Journal of Dental Research

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ABO Blood-group Antigens in Oral Cancer

E. Dabelsteen and S. Gao
J DENT RES 2005 84: 21
DOI: 10.1177/154405910508400103

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 CRITICAL REVIEWS IN ORAL BIOLOGY & MEDICINE
ABO Blood-group Antigens in Oral Cancer
E. Dabelsteen* and S. Gao
Department of Oral Diagnostics, School of Dentistry,
University of Copenhagen, Nørre Alle 20, DK-2200
Copenhagen N, Denmark; *corresponding author,
ed@odont.ku.dk
J Dent Res 84(1):21-28, 2004
ABSTRACT
Tumor progression is often associated with altered
glycosylation of the cell-surface proteins and
lipids. The peripheral part of these cell-surface
glycoconjugates often carries carbohydrate
structures related to the ABO and Lewis blood-
group antigens. The expression of histo-blood-
group antigens in normal human tissues is
dependent on the type of differentiation of the
epithelium. In most human carcinomas, including
oral carcinoma, a significant event is decreased
expression of histo-blood-group antigens A and B.
The mechanisms of aberrant expression of blood-
group antigens are not clear in all cases. A relative
down-regulation of the glycosyltransferase that is
involved in the biosynthesis of A and B antigens is
seen in oral carcinomas in association with tumor
development. The events leading to loss of A
transferase activity are related, in some instances,
to loss of heterozygosity (LOH) involving
chromosome 9q34, which is the locus for the ABO
gene, and in other cases, to a hypermethylation of
the ABO gene promoter. The fact that hyper-
methylation targets the ABO locus, but not
surrounding genes, suggests that the hyper-
methylation is a specific tumor-related event.
However, since not all situations with lack of
expression of A/B antigens can be explained by
LOH or hypermethylation, other regulatory factors
outside the ABO promoter may be functional in
transcriptional regulation of the ABO gene.
Altered blood group antigens in malignant oral
tissues may indicate increased cell migration. This
hypothesis is supported by studies showing that
normal migrating oral epithelial cells like
malignant cells show lack of expression of A/B
antigens, and by studies that target ABH antigens
to key receptors controlling adhesion and motility,
such as integrins, cadherins, and CD-44.
KEY WORDS: oral cancer, ABO blood-group
antigens, carbohydrate antigens, oral precancer.
Received January 27, 2004; Accepted June 7, 2004
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INTRODUCTION
Tto heboundary
membrane, which defines the extent of the cell, is not only a physical
but also has many specific functions, among which is the capacity
react with other cells and the intracellular matrix (Ebnet and Vestweber,
1999; Hascall, 2000). Carbohydrates are structures found on the cell surface
bound to either lipid or protein embedded in the membrane. Changes in the
carbohydrate structure of these cell-surface glycolipids and glycoproteins have
been demonstrated during development, during cell maturation in adult tissue,
and in relationship to malignant development (Fenderson et al., 1986;
Dabelsteen, 1996; Hakomori, 1999, 2002, 2003; Le Pendu et al., 2001;
Fukuda, 2002). The cell-surface carbohydrates have an enormous potential for
serving as informational molecules. Monosaccharides can be linked together in
different sequences and by different glycosidic linkages, thereby generating a
vast complexity of saccharide chains. The variety of structures, which can be
formed by a limited number of units, is thus far larger in oligosaccharide chains
than in peptides. Each monosaccharide unit has 3 or 4 different sites that can be
substituted by the next sugar, in contrast to amino acids, which have only one
site for linkage with the next monomeric unit (Roseman, 2001). Although
carbohydrates are extremely complex structures, the cellular expression is
highly regulated, and, within a particular organ, the carbohydrates may be
expressed in a way that correlates with cell differentiation. When carbo-
hydrates are bound to proteins, changes in glycosylation may affect the
conformation and function of the protein and thereby influence the interaction
between the cell and its environment. It has been demonstrated, for example,
that changes in glycosylation of integrins alter their function, and that the
Notch receptor, which is a transmembrane protein that mediates
communication associated with cell differentiation, is regulated by alteration in
the glycosylation of the protein (Prokopishyn et al., 1999; Bruckner et al.,
2000; Moloney et al., 2000).
Cell-surface carbohydrates are built up in a stepwise fashion when
monosaccharides are tranferred from their sugar nucleotide derivatives to
appropriate acceptors. Each particular type of transfer is catalyzed by a unique
specific glycosyltransferase. Thus, a missing or changed enzyme will block
further synthesis of the carbohydrate structure, which will remain as a
precursor structure. Additionally, the appearance of new enzymes that either
compete for common substrate or lead to elongation of previously terminated
structures will result in alteration in the carbohydrate that is finally expressed
(Fukuda et al., 1999; Fukuda, 2002; Fuster et al., 2003).
In tumors, changes in glycosylation are found in both glycolipids and
glycoproteins (Hakomori, 1999; Le Pendu et al., 2001). Most studies have
dealt with alteration of carbohydrates at the cell surface. However, several
recent studies have shown that altered glycosylation plays a major role in
most aspects of the malignant phenotype, including signal transduction and
apoptosis. These studies have recently been reviewed (Hakomori, 2002;
Hakomori and Handa, 2002).
ABO BLOOD-GROUP ANTIGENS
Structure and Genetics
The peripheral part of both cell-surface glycoproteins and glycolipids often
carries carbohydrate structures related to the ABO and Lewis blood-group
antigens (Hakomori et al., 1967). Such antigens were originally ascribed to
21
22 Dabelsteen & Gao
Figure 1. The ABH determinants are composed of L-fucose, D-
galactose, and D-N-acetylgalactosamine, with a minimal determinant
structure and the synthetic pathway as described in the text (Yamamoto
et al., 1990).
erythrocytes, but have later been found on other predominantly
epithelial and endothelial cells (Hartmann, 1941; Szulman,
1960). Many tumor-associated changes are related to these
antigens and their structural precursors and were described as
early as 1930 by Thomsen and co-workers and later, were
extensively studied and reviewed by Hakomori (Hakomori,
1985, 1999, 2002).
Two carbohydrate antigens, A and B antigens, and their
antibodies constitute the ABO system (Watkins, 1966). In
recent years, the molecular basis for the ABO system has been
described in detail (Yamamoto et al., 1990; Clausen et al.,
1994; Yamamoto, 2000). The A and B functional alleles of the
ABO genetic locus encode the A transferase (glycosyl-
transferase ␣1,3-GalNAc-transferase) and the B transferase
(␣1,3Gal-transferase), respectively. The A transferase transfers
a GalNAc residue from UDP-GalNAc to the precursor H
substrate, producing A antigens as defined by the trisaccharide
determinant structure G␣lNAc-␣1,3 (Fuc␣1-2)GalNAc␤1-R
(Fig. 1). Similarly, the B transferase catalyzes the transfer of
Gal from UDPGal to the H substrate, producing B antigens
(Fig. 1). The phenotypic differences between A and B are thus
due to the subtle differences in substrate specificity of the A
and B glycosyltransferases. Phenotype O is characterized by
the absence of A and B transferase and, most likely, by the
presence of an inert protein encoded by the O allele, leaving the
H structure unchanged. The synthesis of the H structure is
controlled by 2 fusosyltransferases coded for by 2 distinct
genes, Fut1 and Fut2 (Koda et al., 1997a,b; Oriol et al., 2000).
Fut1 controls the expression of H on erythrocytes, endo-
thelium, and epithelial cell membranes, whereas Fut2 is
responsible for H in saliva and other secretions, but also on
some epithelial cell membranes (Oriol et al., 2000). Secretors
are defined as individuals that secrete blood-group antigen H or
A/B in saliva. The Fut2 gene is absent in 20% of the Western
population, resulting in the absence of ABH antigens in saliva.
Secretors were classically designated as SeSe or Sese, and non-
secretors as sese (Hartmann, 1941; Watkins, 1966; Rouquier et
al., 1995; Oriol et al., 2000).
The molecular basis of the histo-blood-group antigens was
elucidated by isolation of the A transferase and cloning of the
cDNA encoding this enzyme (Clausen et al., 1990; Yamamoto
et al., 1990). Three nucleotide substitutions defined the
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J Dent Res 84(1) 2005
common difference between A and B allelic cDNA. Several
blood-group A subtypes have been identified, and the A
transferase isolated from blood-group A 1 subtype is
qualitatively different from the A transferase isolated from
subtype A2. This is reflected in the nucleotide sequence of the
A gene and illustrates the polymorphic nature of the ABO genes
(Yamamoto et al., 1990; Hakomori, 1999; Olsson et al., 2001).
The most common O allelic cDNA sequence is identical to the
A allele, except for the single base dilution, which indicates
that lack of transferase activity in O individuals is due to a shift
in the reading frame, leading to a functional failure (Yamamoto
et al., 1990). However, other variants of O allele exist,
including rare variants with substitutions rather than deletions
(Clausen et al., 1994). The genetic difference between A/B and
O can be detected by restriction enzymes (Fig. 2) (Yamamoto
et al., 1990). Genetic analyses by such restriction enzymes
have been used to detect cancer-associated genetic changes in
carcinoma cells (Yamamoto et al., 1990; Orlow et al., 1998).
Tissue Localization
Most studies concerning the tissue localization of the histo-
blood-group antigens have shown that the antigens in the
tissues correspond to the erythrocyte blood group, but that the
tissue expression is dependent on the secretor status of the
individual (Ravn and Dabelsteen, 2000). Furthermore, the
expression of histo-blood-group antigens in normal human
tissues is dependent on the type of differentiation of the
epithelium, i.e., simple epithelia vs. stratified epithelia, and the
degree of maturation of the single cell within the epithelium. In
stratified epithelium, the expression of histo-blood-group
antigens depends on the state of cellular differentiation
(maturation), and there is a sequential elongation of the
terminal carbohydrate chain during the life span of the cell.
Basal cells express short carbohydrate chains that are A/B
precursors, whereas A or B antigens may be seen in the spinous
cell layer. Variation in differentiation pattern—for example,
keratinized vs. non-keratinized—influences the expression of
blood-group antigens. Keratinized squamous epithelium may
express A or B antigens in only a very few and highly
differentiated cells, leaving the precursor H antigen expressed
on most spinous cells. In contrast, in non-keratinized
epithelium of the buccal mucosa, the precursor H is expressed
only on a few parabasal cells, whereas expression of A and B
antigens is seen in most spinous cells (Vedtofte et al., 1984;
Ravn and Dabelsteen, 2000). Immunohistochemical staining of
A and B glycosyltransferases shows that there is
correspondence between expression of A and B antigens and A
and B glycosyltransferases. Only in non-secretors do buccal
epithelial cells express A/B transferases, but no A/B products,
due to the lack of the precursor H antigen. The expression of
A/B antigens in oral tissues is thus regulated by the expression
of the A/B transferases and the availability of a substrate for the
transferase (Mandel et al., 1992).
Cancer and Precancer
Reduction or complete loss of antigen expression has been
reported in carcinomas of the oral cavity, lung, stomach, colon,
larynx, ovary, prostate, bladder, and breast (Le Pendu et al.,
2001). Loss of blood-group antigen expression may also be
seen in potentially malignant lesions of the gastric mucosa,
prostate, cervical mucosa, epithelial hyperplasia of the breast,
and oral mucosa (Le Pendu et al., 2001). Loss of A and B
J Dent Res 84(1) 2005 ABO Blood-group Antigens in Oral Cancer 23

antigens is often correlated with

an increased H antigen expression
in endometria, prostate, breast,
and oral carcinomas, whereas it is
accompanied by an increase in Ley
expression in lung and ovary
carcinomas. The presence of H
and Le y in the tumors indicates
that loss of A or B antigens is the
result of a blocked synthesis (Fig.
2) (Dabelsteen et al., 1983; Bryne
et al., 1991; Le Pendu et al.,
2001).
Immunohistochemical studies
of oral squamous cell carcinomas
have shown loss of expression of A
or B antigens in more than 80% of
cases, all of which showed
concomitant loss of A/B
transferase (Figs. 3A, 3B, 3C) (Gao
et al., 2004a,b). Studies of
potentially malignant lesions have
shown loss of A/B antigen in most
lesions with epithelial dysplasia
and in half of the lesions clinically Figure 2. Genotyping by diagnostic restriction enzyme digestion. The single-based deletion associated
classified as leukoplakias but with O alleles (position 258) creates a Kpnl site (O allele) and eliminates the BstEII site (A/B allele). Three
or 4 nucleotide substitutions between A and B allelic cDNA can also be identified by diagnostic enzymes.
without histological evidence of For genotyping, polymerase chain-reaction (PCR)-amplified DNA was analyzed by diagnostic enzyme
epithelial dysplasia (Dabelsteen et digestion (Yamamoto et al., 1990).
al., 1975; Gao et al., 2004a). As
mentioned above, in the normal
oral cavity, keratinized epithelium
in the palate or gingiva shows little or no expression of A or B restriction enzyme analysis, it was possible to genotype tumor
blood-group antigen (Dabelsteen et al., 1991). Since a change tissue and to detect specific allelic loss in oral carcinomas
from a non-keratinized to a keratinized differentiation pattern is a (Figs. 3E, 3F, 3G). From an investigation of formalin-fixed and
characteristic of many oral carcinomas and potentially malignant paraffin-embedded materials, it appeared that 1/3 of the
lesions, the lack of expression of blood-group antigens in such carcinomas showed specific loss of A and B alleles; whereas in
lesions could be due to a change in differentiation pattern of the another study, based on frozen sections only, 1/6 of the cases
epithelium (Dabelsteen et al., 1975). However, it has been had specific allelic loss, indicating that tissue processing may
demonstrated that half of the leukoplakias that developed in influence the results (Gao et al., 2004a,b). Since 80% of the
buccal mucosa show expression of A antigen, even though they cases showed loss of blood-group antigen expression by
histologically appear as keratinized lesions (Gao et al., 2004a). immunohistochemical staining, mechanisms other than specific
Similar A expression was found in mechanically induced loss of alleles play a role in the AB glycosyltransferase
hyperkeratinized lesions of buccal mucosa (Dabelsteen et al., expression in oral carcinomas (Gao et al., 2004b).
1975). These findings indicate that loss of antigen is not Studies of bladder cancer indicate that loss of A/B
invariably associated with hyperkeratinization. expression involves the deletion of a large chromosomal region
including the ABO locus at 9q34 1-2, which may contain one or
Mechanisms of Aberrant ABO Antigen Expressions more tumor suppressor genes (Orlow et al., 1994, 1998). The
in Oral Carcinomas investigation of oral carcinomas for alteration in this region by
A relative down-regulation of the glycosyltransferase that is microsatellite analysis found LOH or microsatellite instability
involved in the biosynthesis of A and B antigens is seen in in this region in 1/3 of the patients. These changes correlated
association with tumor development (Mandel et al., 1992; well with the lack of expression of A and B antigens in the
Orntoft et al., 1996). Studies of bladder carcinomas indicate tissue.
that events leading to loss of A transferase is related to loss of A CpG island, a stretch of GC-rich DNA sequence, is
chromosomal regions including 9q34, which is the locus for frequently located at the 5⬘ end of regulatory regions of a gene,
the ABO gene (Orlow et al., 1998). Other studies have shown extending from the promoter to the first exon region (Bird,
that the down-regulation of the ABO genes in bladder 1986). This area of genomic sequence is subject to epigenetic
carcinomas is unrelated to loss of chromosomal areas or to modifications, including DNA methylation and histone
mutations, but is related to increased cell proliferation (Orntoft acetylation and methylation, which are known to play an
et al., 1996). important role in regulating gene expression (Bird, 2002). In
Specific allelic loss occurs in oral carcinomas of blood- cancer cells, it has been shown that an increase in regional
group AO- and BO- patients. By laser capture microscopy and DNA methylation occurs in many promoter CpG islands,

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24 Dabelsteen & Gao J Dent Res 84(1) 2005

proximal constitutive promoter

encoding most of the A/B
transcripts, since an inverse
relationship was found between
promoter hypermethylation and
A/B gene expression (Iwamoto et
al., 1999; Kominato et al., 1999).
These studies were supported by
experiments in which cells were
treated with the demethylating
agent 5-aza-2⬘-deoxycytidine, a
treatment that can result in
demethylation of the ABO
promoter region and restore
transcriptional activity. The
activity of the distal promoter is
less well-understood, but appears
to be dependent on cell types
(Kominato et al., 2002).
Hypermethylation of the distal
promoter appeared not to be of
significance for expression of
blood-group antigens in either
normal or oral carcinoma tissue.
However, hypermethylation of the
A/B gene constitutive proximal
promoter region was demonstrated
in 1/3 of the cases of oral carcin-
omas (Fig. 4) (Gao et al., 2004b).
The study also showed that
hypermethylation was frequent in
hyperplastic or dysplastic epithel-
ium adjacent to tumors. A hyper-
methylation of the A/B promoter
corresponds to the lack of
expression of blood-group antigens
seen by immunohistochemical
staining. Studies have also been
undertaken to determine if A/B
expression in normal epithelium is
controlled by hypermethylation of
Figure 3. Blood group AB patient. (A) A antigen staining is strong in normal epithelium (Nor), but the promoter. In both epidermis
absent in dysplastic (Dys) and in tumor (Tum) tissue. (B,C) Antigen B staining is weak in normal, but and keratinized epithelium from
positive in dysplastic epithelium. (D) Three-day wound stained with antibodies to blood-group antigen gingiva, it was found that, although
y
A precursor (Le ). The outgrowth of the epithelium shows loss of A and B antigens and up-regulation of
the precursor Ley (see also Fig. 5). (E,F) Oral carcinoma from a blood-group AO person. A antigen
these tissues expressed little or no
staining is positive in a part of the tumor (E) and in normal epithelium (not shown), but negative in the blood-group antigen, the A/B pro-
remaining part of the tumor (F). (G) Agarose gel of the ABO gene-specific products in the above AO moter was not hypermethylated,
person. Lanes 1, 4, 7, and 10 show undigested PCR products; lanes 2, 5, 8, and 11 show Kpn I suggesting that the hypermethyl-
digested, revealing O allele; lanes 3, 6, 9, and 12 show BstE II digested, revealing A allele. Blood- ation is a specific tumor-related
group type O control and a negative control are included. In the normal epithelium, both A and O
alleles are present; the positive-stained tumor (Tumor+) shows O allele loss, whereas the negative- event (Gao et al., 2004b).
stained tumor (Tumor-) shows A allele loss. Regional hypermethylation of
chromosomes may take place in
tumors. Hypermethylation of the
A/B allele may therefore be an
resulting in a closed chromatin configuration that disables unspecific event related to hypermethylation of larger
transcription initiation of the associated genes (Baylin and chromosomal areas (Baylin and Herman, 2000).
Herman, 2000; Esteller et al., 2001). Studies of the regulatory Hypermethylation of the death-associated protein kinase gene,
mechanism of the ABO gene transcripts have demonstrated the DAPK1, which is located proximal to the ABO gene, may
presence of two promoter regions (Kominato et al., 1999). occasionally be found but in a pattern that is unrelated to
Expression of the A/B gene in epithelial cell lines has been hypermethylation of the ABO gene (Gao et al., 2004b). A
shown to be dependent on the methylation status of the gene referred to as DBCCR1 has been identified as a tumor

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J Dent Res 84(1) 2005 ABO Blood-group Antigens in Oral Cancer 25

suppressor gene at 9q32-33

(Habuchi et al., 1997). This gene
is frequently hypermethylated in
oral cancers, but a significant
correlation to hypermethylation
of the ABO promoter has not
been found (Gao et al., 2004c).
Another gene, Tuberous Sclerosis
gene TSC1, which is suggested to
be a tumor suppressor gene at
9q34.1 (Hornigold et al., 1999),
and is located very close to the
ABO gene, shows no hyper- Figure 4. Examples of methylation status of the genes at chromosome 9q33-34. (A) MS-PCR (methylation-
specific PCR) shows the hypermethylation status of ABO, TSC1, DAPK1, and DBCCR1, all of which are
methylation in oral cancer (Gao, located at chromosome 9q33-34 in 2 tumor samples (T1 and T2). M, methylated allele; U, unmethylated
2004, unpublished data) (Fig. 4). allele. For both the DAPK1 and DBCCR1, hypermethylation occurs at the same time as hypermethylation of
These findings suggest that ABO the ABO gene promoter. There is no hypermethylation of the TSC1 gene promoter, which is close to the
hypermethylation is a tumor- ABO gene. (B) Methylation-specific melting curve analysis (MS-MCA) of a tumor and normal adjacent tissue.
The curve pattern of the tumor demonstrates that, in the tumor, both methylated and unmethylated alleles are
specific event leading to loss of found; in normal epithelium, only an unmethylated allele is present.
AB expression. Knudson's two-
hit model predicts that a
phenotypic consequence of tumor
suppressor gene loss is not seen unless both alleles of a gene propensity is not clear. It is possible that blood-group A-
are inactivated in a tumor (Knudson, 2001). The fact that A/B positive and A-negative individuals may have different binding
allelic loss or LOH in the ABO region and hypermethylation properties of as-yet-unknown micro-organisms, which may
of the ABO gene promoter are frequently found in the same then influence their susceptibility to carcinoma development.
patients also suggests that the inactivation of the A/B gene is For example, extensive studies of Helicobacter pylori, which is
of significance for oral cancer development. It is impossible, responsible for gastroduodenal ulcers and eventually gastric
however, to explain all losses of A/B antigens in oral carcinoma, have shown that one of the binding mechanisms of
carcinomas by allelic loss or hypermethylation of the ABO the Helicobacter to the mucosal cells involves blood-group
promoter, which suggests that other regulatory factors outside antigen precursor structures (Ilver et al., 1998). However, at
the ABO promoter are functional in transcriptional regulation present, there are no indications of such mechanisms in oral
of the ABO gene (Kominato et al., 1999; Gao et al., 2004b). cancer development.
Experimental studies of rat colon carcinoma cells indicate
Function of the ABO Antigen that cells with A expression are resistant to apoptosis and are
Blood-group antigens can be present on key receptors strongly tumorigenic in synergenic rats (Marionneau et al.,
controlling cell proliferation, adhesion, and motility, such as 2002). Similar findings were found when cells expressed the
epidermal growth factor receptor, integrins, cadherins, and A precursor H antigen, whereas cells expressing structures of
CD-44 (Greenwell, 1997; Hakomori, 1999; Prokopishyn et shorter carbohydrates were less tumorigenic. The significance
al., 1999). The expression patterns of these various receptors of these studies in relationship to human tumor development
differ according to the type of normal epithelium and the type is difficult to interpret. Studies in the rat and the mouse have
of cancer, and therefore the role of ABH antigens in the shown that, in stratified epithelium, the expression of blood-
biology of human cancers may also vary. The function of the group antigen is opposite that seen in humans, since basal
ABO expression in normal stratified oral epithelium is cells express the full blood-group antigen A or B, whereas
unclear. Half of the population is blood group O, and 20% of spinous cells express the precursor structure H. Therefore, the
the A/B individuals do not express A/B due to their non- rat model cannot be compared with human carcinogenesis
secretor status. In the O phenotype, the A/B precursor is (Reibel et al., 1984; Mackenzie et al., 1995; Marionneau et
transformed into the difucosylated structures Leb and Le y, al., 2002).
whereas in the non-secretors, the monofucosylated structures Studies with human colon carcinoma cell lines have shown
Lea and Lex are found (Mandel et al., 1991). It is known that that loss of expression of AB glycosyltransferase can enhance
specific strains of pathogens bind to carbohydrates of the malignancy of the cell lines (Ichikawa et al., 1997). cDNA
histo-blood-group family, and that individuals devoid of encoding A transferases was transfected into a human
carbohydrates are less likely to be infected (Stapleton et al., colorectal carcinoma cell line expressing the A precursor
1992). Although there are reports suggesting that non- structures H/Ley. The investigators thus established cell lines
secretors have a higher incidence of Candida albicans that express A antigens and showed that conversion of H to A
infections, which can be explained by differences in binding inhibited Matrigel-dependent motility, a standard measure of
of the Candida to the oral mucosa, there appears to be no tumor cell motility and invasiveness (Ichikawa et al., 1997).
difference in the function of the normal stratified oral The investigators further provided evidence that the A
epithelium in any of the different blood-group phenotypes expression seems to be associated with ␣6, ␣3, and ␤1
(Johansson et al., 2000). integrins and suggested that glycosylation of the integrins may
A higher incidence of various types of carcinomas has been affect the degree of ␣6 or ␤3 interaction with ␤1 subunit,
reported in blood-group A/B individuals, but the reason for this which is known to be involved in the Matrigel motility of

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26 Dabelsteen & Gao
Figure 5. The synthetic pathway of Ley involves the formation of H from
a precursor by the action of ␣1-2 fucosyltransferase and then the
transformation of this structure to Le y by the action of ␣ 1-3
fucosyltransferase.
colon tumor cells (Ichikawa et al., 1997). The correlation
between glycosylation and cell migration is evidenced by the
finding that the motility of a melanoma cell expressing H
precursor structures could be inhibited by anti-H antibodies
(Miyake and Hakomori, 1991). Taken together, these studies
suggest that blocking of H, either by antibodies or by
elongation of the carbohydrate chains, has a strong influence
on cell motility. Since transfection per se causes artificial
overexpression of A antigens, a later study attempted to
determine if there was any difference in cell motility among
tumor cells from a cell line in which there were both blood-
group antigen A-negative and blood-group A-positive cells
(Ichikawa et al., 1998). This study showed a marked
enhancement of motility in the A-negative population as
compared with the A-positive population. It was also shown
that A glycosylation occurs on ␣3, ␣6, and ␤1 integrin
receptors, again suggesting that altered glycosylation of the
integrins alters their function (Ichikawa et al., 1998). The fact
that the blood-group antigen expression correlates with
integrin receptors was further supported by studies showing
that ABO antigens in human colon carcinoma cells were
carried on ␣3, ␤1 integrins (Prokopishyn et al., 1999). Other
experiments have supported the finding that decreased
expression of AB blood-group structures and increased
expression of the precursor Ley are related to increased cell
motility (Figs. 3D, 5). Migrating breast carcinoma cell lines
have up-regulated expression of Ley that is found mainly on
microspikes and ruffled membranes (Garrigues et al., 1994).
These structures are involved in cell migration and are known
to express clusters of integrins and associated proteins that
mediate cell adhesion and cell signaling. The relationship
between enhanced cell migration and loss of expression of AB
antigens has been clearly demonstrated in in vivo studies of
oral mucosal wound-healing in both humans and Rhesus
monkeys (Dabelsteen and Fejerskov, 1974; Mackenzie et al.,
1977). Migrating epithelial cells show loss of AB antigens and
up-regulation of precursor H and Ley (Dabelsteen et al., 1998).
One further aspect of the H/Ley expression in wounds and
tumors is the recent finding that both Le y and H possess
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J Dent Res 84(1) 2005
angiogenic activity (Halloran et al., 2000). This finding is of
course of major interest, since angiogenesis is essential for
both tumor development and wound-healing.
Thus, studies seem to indicate that the expression of A and
B antigens inhibits cell migration. Since patients with retained
A antigen on tumor cells have a better prognosis than O
patients, it can be argued that the A antigen has a protective
function that may be related to inhibition of tumor cell motility,
although the mechanisms are at present obscure.
Increased expression of sialyl-Le x , molecules closely
related to the ABO precursor molecules, is correlated with a
poor prognosis for certain types of carcinomas. The reason for
the poor prognosis is that expression of sialosyl-Lex on tumor
cells will facilitate metastasis through binding to selectin
adhesion receptors expressed on activated platelets and
endothelial cells. However, no increased expression of either
Le x , sialyl-Le x , Le a , or sialyl-Le a has been found in oral
carcinomas. Thus, these ABO-related molecules seem to play
a minor role in the spread of oral carcinomas (Renkonen et al.,
2000).
FUTURE RESEARCH
Although the above findings suggest a function for the ABO
antigen in oral mucosa, we still lack unequivocal evidence for a
putative function. Recently, the murine genomic and
complementary DNA that is equivalent to the human ABO
gene has been cloned (Yamamoto et al., 2001). This may help
to establish a mouse model system to assess the functionality of
the ABO genes in the future.
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