The Cytolytic Effect of a Glycoconjugate Extracted from Corms of Saffron Plant (Crocus sativus) on Human Cell Lines in Culture

Abstract

Corms of Crocus sativus L. (Iridaceae) contain a glycoconjugate that shows cytotoxic activity on tumoral cells in culture. Studies of intracellular calcium fluctuations, and release of lactate dehydrogenase in human cervical epitheloid carcinoma cells, showed that this compound caused plasma membrane damage, allowing movements of both calcium and macromolecules, and leading to cell lysis. Analysis of DNA fragmentation showed that cell death was not mediated by apoptosis. This molecule is active against human tumoral cells derived from fibrosarcoma, cervical epithelioid carcinoma and breast carcinoma, with IC₅₀ values of 7, 9 and 22 μg/ml, respectively. The proteoglycan is about 8 times more cytotoxic for malignant cells than for their normal counterparts. In addition, 100 μg/ml of proteoglycan produced 50 % in vitro lysis of normal human erythrocytes, whereas 320 μg/ml induced about 60 % cell death on cultured human hair follicles. Altogether, these results suggests a distinctive cytotoxic activity of this molecule on different human cell types.

Key words

Crocus sativus - Iridaceae - cytolysis - cytotoxicity - proteoglycan - glycoconjugate

References

• 1 Mueller  E A,, Anderer  F A.. Chemical specificity of effector cell/tumor cell bringing by a Viscum album rhamnogalacturonan enhancing cytotoxicity of human NK cells.  Immunopharmacol.. 1990;;  19 69-77
• 2 Mueller  E A,, Anderer  F A.. Synergistic action of a plant rhamnogalacturonan enhancing antitumor cytotoxicity of human natural killer cells: chemical specificity of target cell recognition.  Cancer Res.. 1990;;  50 3646-51
• 3 Hauer  J,, Anderer  F A.. Mechanism of stimulation of human natural killer cytotoxicity by arabinogalactan from Larix occidentalis.  Cancer immunol. Immunother.. 1993;;  36 237-224
• 4 Yamada  H,, Komiyama  K,, Kiyohara  H,, Cyong  J-C,, Hirakawa  Y,, Otsuka  Y.. Structural characterization and antitumor activity of a pectic polysaccharide from the roots of Angelica acutiloba.  Planta Med.. 1990;;  56 182-6
• 5 Hagmar  B,, Ryd  W,, Skomedal  H.. Arabinogalactan blockade of experimental metastases to liver by murine hepatoma.  Invas. Metast.. 1991;;  11 348-55
• 6 Leibovici  J,, Stark  Y.. Slow cytotoxicity of the polysaccharide levan on tumor cells in vitro.  Chem. Biol. Interact.. 1986;;  60 191-200
• 7 Escribano  J,, Ríos  I,, Fernández  J A.. Isolation and cytotoxic properties of a novel glycoconjugate from corms of saffron plant (Crocus sativus L.).  Biochim. Biophys. Acta. 1999;;  1426 217-22
• 8 Haugland  R P.. Handbook of fluorescent probes and research chemicals. Molecular Probes Europe BV, Leiden, The Netherlands; 1996
• 9 Martinez-Zaguilan  R,, Parnami  G,, Lynch  R M.. Selection of fluorescent ion indicator for simultaneous measurements of pH and Ca²⁺.  Cell calcium. 1996;;  19 337-49
• 10 Videlov  L L,, Christensen  I J,, Nissen  N I.. A detergent-trypsin method for the preparation of nuclei for flow cytometry DNA analysis.  Cytometry. 1983;;  3 323-27
• 11 Bergmeyer  H U,, Grabl  M,, Walter  H-E.. Methods of enzymatic analysis,. (Bergmeyer HU, eds.) 232-233, Weinheim Verlag Chemie, Germany; 1984
• 12 Velasco  M,, Díaz-Guerra  M JM,, Díaz-Achirica  P,, Andreu  D,, Rivas  L,, Bosch  L.. Macrophage triggering with cecropin A and melittin-derived peptides induces type II nitric oxide synthase expression.  J. Immunol.. 1997;;  158 4437-43
• 13 Raynor  R L,, Zeng  B Z,, Kuo  J F.. Membrane interactions of amphiphilic polypeptides mastoparan, melittin, polymyxin B, and cardiotoxin. Differential inhibition of protein kinase C, Ca²⁺/calmodulin-dependent protein kinase II and synaptosomal membrane Na,K-ATPase, and Na⁺ pump and differentiation of HL60 cells.  J. Biol. Chem.. 1991;;  266 2753-58
• 14 Gupta  A,, Hruska  K A,, Wedner  H J.. Phytohemagglutinin rapidly lyses S49 T-lymphoma cells and the cytotoxicity is not mediated by generation of cAMP or increase in cytosolic calcium.  Biochem. Biophys. Res. Commun.. 1991;;  170 1035-43
• 15 Saberwal  G,, Nagaraj  R.. Cell-lytic and antibacterial peptides that act by perturbing the barrier function of membranes: facets of their conformation features, structure-function correlation and membrane-perturbing abilities.  Biochim. Biophys. Acta. 1994;;  1197 109-31
• 16 Liebmann  J E,, Cook  J A,, Lipschultz  C,, Teague  D,, Fisher  J,, Mitchell  J B.. Cytotoxic studies of paclitaxel (Taxol) in human tumor lines.  Br. J. Cancer. 1993;;  68 1104-09
• 17 Kohn  E C,, Felder  C C,, Jacobs  W,, Holmes  K A,, Day  A,, Freer  R,, Liotta  L A.. Structure-function analysis of signal and growth inhibition by carboxyamidotriazole, CAI.  Cancer Res.. 1994;;  54 935-42
• 18 Stein  C A.. Suramin, a novel antineoplastic agent with multiple potential mechanisms of action.  Cancer Res.. 1993;;  53 2239-48