Abstract
In recent years, sulfated fucans have emerged as an important class of natural biopolymers.
In this study, the anti-adhesive activity of a fucan from the brown seaweed Spatoglossum schröederi was analyzed using tumorigenic cells: wild-type Chinese hamster ovary cells (CHO-K1)
and the mutant type deficient in xylosyltransferase (CHO-745). Fibronectin (FN) was
used as substrate for cell attachment. For both cell types, this fucan has shown a
dose-dependent anti-adhesive effect, reaching saturation at around 400 μg/mL. This
effect was abolished by desulfation of the fucan. In addition, this polymer exhibited
the highest inhibitory effect in comparison to other sulfated polysaccharides. The
fucan was biotinylated and used as a probe to identify its action sites. Biotinylated
fucan was detected in the extracellular matrix environment by confocal microscopy
and flow cytometric analysis, but not at the cell surface. The results suggest that
the fucan shows anti-adhesive activity by binding directly to FN, and blocking FN
sites that are recognized by cell surface ligands, possibly the integrin family.
Key words
CHO cells - cell adhesion - anti-adhesive fucan - fucoidan -
Spatoglossum schröederi
- Phaeophyceae
References
1
Dietrich C P, Farias G GM, Abreu L RD, Leite E L, Silva L F, Nader H B.
A new approach for characterization of polysaccharides from algae: Presence of four
main acidic polysaccharides in three species of the class Phaeophycea.
Plant Sci.
1995;
108
143-53
2
Vilela-Silva E SA, Castro M O, Valente A P, Biermann H C, Mourão P AS.
Sulfated fucans from the egg jellies of the closely related sea urchins Strongylocentrotus droebachiensis and Strongylocentrotus pallidus ensure species-specific fertilization.
J Biol Chem.
2002;
277
379-87
3
Soeda S, Ohmagugi Y, Shimeno H, Nagamatsu A.
Preparation of aminated fucoidan and its evaluation as an antithrombotic and antilipemic
agent.
Biol Pharmac Bull.
1994;
17
784-8
4
Liu J M, Haroun-Bouhedja F, Boisson-Vidal C.
Analysis of the in vitro inhibition of mammary adenocarcinoma cell adhesion by sulfated polysaccharides.
Anticancer Res.
2000;
20
3265-72
5
Leite E L, Medeiros M GL, Rocha H AO, Farias G GM, Silva L F, Chavante S F. et al
.
Structure and pharmacological activities of sulfated xylofucoglucuronan from the alga
Spatoglossum schröederi
.
Plant Sci.
1998;
132
215-28
6
Rocha H AO, Franco C RC, Trindade E S, Carvalho L CM, Veiga S S, Leite E L. et al
.
A fucan from the brown seaweed Spatoglossum schröederi inhibits Chinese hamster ovary cell adhesion to several extracellular matrix proteins.
Braz J Med Biol Res.
2001;
34
621-6
7
Dubois M, Gilles K A, Hamilton J K, Rebers P A, Smith F.
Colorimetric method for determination of sugars and related substances.
Anal Chem.
1956;
28
250-6
8
Nader H B, Dietrich C P.
Determination of sulfate after chromatography and toluidine blue complexation.
Anal Biochem.
1977;
78
112-8
9
Spector J.
Refinement of the coomassie blue method of protein quantification. A simple and linear
spectrophotometric assay of 0.5 to 50 μg of protein.
Anal Biochem.
1978;
86
142-3
10
Kozakai M, Yosizawa Z.
A rapid method for separation and identification of several hexuronic acids and hexuronic
acid-containing oligosaccharides.
Anal Biochem.
1975;
78
425-9
11
Trindade E S, Oliver C, Jamur M C, Franco C RC, Gouveia T C, Dietrich C P, Nader H B.
Biotinylated heparin stimulates heparan sulfate production without binding the cell
surface.
Mol Biol Cell.
1999;
10 (n. S.)
459
12
Rapraeger A C, Krufka A, Bradley B O.
Requirement of heparan sulfate for βFGF-mediated fibroblast growth and myoblast differentiation.
Science.
1991;
252
1705-7
13
Raimondi L, Banchelli G, Dalmazzi D, Mulinacci N, Romani A, Vincieri F F. et al .
Sedum telephium L. polysaccharide content affects MRC5 cell adhesion to laminin and fibronectin.
J Pharm Pharmacol.
2000;
20
585-91
14
Esko J D, Rostand K S, Weinke J L.
Tumor formation dependent on proteoglycan biosynthesis.
Science.
1988;
241
1092-6
15
Franco C RC, Rocha H AO, Trindade E S, Santos I AN, Leite E L, Veiga S S. et al .
Heparan sulfate and control of cell division: adhesion and proliferation of mutant
CHO-745 cells lacking xylosyl transferase.
Braz J Med Biol Res.
2001;
34
971-5
Helena B. Nader
Departamento de Bioquímica
Escola Paulista de Medicina
Universidade Federal de São Paulo
Rua 3 de Maio 100
CEP 04044-020
São Paulo SP
Brazil
Phone: +55-11-5579-3175
Fax: +55-11-5573-6407
Email: hbnader.bioq@epm.br