Investigational New Drugs

, Volume 30, Issue 6, pp 2187–2200 | Cite as

Quinones and halogenated monoterpenes of algal origin show anti-proliferative effects against breast cancer cells in vitro

  • Jo-Anne de la Mare
  • Jessica C. Lawson
  • Maynard T. Chiwakata
  • Denzil R. Beukes
  • Adrienne L. EdkinsEmail author
  • Gregory L. Blatch


Red and brown algae have been shown to produce a variety of compounds with chemotherapeutic potential. A recent report described the isolation of a range of novel polyhalogenated monoterpene compounds from the red algae Plocamium corallorhiza and Plocamium cornutum collected off the coast of South Africa, together with the previously described tetraprenylquinone, sargaquinoic acid (SQA), from the brown algae Sargassum heterophyllum. In our study, the algal compounds were screened for anti-proliferative activity against metastatic MDA-MB-231 breast cancer cells revealing that a number of compounds displayed anti-cancer activity with IC50 values in the micromolar range. A subset of the compounds was tested for differential toxicity in the MCF-7/MCF12A system and five of these, including sargaquinoic acid, were found to be at least three times more toxic to the breast cancer than the non-malignant cell line. SQA was further analysed in terms of its mechanism of cytotoxicity in MDA-MB-231 cells. The ability to initiate apoptosis was distinguished from the induction of an inflammatory necrotic response via flow cytometry with propidium iodide and Hoescht staining, confocal microscopy with Annexin V and propidium iodide staining as well as the PARP cleavage assay. We report that SQA induced apoptosis while a polyhalogenated monoterpene RU015 induced necrosis in metastatic breast cancer cells in vitro. Furthermore, we demonstrated that apoptosis induction by SQA occurs via caspase-3, -6, -8, -9 and -13 and was associated with down-regulation of Bcl-2. In addition, cell cycle analyses revealed that the compound causes G1 arrest in MDA-MB-231 cells.


Marine natural products Apoptosis Breast cancer Halogens Quinones 



This research was supported by grants from the Cancer Research Initiative of South Africa (CARISA) and the South African National Research Foundation (NRF). PhD Scholarships from Rhodes University (RU), the NRF and RU Henderson Foundation (JdlM) and Masters scholarships from the RU Henderson foundation, the German Academic Exchange Service (DAAD) and the Ernst and Ethel Eriksen trust (JCL) are hereby gratefully acknowledged.

Conflict of interest

The authors declare that they have no conflict of interest

Supplementary material

10637_2011_9788_MOESM1_ESM.jpg (239 kb)
Fig. S1 Kinetic study of the anti-proliferative effect of sargaquinoic acid against MDA-MB-231 cells. Cells were treated with 10, 40, 60, 80 and 100 μM of sargaquinoic acid over 24, 48, 72 or 96 h and their proliferative ability assessed by MTT assay. Percentage survival values were calculated relative to a DMSO vehicle-treated control. (JPEG 238 kb)
10637_2011_9788_MOESM2_ESM.jpg (264 kb)
Fig. S2 Measurement of apoptotic cell death by flow cytometry in MDA-MB-231 cells. Treatment was with DMSO (vehicle control), paclitaxel or RU015 for 15 h, followed by staining with Hoescht 33342 and/or Propidium iodide (PI) and analysis by flow cytometry (a) Gating carried out on a forward- and side-scatter representation of the unstained vehicle-treated sample and copied to all subsequent samples. Assessment of degree of apoptosis upon treatment with a range of concentrations of either (b) paclitaxel or (c) RU015. (JPEG 263 kb)
10637_2011_9788_MOESM3_ESM.jpg (178 kb)
Fig. S3 Cell cycle analysis by flow cytometry with propidium iodide staining. (a) Doublet discrimination to prevent false positives due to cell aggregates. Gating was carried out on an area (A) versus width (W) representation of the propidium iodide (PI)-stained DMSO vehicle-treated sample and copied to all subsequent samples. (b) Paclitaxel induces a G2-M phase arrest in MDA-MB-231 cells. Treatment was with either vehicle control (DMSO) or 50 nM paclitaxel (Ptx) for 16 h. Population gating was carried out according to the DMSO-treated control. (JPEG 177 kb)
10637_2011_9788_MOESM4_ESM.doc (28 kb)
Table S1 Pharmacological profiling of selected marine algal compounds using the Rule of 5 (Ro5) (DOC 28 kb)


  1. 1.
    World Health Statistics (2008) Part 1: ten highlights in health statistics, breast cancer: mortality and screening. World Health Organisation, Geneva, pp 21–23Google Scholar
  2. 2.
    Peto R, Boreham J, Clarke M, Davies C, Beral V (2000) UK and USA breast cancer deaths down 25% in year 2000 at ages 20–69 years. Lancet 355:1822–1822PubMedCrossRefGoogle Scholar
  3. 3.
    Howe HL, Wingo PA, Thun MJ, Ries LA, Rosenberg HM, Feigal EG, Edwards BK (2001) Annual report to the nation on the status of cancer (1973 through 1998), featuring cancers with recent increasing trends. J Natl Cancer Inst 93:824–842PubMedCrossRefGoogle Scholar
  4. 4.
    Zhou J, Zhang H, Gu P, Bai J, Margolick M, Zhang Y (2008) NF-κB pathway inhibitors preferentially inhibit breast cancer stem-like cells. Breast Canc Res Treat 111:419–427CrossRefGoogle Scholar
  5. 5.
    Jin Z, El-Deiry WS (2005) Overview of cell death signaling pathways. Canc Biol Ther 4(2):139–163CrossRefGoogle Scholar
  6. 6.
    Iannolo G, Conticello C, Memeo L, De Maria R (2008) Apoptosis in normal and cancer cells. Crit Rev Oncol Hematol 66:42–51PubMedCrossRefGoogle Scholar
  7. 7.
    Fink SL, Cookson BT (2005) Apoptosis, pyroptosis, and necrosis: mechanistic description of dead and dying eukaryotic cells. Infect Immun 73:1907–1916PubMedCrossRefGoogle Scholar
  8. 8.
    Klener P Jr, Andera L, Klener P, Necas E, Zivny J (2006) Cell death signalling pathways in the pathogenesis and therapy of haematologic malignancies: overview of apoptotic pathways. J Folia Biol (Praha) 52:34–44Google Scholar
  9. 9.
    Leist M, Jaattela M (2001) Four deaths and a funeral: from caspases to alternative mechanisms. Nat Rev Mol Cell Biol 2:589–98PubMedCrossRefGoogle Scholar
  10. 10.
    Nagata S (1997) Apoptosis by death factor. Cell 88:355–365PubMedCrossRefGoogle Scholar
  11. 11.
    Kroemer G, Reed JC (2000) Mitochondrial control of cell death. Nat Med 6:513–519PubMedCrossRefGoogle Scholar
  12. 12.
    Kerr JF, Wyllie AH, Currie AR (1972) Apoptosis: a basic biological phenomenon with wide-ranging implications in tissue kinetics. Br J Cancer 26:239–57PubMedCrossRefGoogle Scholar
  13. 13.
    Debatin K-M, Krammer PH (2004) Death receptors in chemotherapy and cancer. Oncogene 23:2950–2966PubMedCrossRefGoogle Scholar
  14. 14.
    Boulares AH, Yakovlev AG, Ivanova V, Stoica BA, Wang G, Iyer S, Smulson M (1999) Role of poly(ADP-ribose) polymerase (PARP) cleavage in apoptosis. Caspase 3-resistant PARP mutant increases rates of apoptosis in transfected cells. J Biol Chem 274:22932–22940PubMedCrossRefGoogle Scholar
  15. 15.
    Carté BK (1996) Biomedical potential of marine natural products. Bioscience 46(4):271–286CrossRefGoogle Scholar
  16. 16.
    Jha RK, Zi-rong X (2004) Biomedical compounds from marine organisms. Mar Drugs 2:123–146CrossRefGoogle Scholar
  17. 17.
    König GM, Wright AD, Franzblau SG (2000) Assessment of antimycobacterial activity of a series of mainly marine derived natural products. Planta Medica 66:337–342PubMedCrossRefGoogle Scholar
  18. 18.
    Blunt JW, Copp BR, Munro MHG, Northcote PT, Prinsep MR (2003) Marine natural products. Nat Prod Rep 20:1–48PubMedCrossRefGoogle Scholar
  19. 19.
    Knott MG, Mkwananzi HB, Arendse CE, Hendricks DT, Bolton JJ, Beukes DR (2005) Plocoralides A-C, polyhalogenated monoterpenes from the marine alga Plocamium corallorhiza. Phytochem 66:1108–1112CrossRefGoogle Scholar
  20. 20.
    Fuller RW, Cardellina JH II, Kato Y, Brinen LS, Clardy J, Snader KM, Boyd MR (1992) A pentahalogenated monoterpene from the red alga Portieria hornemannii produces a novel cytotoxicity profile against a diverse panel of human tumor cell lines. J Med Chem 35:3007–3011PubMedCrossRefGoogle Scholar
  21. 21.
    Mann MGA, Mkwananzi HB, Antunes EM, Whibley CE, Hendricks DT, Bolton JJ, Beukes DR (2007) Halogenated monoterpene aldehydes from the South African marine alga Plocomium corallorhiza. J Nat Prod 70:596–599PubMedCrossRefGoogle Scholar
  22. 22.
    Afolayan AF, Bolton JJ, Lategan CA, Smith PJ, Beukes DR (2008) Fucoxanthin, tetraprenylated toluquinone and toluhydroquinone metabolites from Sargassum heterophyllum inhibit the in vitro growth of the malaria parasite Plasmodium falciparum. Z Naturforsch 63c:848–852Google Scholar
  23. 23.
    Hur S, Lee H, Kim Y, Lee B-H, Shin J, Kim TY (2008) Sargaquinoic acid and sargachromenal, extracts of Sargassum sagamianum induce apoptosis in HaCaT cells and mice skin: Its potentiation of UVB-induced apoptosis. Eur J Pharmacol 582:1–11PubMedCrossRefGoogle Scholar
  24. 24.
    Warleta F, Campos M, Allouche Y, Sanchez-Quesada C, Ruiz-Mora J, Beltran G, Gaforio JJ (2010) Squalene protects against oxidative DNA damage in MCF10A human mammary epithelial cells but not in MCF-7 and MDA-MB-231 human breast cancer cells. Food Chem Toxicol 48:1092–1100PubMedCrossRefGoogle Scholar
  25. 25.
    Lipinski CA, Lombardo F, Dominy BW, Feeney PJ (2001) Experimental and computational approaches to estimate solubility and permeability in drug discovery and development settings. Adv Drug Deliv Rev 46:3–26PubMedCrossRefGoogle Scholar
  26. 26.
    Chiosis G (2003) Targeting chaperones in transformed systems–a focus on Hsp90 and cancer. Expert Opin Ther Targets 10:37–50CrossRefGoogle Scholar
  27. 27.
    Chaitanya GV, Babu PP (2009) Differential PARP cleavage: an indication of heterogeneous forms of cell death and involvement of multiple proteases in the infarct of focal cerebral ischemia in rat. Cell Mol Neurobiol 29:563–573PubMedCrossRefGoogle Scholar
  28. 28.
    Tzu-Hao W, Hsin-Shih W, Yung-Kwei S (2000) Paclitaxel induced cell death. Cancer 88:2619–2628CrossRefGoogle Scholar
  29. 29.
    Choi BW, Park SH, Kim ES, Shin J, Roh SS, Shin HC, Lee BH (2007) Anticholinesterase activity of plastoquinones from Sargassum sagamianum: lead compounds for Alzheimer’s disease therapy. Phytother Res 21:423–426PubMedCrossRefGoogle Scholar
  30. 30.
    Hosokawa M, Wanezaki S, Miyauchi K, Kurihara H, Kohno H, Kawabata J, Odashima S, Takahashi K (1999) Apoptosis-inducing effect of fucoxanthin on human leukemia cell line HL-60. Food Sci Technol Res 5(3):243–246CrossRefGoogle Scholar
  31. 31.
    Carr C, Ng J, Wigmore T (2008) The side effects of chemotherapeutic agents. Cur Anaes Crit Care 19:70–79CrossRefGoogle Scholar
  32. 32.
    Dwight ES, Lawrence WD, Carl C, Nanci LW, Hangming R, Gunter D (1997) Paclitaxel-induced apoptosis in MCF-7 breast-cancer cells. Int J Cancer 70:214–220CrossRefGoogle Scholar
  33. 33.
    Bachur NR, Gordon SL, Gee MV (1978) A general mechanism for microsomal activation of quinone anticancer agents to free radicals. Cancer Res 38:1745–1750PubMedGoogle Scholar
  34. 34.
    Hoyt MT, Palchaudhuri R, Hergenrother PJ (2011) Cribrostatin 6 induces death in cancer cells through a reactive oxygen species (ROS)-mediated mechanism. Investig New Drugs 29:562–573CrossRefGoogle Scholar
  35. 35.
    Bertram S (2000) The molecular biology of cancer. Mol Asp Med 21:167–223CrossRefGoogle Scholar
  36. 36.
    Johnstone RW, Ruefli AA, Lowe SW (2002) Apoptosis: a link between cancer genetics and chemotherapy. Cell 108:153–164PubMedCrossRefGoogle Scholar
  37. 37.
    Shao Y, Gao Z, Marks PA, Jiang X (2004) Apoptotic and autophagic cell death induced by histone deacetylase inhibitors. Proc Natl Acad Sci USA 101:18030–18035PubMedCrossRefGoogle Scholar
  38. 38.
    Kamei Y, Tsang CK (2003) Sargaquinoic acid promotes neurite outgrowth via protein kinase A and MAP kinases-mediated signaling pathways in PC12D cells. Int J Dev Neurosci 21:255–262PubMedCrossRefGoogle Scholar
  39. 39.
    Dive C, Gregory CD, Phipps DJ, Evans DL, Milner AE, Wyllie AH (1992) Analysis and discrimination of necrosis and apoptosis (programmed cell death) by multiparameter flow cytometry. Biochim Biophys Acta 1133:275–285PubMedCrossRefGoogle Scholar
  40. 40.
    Soldani C, Scovassi AI (2002) Poly(ADP-ribose) polymerase-1 cleavage during apoptosis: an update. Apoptosis 7:321–328PubMedCrossRefGoogle Scholar
  41. 41.
    Jacobson MD, Raff MC (1994) Programmed cell death and Bcl-2 protection in very low oxygen. Nature 374:814–816CrossRefGoogle Scholar
  42. 42.
    de Ines C, Argandona VH, Rovirosa J, San-Martin A, Diaz-Marrero AR, Cueto M, Gonzalez-Coloma A (2004) Cytotoxic activity of halogenated monoterpenes from Plocamium cartilagineum. Z Naturforsch 59:339–344Google Scholar

Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Jo-Anne de la Mare
    • 1
  • Jessica C. Lawson
    • 1
  • Maynard T. Chiwakata
    • 2
  • Denzil R. Beukes
    • 2
  • Adrienne L. Edkins
    • 1
    Email author
  • Gregory L. Blatch
    • 1
    • 3
  1. 1.The Biomedical Biotechnology Research Unit (BioBRU), Department of Biochemistry, Microbiology and BiotechnologyRhodes UniversityGrahamstownSouth Africa
  2. 2.Division of Pharmaceutical Chemistry, Faculty of PharmacyRhodes UniversityGrahamstownSouth Africa
  3. 3.School of Biomedical and Health Sciences, Faculty of Health, Engineering and ScienceVictoria UniversityMelbourneAustralia

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