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Anti-adhesive fucan
ISSN 0100-879X
C.P. Dietrich3 and Universidade Federal de São Paulo, São Paulo, SP, Brasil
H.B. Nader3
Abstract
Correspondence Fucans, a family of sulfated polysaccharides present in brown sea- Key words
H.B. Nader weed, have several biological activities. Their use as drugs would · CHO cells
Departamento de Bioquímica offer the advantage of no potential risk of contamination with viruses · Extracellular matrix proteins
EPM, UNIFESP
or particles such as prions. A fucan prepared from Spatoglossum · Fucan
Rua 3 de Maio, 100, 4º andar Glycosaminoglycans
04044-020 São Paulo, SP
schröederi was tested as a possible inhibitor of cell-matrix interac- ·
· Cell adhesion
Brasil tions using wild-type Chinese hamster ovary cells (CHO-K1) and the
· Brown algae
Fax: +55-11-573-6407 mutant type deficient in xylosyltransferase (CHO-745). The effect of
E-mail: hbnader.bioq@epm.br this polymer on adhesion properties with specific extracellular matrix
components was studied using several matrix proteins as substrates for
Presented at cell attachment. Treatment with the polymer inhibited the adhesion of
SIMEC 2000 - International
fibronectin to both CHO-K1 (2 x 105) and CHO-745 (2 x 105 and 5 x
Symposium on Extracellular
105) cells. No effect was detected with laminin, using the two cell
Matrix, Angra dos Reis, RJ,
Brazil, September 24-27, 2000.
types. On the other hand, adhesion to vitronectin was inhibited in
CHO-K1 cells and adhesion to type I collagen was inhibited in CHO-
Research supported by FAPESP, 745 cells. In spite of this inhibition, the fucan did not affect either cell
CAPES and CNPq. proliferation or cell cycle. These results demonstrate that this polymer
is a new anti-adhesive compound with potential pharmacological
applications.
Received September 22, 2000
Accepted March 6, 2001
Introduction tested for biological activities in different
mammalian systems. Alga fucans have anti-
Fucan is a term used to define a family of viral (3), anti-ulcer and anti-adhesion activ-
L-fucose-containing sulfated polysaccharides ity (4), anticoagulant activity (1,5), anti-in-
with extremely variable molecular weights. flammatory activity, and antiproliferative and
They are extracted from the extracellular antitumoral properties. They are inhibitors
matrix of brown seaweed (1), the jelly coat of vascular smooth muscle cell and fibro-
of sea urchin eggs (1), and the body wall of blast proliferation and can block sperm-egg
the sea cucumber (2). binding in several species (6). The structures
Since the first description of the fucan of these fucans vary among species and some-
from algae, these polysaccharides have been times among different parts of the plant (7).
On the other hand, in contrast to animal the average structure of a 56-kDa fucan.
fucans, alga fucans have portions of other Although chemical hydrolysis is important
neutral sugars and uronic acids in addition to to determine the structure of fucans, it can
sulfate and fucose in their structures. This cause structural alterations such as debranch-
complexity led Grauffel et al. (5) to propose ing or desulfation. Only enzymatic methods
that the chemical composition of the differ- can cleave glycosidic linkages specifically
ent polysaccharides may vary according to without modifying the structural units pre-
the method of extraction. Thus, each new sent in the original polysaccharide. How-
fucan purified is a unique compound with ever, no commercial endofucosidases are
unique structural features and thus a poten- available. The extraction of LMW fucans
tial new drug. from brown seaweed, which have biological
activity, would be very important to deter-
Structural features of fucans mine the structure/biological activity rela-
tionships of fucans.
The structural characteristics of sulfated Dietrich et al. (7) have fractionated fucans
fucans responsible for all these biological from three types of brown seaweeds using
activities have not been determined and con- the same methodology for the characteriza-
sequently the relationships between struc- tion and quantitation of the different classes
tural and biological activities have not been of glycosaminoglycans (11). They have dem-
clearly established. Most of the difficulties onstrated that these seaweeds contain three
for these studies arise from the fact that these main fucans with distinct electrophoretic mo-
compounds are highly heterogeneous poly- bilities which were named A, B and C ac-
saccharides which give complex nuclear mag- cording to their relative mobility. A 21-kDa
netic resonance (NMR) spectra with broad LMW fucan with mobility equal to that of
signals hampering resolution (1). In fact, for fucan A was extracted from the brown algae
these plant polysaccharides even high-field Spatoglossum schröederi (10) using the same
NMR is of limited value, and complete de- methodology. This fucan is composed of a
scriptions of their structures are not avail- core of ß (1-3) glucuronic acid-containing
able at present (2). However, meaningful oligosaccharide of 4.5 kDa with branches at
structural studies using NMR are possible C-4 of fucose chains a (1-3) linked. The
with relative low molecular weight (LMW) fucose is mostly substituted at C-4 with sul-
fucans that have been prepared by several fate groups and at C-2 with chains of ß (1-4)
methods such as acid hydrolysis of high xylose, which, in turn, is also partially sul-
molecular weight (HMW) fucans (8,9) or fated. Recently, using this methodology we
extration from brown seaweeds (10). have also purified a fucan with the mobility
Studies with LMW fucans obtained by of fucan B from the same alga (12). This
chemical hydrolysis of HMW fucans have compound differs from the A fucan by the
demonstrated that 3- and/or 4-linked fucose presence of significant amounts of galactose.
residues are always present, sulfation at the Here we describe the effect of this fucan
2 and 4 positions can also be identified in on cell proliferation, cell cycle and adhe-
these polymers and their biological activities siveness of both wild-type Chinese hamster
are apparently related not only to molecular ovary cells (CHO-K1) and xylosyltrans-
weight and sulfation, but also the quantity ferase-deficient mutant (CHO-745) to sev-
and position of the sulfated residues in the eral extracellular matrix proteins. In the mu-
compound (1,6,8). Recently, Nagaoka et al. tant the synthesis of both heparan and chon-
(4), using data from partial acid hydrolysis, droitin sulfates is diminished. The glycosami-
methylation and NMR analysis, proposed noglycans synthesized by mutant 745 were
about 5% of those produced by wild-type distribution during the phases of the cell
cells (13). CHO-K1 cells are tumorigenic cycle (Figure 1A) when quiescent cultures
whereas CHO-745 mutant cells are not (13). of both cell lines were incubated with [3H]-
thymidine (0.25 µCi/ml), 10% FCS and fucan
Cell proliferation and fucans
A
Cell proliferation is a physiological pro- 80000
cess regulated by a broad range of growth K1 + FCS
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