Cytotoxic Activity of the Seeds from Agrostemma githago var. githago

Abstract

In our investigations aqueous extracts of the seeds from Agrostemma githago L. proved to be remarkably more cytotoxic in comparison with isolated agrostemmasaponins in equal concentrations. A combination of agrostemmasaponin 1 with a formyl function attached to triterpene position 4 together with agrostin, a ribosome-inactivating protein (RIP type 1; M_r: 27 kDa) which we isolated from the seed material, resulted in a markedly increased cytotoxicity. In analogy to the well-known lectin-like mechanism of action which can be inhibited by different monosaccharides, we tested seven different sugars known for their affinity to prevent cell recognition and thereby detoxify lectins. None of the tested monosaccharides diminished toxicity values significantly. This suggests that agrostin, combined with agrostemmasaponin 1, has a different mode of action to penetrate through the cell membrane than lectins. On the basis of these facts we concluded that both compounds work as a functional unity showing a mechanism of ”cooperative toxicity”.

Agrostemma githago L. - Caryophyllaceae - seeds - agrostemmasaponin 1 - agrostin - ribosome-inactivating protein - membrane - cytotoxicity

References

• 1 Kruskal N. Ueber Agrostemma githago L.  Dissertation. Universität Dorpat 1891
• 2 Melzig M F, Bader G, Loose R. Investigations of the mechanism of membrane activity of selected triterpenoid saponins.  Planta Med. 2001;  67 43-8
• 3 Siepmann C, Bader G, Hiller K, Wray V, Domke T, Nimtz M. New saponins from the seeds of Agrostemma githago L.  Planta Med. 1998;  64 159-64
• 4 Stirpe F, Gasperi-Campani A, Barbieri L, Falasca A, Abbondanza A, Stevens W A. Ribosome-inactivating proteins from the seeds of Saponaria officinalis, Agrostemma githago, Asparagus officinalis and from the latex of Hura crepitans .  Biochem J. 1983;  216 617-25
• 5 De Klerk G J, Engelen D. Assembly of Agrostemma githago storage proteins and their precursor proteins into oligomers.  Biochem J. 1985;  229 269-72
• 6 Hiegemann M, Duddeck H, Bader G, Hiller K. Structural investigation and conformational analysis of helianthosid 2, a saponin from Helianthus annuus .  Magn Reson Chem. 1995;  33 337-41
• 7 Bradford M M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.  Anal Biochem. 1976;  72 248-54
• 8 Smith P K, Krohn R I, Hermanso G T, Mallia A K, Gartner F H, Provenzano M D, Fujimoto E K, Goeke N M. et al . Measurement of protein using bicinchoninic acid.  Anal Biochem. 1985;  150 76-85
• 9 Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays.  J Immunol Methods. 1983;  65 55-63
• 10 Hoffmann-Bohm K. Hagers Handbuch der Pharmazeutischen Praxis. Folgeband 4, 5th Edition Springer-Verlag Berlin, Heidelberg; 1992: pp. 142-5
• 11 Barr I G, Sjölander A, Cox J C. ISCOMS and other saponin-based adjuvants.  Adv Drug Deliv Rev. 1998;  32 247-71
• 12 Bomford R, Stapleton M, Windsor S, Beesley J E, Jessup E A, Price K R, Fenwick G R. Adjuvanticity and ISCOM formation by structurally diverse saponins.  Vaccine. 1992;  10 572-7
• 13 Pillion D J, Amsden J A, Kensil C, Recchia J. Structure-function relationship among quillaja saponins serving as excipients for nasal and ocular delivery of insulin.  J Pharm Sci. 2000;  85 518-24
• 14 Chao A C, Nguyen J V, Broughall M, Recchia J, Kensil C R, Daddona P E, Fix J A. Enhancement of intestinal model compound transport by DS-1, a modified Quillaja saponin.  J Pharm Sci. 1998;  87 1395-9
• 15 Soltysik S, Wu J Y, Recchia J, Wheeler D A, Newman M J, Coughlin R T, Kensil C. Structure/function studies of QS-21 adjuvant: assessment of triterpene aldehyde and glucuronic acid roles in adjuvant function.  Vaccine. 1995;  13 1403-10
• 16 Kamstrup S, San Martin R, Doberti A, Grande H, Dalsgaard K. Preparation and characterisation of Quillaja saponin with less heterogeneity than Quil-A.  Vaccine. 2000;  18 2244-9
• 17 Gaidi G, Correia M, Chauffert B, Beltramo J, Wagner H, Lacaille-Dubois M A. Saponins-mediated potentiation of cisplatin accumulation and cytotoxicity in human colon cancer cells.  Planta Med. 2002;  68 70-2
• 18 Barbieri L, Battelli M G, Stirpe F. Ribosome-inactivating proteins from plants.  Biochim Biophys Acta. 1993;  1154 237-82
• 19 Rhodes J. Evidence for an intercellular covalent reaction essential in antigen-specific T cell activation.  J Immunol. 1989;  143 1482-9