Advertisement

Molecular and Cellular Biochemistry

, Volume 187, Issue 1–2, pp 191–199 | Cite as

The compound 14-keto-stypodiol diacetate from the algae Stypopodium flabelliforme inhibits microtubules and cell proliferation in DU-145 human prostatic cells

  • María S. Depix
  • Jorge Martínez
  • Francisco Santibañez
  • Juana Rovirosa
  • Aurelio San Martín
  • Ricardo B. Maccioni
Article

Abstract

We investigated the effects of the drug 14-keto-stypodiol diacetate (SDA) extracted from the seaweed product Stypopodium flabelliforme, in inhibiting the cell growth and tumor invasive behavior of DU-145 human prostate cells. In addition, the molecular action of the drug on microtubule assembly was analyzed. The effects of this diterpenoid drug in cell proliferation of DU-145 tumor cells in culture revealed that SDA at concentrations of 5 μM decreased cell growth by 14%, while at 45 μM a 61% decrease was found, as compared with control cells incubated with the solvent but in the absence of the drug. To study their effects on the cell cycle, DU-145 cells were incubated with increasing concentrations of SDA and the distribution of cell-cycle stages was analyzed by flow cytometry. Interestingly, the data showed that 14-keto-stypodiol diacetate dramatically increased the proportion of cells in the G2/M phases, and decreased the number of cells at the S phase of mitosis, as compared with appropriate controls. Studies on their action on the in vitro assembly of microtubules using purified brain tubulin, showed that SDA delayed the lag period associated to nucleation events during assembly, and decreased significantly the extent of polymerization. The studies suggest that this novel derivative from a marine natural product induces mitotic arrest of tumor cells, an effect that could be associated to alterations in the normal microtubule assembly process. On the other hand, a salient feature of this compound is that it affected protease secretion and the in vitro invasive capacity, both properties of cells from metastases. The secretion of plasminogen activator (u-PA) and the capacity of DU-145 cells to migrate through a Matrigel-coated membrane were significantly inhibited in the presence of micromolar concentrations of SDA. These results provide new keys to analyze the functional relationships between protease secretion, invasive behavior of tumor cells and the microtubule network.

Stypopodium flabelliforme 14-keto-stypodiol diacetate DU-145 cell line microtubule assembly cell proliferation plasminogen activator (u-PA) tumor invasiveness 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Bershadsky A, Vasiliev J: Factors controlling the distribution of pseudopods: Reorganization of microtubules. In: Siekevitz (ed). Cytoskeleton. Plenum Press, New York, 1988, pp 238-242Google Scholar
  2. 2.
    Maccioni RB, Cambiazo V: Role of microtubules-associated proteins in the control of microtubule assembly. Physiological Reviews 75: 835-864, 1995Google Scholar
  3. 3.
    Jordan MA, Thrower D, Wilson L: Mechanism of inhibition of cell proliferation by Vinca alkaloids. Cancer Res. 51: 2212-2222, 1991Google Scholar
  4. 4.
    Haar E, Kowalski J, Hamel E, Lin MC, Longley RE, Gunasekera S, Rosekranz H, Day B: Discodermolide a cytotoxic marine agent that stabilizes microtubules more potently than taxol. Biochemistry 35: 243-250, 1996Google Scholar
  5. 5.
    Hartley-Asp B: In: K. Harraps, W. Davis, A. Calvert (eds). Cancer Chemotherapy and Selective Drug Development. Martinus Nijhoff, Dordrecht, 1984, pp 425–429Google Scholar
  6. 6.
    Wilson L, Jordan MA: Microtubule dynamics: Taking aim at a moving target. Chem Biol 2: 569-573, 1995Google Scholar
  7. 7.
    Friden B, Rutberg M, Deinum J, Wallin M: The effect of estramustine derivatives on microtubule assembly in vitro depends on the charge of the substituent. Biochem Pharmacol 42: 997-1006, 1991Google Scholar
  8. 8.
    Moraga D, Rivas-Berrios A, Farias G, Wallin M, Maccioni RB: Estramustine-phosphate binds to a tubulin binding domain on microtubule-associated proteins Map-2 and Tau. Biochim Biophys Acta 1121: 97-103, 1993Google Scholar
  9. 9.
    Parness J, Horwitz SB: Taxol binds to polymerized tubulin in vitro. J Cell Biol 91: 479-487, 1981Google Scholar
  10. 10.
    Toso RJ, Jordan MA, Farrell KW, Matsumoto B, Wilson L: Kinetic stabilization of microtubule dynamic instability in vitro by vinblastine. Biochemistry 32: 1285-1293, 1993Google Scholar
  11. 11.
    Martínez J, Santibañez F, Vial C, Maccioni RB: The antineoplasic agent estramustine and the derivative estramustine-phosphate inhibit secretion of interleukin-3 in leukemic cells. Possible roles of MAPs. Mol Cell Biochem 117: 165-173, 1992Google Scholar
  12. 12.
    Santibañez JF, Maccioni RB, Martinez J: The secretion of urokinaselike plasminogen activator is inhibited by microtubule-interacting drugs. Cell Biochem Funct 13: 217-225, 1995Google Scholar
  13. 13.
    O'Brien E, Jacob RS, Wilson L: Inhibition of brain microtubule assembly in vitro by stypoldione. Mol Pharmacol 24: 493-499, 1983Google Scholar
  14. 14.
    O'Brien E, Asai DJ, Jacob R, Wilson L: Selective inhibition of cytokinesis in sea urchin embryos by low concentrations of stypoldione, a marine natural product that reacts with sulfhydryl groups. Mol Pharmacol 35: 1851-1855, 1986Google Scholar
  15. 15.
    Gil B, Ferrándiz ML, Sanz MJ, Terencio MC, Ubeda A, Rovirosa J, San Martin A, Alcaraz MJ, Payá M: Inhibition of inflammatory responses by epitaondiol and other marine metabolites. Life Sci 57: 25-30, 1995Google Scholar
  16. 16.
    Rovirosa J, Sepúlveda M, Quezada E, San Martin A: Isoepitaondiol, a diterpenoid of Stypopodium flabelliforme and the insecticidal activity of stypotriol, epitaondiol and derivatives. Phytochemistry 31: 2679-2681, 1992Google Scholar
  17. 17.
    Shelanski ML, Gaskin F, Cantor CR: Assembly of microtubules in the absence of added nucleotides. Proc Natl Acad Sci USA 70: 765-768, 1973Google Scholar
  18. 18.
    Vera JC, Rivas C, Maccioni RB: Biochemical dissection of the role of the l kDa C-terminal domain of tubulin subunits in the assembly of microtubules. Biochemistry 28: 333-339, 1989Google Scholar
  19. 19.
    Gaskin F, Cantor CR, Shelanski NC: Turbidimetric studies of the in vitro assembly and disassembly of microtubules. J Mol Biol 89: 736-773, 1974Google Scholar
  20. 20.
    Stone KR, Mickey D, Wunderly H, Mickey GH, Paulson DF: Isolation of a human prostate carcinoma cell line DU-145. Int J Cancer 21: 274-281Google Scholar
  21. 21.
    Cross D, Vial C, Maccioni RB: A tau-like protein interacts with stress fibers and microtubules in human and rodent cell lines. J Cell Sci 105: 51-60, 1993Google Scholar
  22. 22.
    Crissman HA, Steinkamp JA: Rapid, simultaneous measurement of DNA, protein and cell volume in single cells from large mammalian populations. J Cell Biol 59: 776-771, 1973Google Scholar
  23. 23.
    Carmichel J, De Graff WG, Gazdar AF, Minna JD, Mitchell JB: Evaluation of a tetrazolium-based semiautomated colorimetric assay: Assessment of chemosensitivity testing. Cancer Res 47: 936-942, 1987Google Scholar
  24. 24.
    Gerwick WH, Fenical W: Ichtyotoxic and cytotoxic metabolism of the tropical brow algae Stypopodium zonale Papenfus. J Org Chem 46: 22-27, 1981Google Scholar
  25. 25.
    White SJ, Jacob RS: Effect of stypoldione on cell cycle progression, DNA and protein synthesis, and cell division in cultured sea urchin embryos. Mol Pharmacol 24: 500-508, 1983Google Scholar
  26. 26.
    Jordan MA, Thrower D, Wilson L: Effects of vinblastine, phodophyllotoxin and nocodazole on mitotic spindles. J Cell Sci 102: 401-416, 192Google Scholar
  27. 27.
    González M, Cambiazo V, Maccioni RB: Identification of a new microtubule-interacting protein Mip-90. Eur J Cell Biol 67: 158-169, 1995Google Scholar
  28. 28.
    Henriquez JP, Cross D, Vial C, Maccioni RB: Subpopulations of tau interact with microtubules and actin filaments in various cell types. Cell Biochem Funct 13: 239-250, 1995Google Scholar
  29. 29.
    Schpetner HS, Paschal BM, Vallee RB: Characterization of the microtubule-activated ATPase of brain cytoplasmic dynein (MAP-1C). J Cell Biol 197: 1001-1009, 1988Google Scholar
  30. 30.
    Jacob RS, Culver P, Langdon R, O'Brien T, White S: Some pharmacological observations on marine natural products. Tetrahedron 41: 981-984, 1985Google Scholar
  31. 31.
    McNally FJ: Modulation of microtubule dynamics during the cell cycle. Curr Opinion Cell Biol 8: 23-29, 1996Google Scholar

Copyright information

© Kluwer Academic Publishers 1998

Authors and Affiliations

  • María S. Depix
    • 1
    • 2
  • Jorge Martínez
    • 3
  • Francisco Santibañez
    • 3
  • Juana Rovirosa
    • 4
  • Aurelio San Martín
    • 4
  • Ricardo B. Maccioni
    • 1
    • 2
  1. 1.Laboratory of Cellular and Molecular BiologyUniversity of Chile, Las Palmeras, ÑuñoaSantiagoChile
  2. 2.International Center for Cancer and Developmental Biology (ICC), Las Palmeras, ÑuñoaSantiagoChile
  3. 3.Institute of Nutrition and Food Technology (INTA)University of Chile, Las Palmeras, ÑuñoaSantiagoChile
  4. 4.Department of Chemistry, Faculty of SciencesUniversity of Chile, Las Palmeras, ÑuñoaSantiagoChile

Personalised recommendations