. Immunostimulatory and antitumor activity of a water-soluble derivative of propolis (WSDP).  Period Biol. 2001;  103 255-61

Nada Oršolić

Department of Animal Physiology

Faculty of Science

University of Zagreb

10000 Zagreb

Rooseveltov trg 6


Phone: +385-1-4826-266

Fax: +385-1-4826-269/280/313