Significant anticancer activity of a venom fraction derived from the Persian Gulf sea anemone, Stichodactyla haddoni

Document Type : Research article


1 Venom and Biotherapeutics Molecules Lab., Medical Biotechnology Dept., Biotechnology Research Center, Pasteur Institute of Iran. Tehran, Iran.

2 Iranian Fisheries Science Research Institute, Agricultural Research, Education and Extension Organization

3 Department of Immunology, Faculty of Medicine, Shahid Beheshti University of Medical Sciences. Tehran, Iran.


Chemotherapy is still one of the main therapeutic regimens in cancer patients but its toxicity is a hard challenge for every patient yet. One of the available solutions is tracing for non-toxic anticancer agents from natural resources. Numerous proteins and peptides in the venom of sea anemones are potentially useful agents with pharmacological properties. Concerning to significance of this issue, the current study was aimed to finding a non-toxic anticancer fraction from the venom of the Persian Gulf sea anemone, Stichodactyla haddoni. Anticancer and hemolytic activity of crude venom was evaluated and followed by fractionation using RP-HPLC. Breast, Brain, and Colon cancer cell lines were selected to assessment of anticancer activity and toxicity. EC50 of crude venom on the abovementioned cancer cell lines were as 3.12, 4.32, and 12.5 µg, respectively. According to results obtained by paired sample t-test and comparison of toxicity of the fractions in normal cell line, F10, designated as hadonin, was determined as the candidate anti-cancer fraction. The non-toxic dose of F10 was 20 ng in which showed respectively 66, 29, and 7 anticancer activities on breast, brain, and colon cancer cell lines. According to results, anticancer activity of hadonin is of high pharmaceutical value to follow its therapeutic potency in animal model. Our result is the first report of an anticancer ca. 17.5 kDa protein in the Persian Gulf sea anemone with reasonable activity at nanogram level against three kinds of cancer cells with no toxicity on normal cells.


  1. Kirthi C, Afzal A, Reddy M, Ali SA, Yerramilli C and Sharma S. A study on the adverse effects of anticancer drugs in an oncology center of a tertiary care hospital. J. Pharmacy. Pharmaceut. Sci. (2014) 6: 580-3.
  2. Üskent N, Demirbas S, Turken O, Yildirim Ş, Tecimer C, Kandemir G and Yaylaci M. Survival from the precocious brain metastasis of the colon cancer. J. Cancer (2003) 33: 154-7.
  3. Aslam MS, Naveed S, Ahmed A, Abbas Z, Gull I and Athar MA. Side effects of chemotherapy in cancer patients and evaluation of patients opinion about starvation based differential chemotherapy. Cancer Therapy. (2014) 5: 817-22.
  4. Malve H. Exploring the ocean for new drug developments: Marine pharmacology. J Pharm. Bioallied. Sci. (2016) 8: 83–91.
  5. Cheung RCF, Ng TB, Wong JH. Marine Peptides: Bioactivities and Applications. Drugs (2015) 13: 4006-4043.
  6. Michael TDC, Clinton GLV. Recent Advances in Drug Discovery from South African Marine Invertebrates. Mar Drugs (2015) 13: 6366-83.
  7. Hu Y, Chen J, Hu G, Yu J, Zhu X, Lin Y, Chen S and Yuan J. Statistical Research on the Bioactivity of New Marine Natural Products Discovered during the 28 Years from 1985 to 2012. Drugs (2015) 13: 202-21.
  8. Harvey A L. Toxins and drug discovery. Toxicon (2014) 92: 193-200.
  9. Martins RD, Alves RS, Martins AMC, Barbosa PSF, Evangelista JSAM, Evangelista JJF, Ximenes RM, Toyama MH,Toyama DO, Souza AJF, Orts DJB, Marangoni S, de Menezes DB, Fonteles MC and Monteiro HSA. Purification and characterization of the biological effects of phospholipase A2 from sea anemone Bunodosoma caissarum. Toxicon (2009) 54: 413–20.
  10. Mariottini GL and Pane L. Cytotoxic and cytolytic cnidarian venoms. A review on health implications and possible therapeutic applications. Toxins (2014) 6: 108-151.
  11. Newman DJ and Cragg MG. Marine natural products and related compounds in clinical and advanced preclinical trials. Anti-Cancer Agents Med. Chem. (2013) 13: 603-31.
  12. Petit K., Biard J.F. Marine Natural products and related compounds as anticancer agents: an overview of their clinical status. Anti-Cancer Agents Med. Chem. (2013) 13: 603-31.
  13. Essack M, Bajic VB, Archer JA. Conotoxins that confer therapeutic possibilities. Mar Drugs (2012) 10: 1244-65.
  14. Chi V, Pennington MW, Norton RS, Tarcha EJ, Londono LM, Sims-Fahey B, Upadhyay SK, Lakey JT, Iadonato S, Wulff H, Beeton C and Chandy KG. Development of a sea anemone toxin as an immunomodulator for therapy of autoimmune diseases. Toxicon (2012) 59: 529-46.
  15. Molinski TF, Dalisay DS, Lievens SL and Saludes JP. Drug development from marine natural products. Nat .Rev. Drug Discov. (2009) 8: 69-85.
  16. Leal M, Sapra P, Hurvitz SA, Senter P, Wahl A, Schutten M, Shah DK, Haddish-Berhane N and Kabbarah O. Antibody-drug conjugates: An emerging modality for thetreatment of cancer. NY Acad Sci (2014) 1321: 41–54.
  17. Rinehart KL, Holt TG, Fregeau NL, Stroh JG, Keifer PA, Sun F, Li LH, Martin DG. Ecteinascidins 729, 743, 745, 759A, 759B, and 770: potent antitumor agents from the Caribbean tunicate Ecteinascidia turbinate. Org. Chem. (1990) 55: 4512–5.
  18. The Complete Drug Reference (database on the internet). Cytarabine. Thomson MICROMEDEX. (2009).
  19. Lichtman MA. A historical perspective on the development of the cytarabine (7 days) and daunorubicin (3 days) treatment regimen for acute myelogenous leukemia: 2013 the 40th anniversary of 7 + 3. Blood Cells Mol Dis. (2013) 50: 119–30.
  20. Shen W, Kim JS, Kish PE, Zhang J, Mitchell S, Gentry BG, Breitenbach JM, Drach JC, Hilfinger J. Design and synthesis of Vidarabine prodrugs as antiviral agents. Med Chem Lett. (2009) 19: 792–6.                                                                   
  21. Cuadrado A, Garcia-Fernandez LF, Gonzalez L, Gonzalez L, Suarez Y, Losada A, Alcaide V, Martinez T, Fernandez-Sousa JM, Sanchez-Puelles JM and Munoz A. Aplidin induces apoptosis in human cancer cells via glutathione depletion and sustained activation of the epidermal growth factor receptor, Src, JNK, and p38 MAPK. Biol. Chem. (2003) 278: 241–50.
  22. Sudek S, Lopanik NB, Waggoner LE, Hildebrand M, Anderson C, Liu H, Patel A, Sherman DH, Haygood MG. Identification of the putative bryostatin polyketide synthase gene cluster from ―Candidatus Endobugula sertula‖, the uncultivated microbial symbiont of the marine bryozoan Bugula neritina. Nat. Prod. (2007) 70: 67–74.
  23. Talpir R, BenayahuY, Kashman L, Pannell L, Schleyer M. Hemiasterlin and geodiamolide TA: two new cytotoxic peptides from the marine sponge Hemiasterella minor. Tetra Lett. (1994) 35: 4453–6.
  24. LingYH, Aracil M, Jimeno J, Perez-Soler R and Zou1 Y. Molecular pharmacodynamics of PM02734 (elisidepsin) as single agent and in combination with erlotinib; synergistic activity in human non-small cell lung cancer cell lines and xenograft models. J. Cancer (2009) 45: 1855–64.
  25. Ramezanpour M, Burke da Silva, K. Sanderson BJS. Differential susceptibilities of human lung, breast and skin cancer cell lines to killing by five sea anemone venoms. Venom. Anim. Toxins: incl Trop. Dis. (2012) 18: 157-63.
  26. Monroy-Estrada H, Chirino Y, Soria-Mercado IE, Sánchez-Rodríguez J. Toxins from the Caribbean Sea anemone Bunodeopsis globulifera increase cisplatin-induced cytotoxicity of lung adenocarcinoma cells. J Venom Anim Toxins: incl Trop. Dis. (2013) 19: 12
  27. Soletti RC, de Faria GP, Vernal J, Terenzi H, Anderluh G, Borges HL, Moura-Neto V, Gabilan NH. Potentiation of anticancer-drug cytotoxicity by sea anemone pore-forming proteins in human glioblastoma cells. Antican Drugs (2008) 19: 517–25.
  28. Anderluh G., Maček P. Cytolytic peptide and protein toxins from sea anemones (Anthozoa:Actiniaria). Toxicon (2002) 40: 111–24.
  29. Maček P, Zecchini M, Stanek K and Menestrina G. Effect of membrane partitioned n-alcohols and fatty acids on pore-forming activity of a sea anemone toxin. Biophys. J. (1997) 25: 155–62.
  30. Tejuca M, Dalla Serra M, Potrich C, Alvarez C and Menestrina G. Sizing the radius of the pore formed in erythrocytes and lipid vesicles by the toxin Sticholysin I from the sea anemone Stichodactyla helianthus. Membr. Biol. (2001) 83: 125–35.
  31. Tejuca M, Anderluh G and Dalla Serra M. Sea anemone cytolysins as toxic components fo immnunotoxins. Toxicon (2009) 54: 1206–14.
  32. Tejuca M, Pérez-Barzaga V, Pazos F, Álvarez C, Lanio ME. Construction of sea anemone cytolysin-based immunotoxins for selective killing of cancer cells. Rev Cub Fisica. (2009) 26: 15-22.
  33. Fedorov S, Dyshlovoy S, Monastyrnaya M, Shubina L, Leychenko E, Kozlovskaya E, Jin JO, Kwak JY, Bode AM, Dong Z and Stonika V. The anticancer effects of actinoporin RTX-A from the sea anemone Heteractis crispa (=Radianthus macrodactylus). Toxicon (2010) 55: 811–7.
  34. Jiang X, Chen H, Yang W, Liu Y, Liu W, Wei J, Tu H, Xie X, Wang L and Xu A. Functional expression and characterization of an acidic actinoporin from sea anemone Sagartia rosea. Biophys. Res. Commun. (2003) 312: 562–70.
  35. Yan L, Herrington J, Goldberg E, Dulski PM, Bugianesi RM, Slaughter RS, Banerjee P, Brochu RM, Priest BT, Kaczorowski GJ, Rudy B and Garcia ML. Stichodactyla helianthus peptide, a pharmacological tool for studying Kv3.2 channels. Pharmacol. (2005) 67: 1513–21.
  36. Frazão B, Vasconcelos V and Antunes A. Sea Anemone (Cnidaria, Anthozoa, Actiniaria) Toxins. An Overview. Drugs (2012) 10: 1812-51.
  37. Kem WR. Pennington MW, Norton RS. Sea anemone toxins as templates for the design of immunosuppressant drugs. Drug Discov. Des. (1999) 15/16: 111-29.
  38. Laemmli U.K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature (1970) 227: 680-5.
  39. Memar B, Jamili S, Shahbazzadeh D and Pooshang Bagheri K. The first report on coagulation and phospholipase A2 activities of Persian Gulf lionfish, Pterois russelli, an Iranian venomous fish. Toxicon (2016) 113: 25-31.
  40. Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. Immunol. Methods (1983) 65: 55-63.
  41. Subramanian B, Sangappellai T, Rajak CR, Diraviam B. Pharmacological and biomedical properties of sea anemones Paracondactylis indicus, Paracondactylis sinensis, Heteractis magnifica and Stichodactyla haddoni from East coast of India. Asian Pacific J Trop Med. (2011) 722-726.
  42. Sudharsan S, Seedevi P, Kanagarajan U, Dalvi SR, Guptha S, Poojary N, Shanmugam V, Srinivasan A and Shanmugam A. Analgesic and neuromodulatory effects of sea anemone Stichodactyla mertensii (Brandt, 1835) methanolic extract from southeast coast of India. Afric J. Pharm. Pharmacol. (2013) 7: 2180-200.
  43. Marino A, Morabito R and La Spada G. Factors altering the haemolytic power of crude venom from mutabilis (Anthozoa) nematocysts. Comp Biochem Physiol. A. (2009) 152: 418–22.
  44. Cline EI, Wiebe LI, Young JD and Samuel J. Toxic effects of the novel protein UpI from the sea anemone Urticina piscivora. Pharmacol Res. (1995) 32: 309–14.
  45. Ravindran SV, Kannan L and Venkateshvaran K. Biological activity of sea anemons proteins:I Toxicity and histopatology. Indian journal of Experimental Biology. (2010) 47: 1225-1232.
  46. Avila AD, Mateo de Acosta C and Lage A. A new immunotoxin built by linking a hemolytic toxin to a monoclonal antibody specific for immature T lymphocytes. Int. J. Cancer (1988) 42: 568–71.
  47. Batista U, Macek P and Sedmak B. The cytotoxic and cytolytic activity of Equinatoxin II from the sea anemone Actinia equina. Cell Biol. Int. Rep. (1990) 14: 1013–24.