Anti-proliferative and angio-suppressive effect of Stoechospermum marginatum (C. Agardh) Kutzing extract using various experimental models

  • Vinayak, Rashmi (Biological Oceanography Division, National Institute of Oceanography) ;
  • Puttananjaiah, Shilpa (Department of Studies in Biotechnology, University of Mysore) ;
  • Chatterji, Anil (Biological Oceanography Division, National Institute of Oceanography) ;
  • Salimath, Bharati (Department of Studies in Biotechnology, University of Mysore)
  • Received : 2013.05.14
  • Accepted : 2014.02.09
  • Published : 2014.08.01


BACKGROUND/OBJECTIVES: Abundant consumption of seaweeds in the diet is epidemiologically linked to the reduction in risk of developing cancer. In larger cases, however, identification of particular seaweeds that are accountable for these effects is still lacking, hindering the recognition of competent dietary-based chemo preventive approaches. The aim of this research was to establish the antiproliferative potency and angiosuppressive mode of action of Stoechospermum marginatum seaweed methanolic extract using various experimental models. MATERIALS/METHODS: Among the 15 seaweeds screened for antiproliferative activity against Ehrlich ascites tumor (EAT) cell line, Stoechospermum marginatum extract (SME) was found to be the most promising. Therefore, it was further investigated for its anti-proliferative activity in-vitro against choriocarcinoma (BeWo) and non-transformed Human embryonic kidney (HEK 293) cells, and for its anti-migratory/tube formation activity against HUVEC cells in-vitro. Subsequently, the angiosuppressive activity of S. marginatum was established by inhibition of angiogenesis in in-vivo (peritoneal angiogenesis and chorioallantoic membrane assay) and ex-vivo (rat cornea assay) models. RESULTS: Most brown seaweed extracts inhibited the proliferation of EAT cells, while green and red seaweed extracts were much less effective. According to the results, SME selectively inhibited proliferation of BeWo cells in-vitro in a dose-dependent manner, but had a lesser effect on HEK 293 cells. SME also suppressed the migration and tube formation of HUVEC cells in-vitro. In addition, SME was able to suppress VEGF-induced angiogenesis in the chorio allantoic membrane, rat cornea, and tumor induced angiogenesis in the peritoneum of EAT bearing mice. A decrease in the microvessel density count and CD31 antigen staining of treated mice peritoneum provided further evidence of its angiosuppressive activity. CONCLUSIONS: Altogether, the data underline that VEGF mediated angiogenesis is the target for the angiosuppressive action of SME and could potentially be useful in cancer prevention or treatment involving stimulated angiogenesis.


  1. Nussenbaum F, Herman IM. Tumor angiogenesis: insights and innovations. J Oncol 2010;2010:132641.
  2. Gupta K, Zhang J. Angiogenesis: a curse or cure? Postgrad Med J 2005;81:236-42.
  3. Folkman J. Angiogenesis in cancer, vascular, rheumatoid and other disease. Nat Med 1995;1:27-31.
  4. Hanahan D, Folkman J. Patterns and emerging mechanisms of the angiogenic switch during tumorigenesis. Cell 1996;86:353-64.
  5. Folkman J, Cotran R. Relation of vascular proliferation to tumor growth. Int Rev Exp Pathol 1976;16:207-48.
  6. Tanghetti E, Ria R, Dell'Era P, Urbinati C, Rusnati M, Ennas MG, Presta M. Biological activity of substrate-bound basic fibroblast growth factor (FGF2): recruitment of FGF receptor-1 in endothelial cell adhesion contacts. Oncogene 2002;21:3889-97.
  7. Carmeliet P. Mechanisms of angiogenesis and arteriogenesis. Nat Med 2000;6:389-95.
  8. Keshet E, Ben-Sasson SA. Anticancer drug targets: approaching angiogenesis. J Clin Invest 1999;104:1497-501.
  9. Madhusudan S, Harris AL. Drug inhibition of angiogenesis. Curr Opin Pharmacol 2002;2:403-14.
  10. Marwick C. Natural compounds show antiangiogenic activity. J Natl Cancer Inst 2001;93:1685.
  11. Smit AJ. Medicinal and pharmaceutical uses of seaweed natural products: a review. J Appl Phycol 2004;16:245-62.
  12. Chandini SK, Ganesan P, Bhaskar N. In vitro antioxidant activities of three selected brown seaweeds of India. Food Chem 2008;107: 707-
  13. 13. Ganesan P, Kumar CS, Bhaskar N. Antioxidant properties of methanol extract and its solvent fractions obtained from selected Indian red seaweeds. Bioresour Technol 2008;99:2717-23.
  14. Kumar KS, Ganesan K, Subba Rao PV. Antioxidant potential of solvent extracts of Kappaphycus alvarezii (Doty) Doty - an edible seaweed. Food Chem 2008;107:289-95.
  15. Devi KP, Suganthy N, Kesika P, Pandian SK. Bioprotective properties of seaweeds: in vitro evaluation of antioxidant activity and antimicrobial activity against food borne bacteria in relation to polyphenolic content. BMC Complement Altern Med 2008;8:38.
  16. Mojzis J, Varinska L, Mojzisova G, Kostova I, Mirossay L. Antiangiogenic effects of flavonoids and chalcones. Pharmacol Res 2008;57:259-65.
  17. Sugawara T, Baskaran V, Tsuzuki W, Nagao A. Brown algae fucoxanthin is hydrolyzed to fucoxanthinol during absorption by Caco-2 human intestinal cells and mice. J Nutr 2002;132:946-51.
  18. Ye J, Li Y, Teruya K, Katakura Y, Ichikawa A, Eto H, Hosoi M, Hosoi M, Nishimoto S, Shirahata S. Enzyme-digested fucoidan extracts derived from seaweed Mozuku of Cladosiphon novae-caledoniae kylin inhibit invasion and angiogenesis of tumor cells. Cytotechnology 2005;47:117-26.
  19. Dias PF, Siqueira JM Jr, Maraschin M, Ferreira AG, Gagliardi AR, Ribeiro-do-Valle RM. A polysaccharide isolated from the brown seaweed Sargassum stenophyllum exerts antivasculogenic effects evidenced by modified morphogenesis. Microvasc Res 2008;75: 34-44.
  20. Guerra Dore CM, Faustino Alves MG, Santos ND, Cruz AK, Câmara RB, Castro AJ, Guimarães Alves L, Nader HB, Leite EL. Antiangiogenic activity and direct antitumor effect from a sulfated polysaccharide isolated from seaweed. Microvasc Res 2013;88:12-8.
  21. Matsubara K, Mori M, Matsumoto H, Hori K, Miyazawa K. Antiangiogenic properties of a sulfated galactan isolated from a marine green alga, Codium cylindricum. J Appl Phycol 2003;15:87-90.
  22. Matsubara K, Xue C, Zhao X, Mori M, Sugawara T, Hirata T. Effects of middle molecular weight fucoidans on in vitro and ex vivo angiogenesis of endothelial cells. Int J Mol Med 2005;15:695-9.
  23. Vinayak RC, Sabu AS, Chatterji A. Bio-evaluation of two red seaweeds for their cytotoxic and antioxidant activities in vitro. J Complement Integr Med 2010;7. doi: 10.2202/1553-3840.1444.
  24. Vinayak RC, Sabu AS, Chatterji A. Bio-prospecting of a few brown seaweeds for their cytotoxic and antioxidant activities. Evid Based Complement Alternat Med 2011;2011:673083.
  25. Vinayak RC, Sudha SA, Chatterji A. Bio-screening of a few green seaweeds from India for their cytotoxic and antioxidant potential. J Sci Food Agric 2011;91:2471-6.
  26. Naderali EK, Brown MJ, Pickavance LC, Wilding JP, Doyle PJ, Williams G. Dietary obesity in the rat induces endothelial dysfunction without causing insulin resistance: a possible role for triacylglycerols. Clin Sci (Lond) 2001;101:499-506.
  27. Liang CC, Park AY, Guan JL. In vitro scratch assay: a convenient and inexpensive method for analysis of cell migration in vitro. Nat Protoc 2007;2:329-33.
  28. Gururaj AE, Belakavadi M, Salimath BP. Antiangiogenic effects of butyric acid involve inhibition of VEGF/KDR gene expression and endothelial cell proliferation. Mol Cell Biochem 2003;243:107-12.
  29. Sarayba MA, Li L, Tungsiripat T, Liu NH, Sweet PM, Patel AJ, Osann KE, Chittiboyina A, Benson SC, Pershadsingh HA, Chuck RS. Inhibition of corneal neovascularization by a peroxisome proliferator- activated receptor-gamma ligand. Exp Eye Res 2005;80:435-42.
  30. Sheela ML, Ramakrishna MK, Salimath BP. Angiogenic and proliferative effects of the cytokine VEGF in Ehrlich ascites tumor cells is inhibited by Glycyrrhiza glabra. Int Immunopharmacol 2006;6:494-8.
  31. Deepak AV, Salimath BP. Antiangiogenic and proapoptotic activity of a novel glycoprotein from U. indica is mediated by NF-kappaB and Caspase activated DNase in ascites tumor model. Biochimie 2006;88:297-307.
  32. Klement G, Baruchel S, Rak J, Man S, Clark K, Hicklin DJ, Bohlen P, Kerbel RS. Continuous low-dose therapy with vinblastine and VEGF receptor-2 antibody induces sustained tumor regression without overt toxicity. J Clin Invest 2000;105:R15-24.
  33. Kerbel R, Folkman J. Clinical translation of angiogenesis inhibitors. Nat Rev Cancer 2002;2:727-39.
  34. Browder T, Butterfield CE, Kräling BM, Shi B, Marshall B, O'Reilly MS, Folkman J. Antiangiogenic scheduling of chemotherapy improves efficacy against experimental drug-resistant cancer. Cancer Res 2000;60:1878-86.
  35. Miller KD, Sweeney CJ, Sledge GW Jr. Redefining the target: chemotherapeutics as antiangiogenics. J Clin Oncol 2001;19:1195-206.
  36. Hanahan D, Bergers G, Bergsland E. Less is more, regularly: metronomic dosing of cytotoxic drugs can target tumor angiogenesis in mice. J Clin Invest 2000;105:1045-7.
  37. Kotnala S, Garg A, Chatterji A. Screening for the presence of antimicrobial activity in few Indian seaweeds. Pertanika J Trop Agric Sci 2009;32:69-75.
  38. Folkman J, Shing Y. Angiogenesis. J Biol Chem 1992;267:10931-4.
  39. Ribatti D, Nico B, Vacca A, Roncali L, Burri PH, Djonov V. Chorioallantoic membrane capillary bed: a useful target for studying angiogenesis and anti-angiogenesis in vivo. Anat Rec 2001;264:317-24.
  40. Borgstrom P, Hillan KJ, Sriramarao P, Ferrara N. Complete inhibition of angiogenesis and growth of microtumors by anti-vascular endothelial growth factor neutralizing antibody: novel concepts of angiostatic therapy from intravital videomicroscopy. Cancer Res 1996;56:4032-9.
  41. Kim KJ, Li B, Winer J, Armanini M, Gillett N, Phillips HS, Ferrara N. Inhibition of vascular endothelial growth factor-induced angiogenesis suppresses tumour growth in vivo. Nature 1993;362:841-4.
  42. Mesiano S, Ferrara N, Jaffe RB. Role of vascular endothelial growth factor in ovarian cancer: inhibition of ascites formation by immunoneutralization. Am J Pathol 1998;153:1249-56.
  43. Folkman J. What is the evidence that tumors are angiogenesis dependent? J Natl Cancer Inst 1990;82:4-6.
  44. Sithranga Boopathy N, Kathiresan K. Anticancer drugs from marine flora: an overview. J Oncol 2010;2010:214186.
  45. Birner P, Obermair A, Schindl M, Kowalski H, Breitenecker G, Oberhuber G. Selective immunohistochemical staining of blood and lymphatic vessels reveals independent prognostic influence of blood and lymphatic vessel invasion in early-stage cervical cancer. Clin Cancer Res 2001;7:93-7.
  46. Cai H, Harrison DG. Endothelial dysfunction in cardiovascular diseases: the role of oxidant stress. Circ Res 2000;87:840-4.
  47. Wang YD, Wu P, Mao JD, Huang H, Zhang F. Relationship between vascular invasion and microvessel density and micrometastasis. World J Gastroenterol 2007;13:6269-73.

Cited by

  1. Methanolic Extracts from Brown Seaweeds Dictyota cilliolata and Dictyota menstrualis Induce Apoptosis in Human Cervical Adenocarcinoma HeLa Cells vol.20, pp.4, 2015,