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Roles of E- and P-Cadherin in the Human Skin
of Dermatology, Hamamatsu University School of Medicine, 3600 Handa-cho, Hamamatsu, 431–31, Japan
of Dermatology, Graduate School of Medicine, Kyoto University, Shogoin, Sakyo, Kyoto 606 Japan
cadherin; soluble E-cadherin; cell adhesion molecules; cancer; pemphigus; cell
The Ca21-dependent cell-cell adhesion molecules, termed cadherins, are subdivided
into several subclasses. E (epithelial)- and P (placental)-cadherins are involved in the selective
adhesion of epidermal cells.
E-cadherin is expressed on the cell surfaces of all epidermal layers and P-cadherin is expressed
only on the surfaces of basal cells. Ultrastructural studies have shown that E-cadherin is distributed
on the plasma membranes of keratinocytes with a condensation in the intercellular space of the
desmosomes. During human skin development P-cadherin expression is spatiotemporally controlled
and closely related to the segregation of basal layers as well as to the arrangement of epidermal cells
into eccrine ducts. In human skin diseases E-cadherin expression is markedly reduced on the
acantholytic cells of tissues in pemphigus and Darier’s disease.
Cell adhesion molecules are now considered to play a significant role in the cellular connections of
cancer and metastatic cells. Reduced expression of E-cadherin on invasive neoplastic cells has been
demonstrated for cancers of the stomach, liver, breast, and several other organs. This reduced or
unstable expression of E- and P-cadherin is observed in squamous cell carcinoma, malignant
melanoma, and Paget’s disease, but cadherin expression is conserved in basal cell carcinoma.
Keratinocytes cultured in high calcium produce much more intense immunofluorescence of
intercellular E- and P-cadherin than those cells grown in low calcium. E-cadherins on the plasma
membrane of the keratinocytes are shifted to desmosomes under physiological conditions, and
therein may express an adhesion function in association with other desmosomal cadherins.
Soluble E-cadherins in sera are elevated in various skin diseases including bullous pemphigoid,
pemphigus vulgaris, and psoriasis, but not in patients with burns. Markedly high levels in soluble
E-cadherin are demonstrated in patients with metastatic cancers. Microsc. Res. Tech. 38:343–352,
1997. r 1997 Wiley-Liss, Inc.
Cell adhesion mechanisms play a major role in vital
processes such as embryogenesis, tissue and organ
pattern formation, and maintenance of specific tissue
architecture. Alternations in cell adhesion molecules
(CAM) have been implicated in the loss of control of cell
proliferation and neoplasia. Various classes of cell-cell
adhesion molecules are differentially expressed in the skin
during development and epidermal cell-cell adhesion is
involved, directly or indirectly, in the induction process.
Cadherins, which are transmembranous polypeptides that connect one cell to another through calcium
ion-dependent homophilic binding (Nagafuchi et al.,
1987; Nose et al., 1988; Takeichi, 1988; Takeichi, 1991),
are subdivided into several subclasses such as E (epithelial)-cadherin, N (neural)-cadherin, P (placental)cadherin, and L-CAM (liver cell adhesion molecule).
Using genetic technology, more than 20 cadherins have
been described in the central nervous system, on liver
and vascular endothelial cells and in other tissues and
organs (Matsuyoshi, 1993a; Suzuki et al., 1991). Thus,
the cadherin family of adhesion molecules is much
larger than previously thought.
Recent studies on cadherins have focused on cytoplasmic adhesion mechanisms. Sequence comparison of the
classic cadherins and the heterotypic expression studies of mutant proteins led to the identification of
cytoplasmic proteins of 102, 88, and 80 kDa, which
complex with cadherins via a specific cytoplasmic binding domain. These proteins, termed a-, b-, and g-catenin
respectively, are crucial for cadherin function. Catenins
link cadherins to the actin filamentous network and to
other transmembrane and cytoplasmic proteins.
a-catenin is homologous to vinculin, and b-catenin is
homologous to the product of the Drosophila gene
armadillo, while g-catenin seems to be identical to
plakoglobin. Structural and functional analyses of catenins have indicated that cytoplasmic anchorage is important in mediating adhesive interactions. In addition,
the anchorage of the cadherin-catenin complex to the
cytoskeleton possibly depends upon tyrosine phosphorylation (Kemler, 1993; Takeichi et al., 1992).
*Correspondence to: Fukumi Furukawa, MD, Department of Dermatology,
Hamamatsu University School of Medicine, 3600 Handa-cho, Hamamatsu,
431–31, Japan.
Received 18 November 1994; Accepted in revised form 16 March 1995
TABLE 1. Classification of the cadherin superfamily
Cadherins are essential for the mutual association of
vertebrate cells, and through their homophilic binding
interactions they play a role in cell-sorting mechanisms
and confer adhesion specificities to cells (Takeichi,
1991; Larue et al., 1994). Cadherins are thus considered to be important regulators of morphogenesis in
various organs, including the skin. Cadherins present
in the epidermal cells are involved not only in maintaining the arrangement of these cells but also in dermal
condensation (Hirai et al., 1989). Recent pathological
examinations suggest that the down-regulation of cadherin expression is associated with increased invasiveness of tumor cells (Birchmeier et al., 1993). Furthermore, the role of soluble E-cadherin is the subject of
much discussion (Katayama et al., 1994).
In this review, we describe the roles of E- and
P-cadherins in human skin development, cultured keratinocytes, and skin cancers.
ies on the pathogenesis of autoimmune bullous diseases, especially pemphigus (Amagai, 1995; Amagai et
al., 1991; Stanley, 1995). The second family of cadherins
has multi-repeated extracellular domains and unique
intracellular domains. The third family has the enzyme
activity of tyrosine phosphorylation.
Other classifications schemes for the cadherin family
have been proposed. Pouliot (1992) reported the result
of phylogenic analyses of the cadherin superfamily. He
studied the first extracellular domain of cadherin which
is the site responsible for mediating adhesive interaction, and showed that cadherins can be classified into
three groups with distinct structural features. Group I
includes N-, E-, P-, B(brain)-, R(retinal)-cadherin, and
L-CAM(liver cell adhesion molecule). Group II is
M(muscle)-cadherin. Group III is T(truncated)-cadherin. Phylogenically, N-cadherin shows very little
sequence divergence between species, whereas all other
cadherin subtypes show more substantial divergence,
suggesting that selective pressure upon the first domain may be greater for N-cadherin than for the other
cadherins. The protocadherin family, which is characterized by a distinct cadherin repeat in the extracellular
domain, was proposed by Sano et al. (1993).
Rimm and Morrow (1994) pointed out the relatedness
of different cadherins based on the molecular cloning of
human E-cadherin. The gene encoding full-length human E-cadherin has been cloned and sequenced from
liver and colon cDNA libraries. The derived amino acid
sequence of the extracellular domain of human Ecadherin displays strong homologies with all other
vertebrate cadherins, including the presence of the EC
(extracellular) 1 domain. The sequence of the cytoplasmic domain defines it as a particular subtype of cadherin. In addition, the cytoplasmic domain contains two
distinct <30–35 amino acid sequence motifs that are
conserved in several but not all cadherins. Referring to
these motifs as cadherin homology domains ‘‘two’’ (CH2)
and ‘‘three’’ (CH3), the authors suggest a sub-classification of the cadherins that is cognizant of the divergence
in the cytoplasmic domain, and identify two sequence
motifs that may represent functional cassettes variably
used in some of the cadherins and possibly other
proteins as well.
A classification of the cadherin superfamily, based on
their homology of extracellular domains (Table 1), was
proposed by Matsuyoshi (1993). The first family, which
has 4-repeated extracellular domains, includes the
classic cadherin, desmosomal cadherins, and cadherins
without intracellular domains. Classic cadherins can be
subdivided further into adherens junction cadherin,
unidentified cadherin, and several others. The E- and
P-cadherins of adherens junction cadherin are expressed on cell surfaces of the epidermis. Cadherin 5
localized on human and mouse vascular endothelial
cells, provides new insights into vascular morphogenesis and regulation (Matsuyoshi et al., 1997). Desmosomal cadherins include pemphigus vulgaris antigen
(PVAg, desmoglein III), desmoglein I, II, and desmocollin I, II, III (Amagai et al., 1991; Buxton et al., 1993;
Collins et al., 1991; Koch et al., 1990). Identification of
these desmosomal cadherins opened the door for stud-
Among these cadherins, E- and P-cadherins have
been investigated thoroughly in relation to mouse skin
morphogenesis. In mouse skin, E-cadherin is expressed
in all cells of the embryo at the implantation stage.
During development, E-cadherin disappears from some
cell layers in the mesoderm. In older embryos, essentially all proliferating epithelial cells derived from the
ectoderm and the endoderm express E-cadherin, although some terminally differentiated and/or keratinized cells lose E-cadherin. P-cadherin was first detected
in the extraembryonic layers of early embryos, such as
the ectoplacental cone and the visceral endoderm, at
the stage of implantation. Around the neurula stage,
various tissues begin to express P-cadherin. In the fetal
epidermis, P- and E-cadherin are differentially expressed; the germinative basal layer has P-cadherin as
A. 4-repeated family
1. Classic cadherin
(1) Adherens junction cadherin
E-cadherin (uvomorulin)
(2) Unidentified localization sites
cadherin 6, 8, 9, 10, 11
(3) Others
M-cadherin (muscle cadherin)
Cadherin 5 (VE-cadherin) (endothelial cadherin)
2. Cadherins without intracytoplasmic domains
3. Desmosome cadherin
desmolgein I, II, III
desmocollin I, II, III (IV)
B. Multi-repeated cadherin
Fat, PC42, PC43
C. Others
This classification is based on the report of Matsuyoshi (1993) and Buxton et al.
Fig. 1. Photographs of immunofluorescence localization of E- and
P-cadherins in the sole of a 5-year-old infant. Specimens were fixed in
periodate-lysine-paraformaldehyde (PLP) solution and 6 µm cryosectioned specimens were incubated with a monoclonal antibody (mAb) to
human E-cadherin (HECD-1) or a mAb to human P-cadherin (NCCCAD-299). FITC-conjugated goat anti-mouse IgG was used as the
second antibody. Detailed procedures are described in our previous
well as E-cadherin and the middle layer has only
E-cadherin (Takeichi, 1988). As for the hair, P-cadherin
is expressed in proliferating regions such as the basal
layer, the outer root sheath, and the hair matrix,
whereas E-cadherin is expressed in both proliferating
and non-proliferating regions (Hirai et al., 1989).
In human adult and infant skin, E-cadherin is expressed on the cell surfaces of all epidermal layers
including skin appendages, whereas P-cadherin is expressed only on cells of the basal layers and the outer
layers of skin appendages (Fig. 1). These latter observations suggest that P-cadherin is associated with the
proliferating cell compartment (Fujita et al., 1992).
Ultrastructurally, E-cadherin is distributed on all of the
plasma membranes of keratinocytes, but not on the
dermal surface of basal cells. Dense deposits of Ecadherin are found in the intercellular space of desmosomes (Fig. 2), and P-cadherin is localized on the
surfaces of basal cells, but not on the dermal surface
(Horiguchi et al., 1994). In human organs E-cadherin is
expressed in almost all epithelial tissues, while the
distribution of P-cadherin is restricted to the basal or
lower layers of stratified epithelia in which both cadherins are co-expressed (Shimoyama et al., 1989).
In human fetal skin, the patterns of E- and Pcadherin expression are generally similar to those
found in the adult (Fujita et al., 1992; Furukawa et al.,
1989). Embryonic skin at 7–8 weeks of estimated
gestational age (EGA) consists of basal and peridermal
cell layers. Basal cells have both E- and P-cadherin on
their surfaces, whereas some of the periderm cells have
only E-cadherin. After 9 weeks of EGA the intermediate
cells have only E-cadherin, and the periderm cells have
neither of the cadherins. When hair follicles develop
after 12 weeks of EGA their associated structures—the
outer root sheath, the outer-most layer of the hair
matrix, and the sebaceous gland—express both E- and
P-cadherins. P-cadherin temporarily shows a unique
spatiotemporal expression pattern in the developing
report (Reproduced with permission from Fujita, M., Furukawa, F.,
Fujii, K., Horiguchi, Y., Takeichi, M., and Imamura, S. (1992) Expression of cadherin cell adhesion molecules during human skin development: morphogenesis of epidermis, hair follicles and sweat ducts.
Arch. Dermatol. Res., 284:159–166. rSpringer-Verlag GmbH & Co.
KG). Bar: 40 µm.
Fig. 2. Photograph of ultrastructural localization of E-cadherin in
adult skin (38,000). Samples were fixed in PLP solution and cryosections (6 µm) were allowed to react with HECD-1. The sections were
incubated with biotinylated anti-mouse IgG and then with soluble
avidin-biotin-peroxidase complex. Other details were described recently (Horiguchi et al., 1994). BC: basal cell, De: dermis. An arrow
indicates E-cadherin on the desmosome. Bar: 1 µm.
sweat ducts. During this stage, P-cadherin accumulates on cells in the epidermal ridges that are distinct
from the proliferating cells detected by Ki-67. The
expression of P-cadherin is spatiotemporally controlled,
and may be closely related to the segregation of the
basal layers and the arrangement of epidermal cells
into eccrine sweat ducts.
Fig. 3. Immunofluorescence photograph of E-cadherin localization in an early lesion of pemphigus
vulgaris. Reduced E-cadherin expression is observed at the acantholytic cells. Bar: 40 µm.
Autoimmune Bullous Diseases
Of special interest is the E-cadherin expression pattern in skin diseases since E-cadherin plays an essential role in cell-cell adhesion. The desmosome cadherins
listed in (Table 1) play essential roles in the pathogenesis of autoimmune bullous diseases. E-cadherin, one of
the adherens junction cadherins, accumulates at desmosomes and exhibits, in vivo and in vitro, the distribution
pattern described above (Horiguchi et al., 1994).
In early lesions of pemphigus, the immunofluorescence intensity of E-cadherin is markedly reduced
especially on the acantholytic cells, whereas at the
non-acantholysis sites E-cadherin shows an intercellular distribution with a near uniform immunofluorescence intensity (Fig. 3) (Furukawa et al., 1990; Furukawa et al., 1989). Similar staining patterns were
observed in our previous report of Darier’s disease
(Furukawa et al., 1991), for which the gene mapped to
chromosome 12q (Bashir et al., 1993; Buxton, 1993;
Craddock et al., 1993). However, the fully developed
bullae in PV show an intercellular distribution of
E-cadherin with a near uniform immunofluorescence
intensity for this molecule in both the suprabasal cell
layer and the separated roof epidermis (Furukawa et
al., 1994). The epitope(s) of E-cadherin was considered
to be different from those of pemphigus vulgaris antigens (PVA) (Furukawa et al., 1989).
Amagai, Stanley and co-workers (1991) cloned the
cDNA encoding pemphigus vulgaris antigen (PVA) using antibodies obtained from patients. They demonstrated by sequence analysis that PVA belongs to the
cadherin family. PVA is now recognized as desmoglein
III, but PVA lacks the biochemical properties characteristic of classical cadherins (Plott et al., 1994). Differences are observed in catenin binding and protection of
the extracellular domain by calcium from degradation
by trypsin. These results suggest that PVA and classical
cadherins may not subserve identical biological functions.
Skin Cancers
Cancers are characterized by their non-regulated cell
growth, loss of polarity, and invasion or metastasis.
Several recent studies have suggested that the loss of
E-cadherin may be associated with tumor progression
and E-cadherin acts principally as a suppressor of
invasive ability (Behrens et al., 1989; Birchmeier et al.,
1993; Navarro et al., 1991). In fact, reduced amounts of
cadherins have been immunohistochemically described
in human cancers from various organs such as esophagus, stomach, liver, breast, head, neck, prostate, and
gingiva. Several mechanisms of impaired function of
cadherin are now hypothesized (Fig. 4) (Matsuyoshi,
1993; Takeichi, 1991).
In murine skin squamous cell carcinoma induced by
chemical carcinogens, the association between cadherin expression and tumor progression has been investigated by Cano and colleagues (Navarro et al., 1991).
They studied the expression of E- and P-cadherin in
various mouse epidermal keratinocyte cell lines that
are representative of different stages of epidermal
carcinogenesis. Complete absence of E-cadherin mRNA
and protein was found in the epithelioid type and
fibroblastoid type of epidermal keratinocyte cell lines,
which all grew as fast as tumors in nude mice. In
contrast, well differentiated squamous cell carcinoma
expressed considerable amounts of E-cadherin. Pcadherin was detected in all cell lines irrespective of cell
differentiation. The introduction of an exogeous Ecadherin cDNA into epithelioid type cells leads to a
partial suppression of the tumorigenicity. This finding
Fig. 4.
Proposed mechanisms of cadherin dysfunction.
suggests that E-cadherin expression is associated with
the inhibition of tumor growth in some cases of experimentally induced mouse skin carcinomas. Reduced
E-cadherin expression in aggressive squamous cell
carcinoma is suggested to be associated with a redistribution or absence of sodium, potassium-adenosine triphosphatase (Na1, K1-ATPase), a marker of cell polarity (Ruggeri et al., 1992), in the combined function of
catenin complex (Nathke et al., 1994) and membranecytoskeleton (Marrs et al., 1993).
Impaired cadherin functions have been proposed in
human skin cancers. For example, the transformed
keratinocyte cell line HSC-1 (Kondo and Aso, 1981),
originally derived from human squamous cell carcinoma shows a reduced and dot-like distribution of
E-cadherin (Furukawa et al., 1994). E-cadherin expression by basal cell carcinoma (BCC) is preserved in
superficial and nodular types but reduced in infiltrative
BCC, which suggests E-cadherin is related to the
growth pattern and the local aggressive behaviour of
BCC (Pizarro et al., 1994). A lower or unstable expression of E-cadherin is also found in squamous cell
carcinoma and Paget’s disease (Shirahama et al., 1996).
Expressions of E- and P-cadherins are correlated with
the degree of differentiation of squamous cell carcinoma
and a marked reduction of E-cadherin is observed with
the appearance of metastatic squamous cell carcinoma
cells in lymph nodes (Furukawa et al., 1994; Shirahama
et al., 1996). In addition, immunohistochemical studies
indicate that epidermal cells of solar keratosis show
unstable expression of cadherins. The expression of
P-cadherin in malignant melanoma is associated with
tumor proliferation and progression. These findings
support the hypothesis that E-cadherin and P-cadherin
are involved in invasion or metastasis in vivo.
The significance of tyrosine phosphorylation in the
action of E-cadherin has also been intensely investigated. V-src oncogene is a potent effector of epithelial
differentiation and invasiveness, and promotes the
dedifferentiation through phosphorylation of the Ecadherin/catenin complex (Behrens et al., 1994). Matsuyoshi et al. (1992) observed that direct or indirect
v-src-mediated tyrosine phosphorylation perturbs cadherin function in metastatic fibroblasts. In addition,
they demonstrated that the inhibition of tyrosine phosphorylation restores cadherin action to its normal state.
Mutations of the E-cadherin gene in human gastric
carcinoma cell lines were recently reported (Oda et al.,
1994; Becker et al., 1994). E-cadherin-mediated cellcell adhesion potential may be hampered by mutations
in the structural region of the E-cadherin gene. Similar
approaches need to be applied to skin cancers, and the
contribution of other CAM family molecules to cancer
should be investigated.
Based on experimental data, E- and P-cadherin are
now considered to be key adhesion molecules in the
primary event-neoplastic cell detachment from tumor
mass. For the next step in metastasis formation, neoplastic cells have to cope with the basement membrane,
which requires digestion of and interaction with extracellular matrix components. Both of these interactions
involve the integrin family in association with altered
cadherin function (Brichmeier and Behrens, 1994).
It is well known that calcium switching induces
keratinocyte differentiation or stratification. When keratinocytes are cultured in low Ca21 medium, they do
not form any desmosomes. However, once they are
transferred to normal Ca21 medium, they start to make
cell-cell contact and to reproduce desmosomes. In controlling the stratification of keratinocytes, the adherens
junctions, like desmosomes, play an essential role
(O’Keefe et al., 1987). Thus, since cadherin function is
calcium dependent, cadherins are one of the keys in
controlling keratinocyte differentiation or stratification.
Normal human keratinocytes (NHK), cultured to
50–60% confluency in low Ca21 medium (0.03 mM),
show no obvious cell-cell interaction and only faint
localization of E-cadherin. NHK in 0.6 mM Ca21containing medium show a well defined meshwork
pattern of intercellular distribution of E-cadherin (Fig.
5) (Furukawa et al., 1990; Furukawa et al., 1994;
Horiguchi et al., 1994). At 90–100% confluency, after
culturing with high Ca21 (1.0 mM), a uniform distribution of E-cadherin is observed and its expression is
much higher than that seen with low Ca21. In contrast,
the expression of P-cadherin is reduced in high Ca21
medium (Furukawa et al., 1994). Ultrastructural studies reveal that E-cadherin distributes all around the
free surface of the plasma membrane of NHK in low
Ca21 medium, whereas high Ca21 induces its accumulation at desmosomes (Fig. 6) (Horiguchi et al., 1994).
With an immunoblot analysis no apparent differences
Fig. 5. Immunofluorescence micrographs showing E-cadherin localization in normal human cultured keratinocytes in KGM (keratinocyte
growth medium) with 0.6 mM (A) and 0.1 mM (B) calcium. Cultured
keratinocytes were fixed in acetone, incubated with diluted HECD-1
and then reacted with FITC-conjugated anti-mouse IgG. Bar: 50 µm.
(Reproduced, with permissions, from Horiguchi, Y., Furukawa, F.,
Fujita, M., and Imamura, S.: Ultrastructural localization of Ecadherin cell adhesion molecule on the cytoplasmic membrane of
keratinocytes in vivo and in vitro, Journal of Histochemistry and
Cytochemistry, 42:1333–1340, 1994).
are seen in the expression pattern of E-cadherin in
NHK cultured in the various Ca21 concentrations (Fig.
7). These results suggest that under physiological conditions E-cadherin on the plasma membrane of NHK
shifts to the desmosomes where it expresses an adhesion function.
Recently Hodivala and Watt (1994) reported that
cadherins play a role in the down-regulation of integrin
expression that occurs during keratinocyte terminal
differentiation. Thus, a cadherin-catenin complex and
other cell adhesion molecules appear to be involved in
keratinocyte differentiation.
Cadherin functions have been assayed using cell
aggregation assays (Takeichi, 1977). Briefly, cell suspensions, harvested by trypsin solution in the presence of
calcium, are incubated to allow cell aggregation on a
gyratory shaker, and then the degree of cell aggregation
is determined and expressed by the aggregation index.
Using this aggregation assay the generation of cell-cell
contacts can be determined. Cells treated with purified
pemphigus vulgaris antibody showed much lower aggregation rates than cells treated with purified normal
human IgG (Fig. 8). Pemphigus vulgaris antibody
produced an extra band of E-cadherin in Western blot
analysis (Takeichi et al., 1990). These results suggest
that pemphigus vulgaris antibody possibly induces the
degradation of E-cadherin through protease action. It is
well known that in the process of blister formation in
pemphigus vulgaris, induction of plasminogen activator synthesis, secretion of plasminogen activator, and
activation of plasminogen to plasmin occur (Hashimoto
et al., 1983). These reactions might trigger the dysfunction of E-cadherin and/or P-cadherin.
E-cadherin is involved in the persistence of Langerhans cells in epidermis (Tang et al., 1993). Fresh
murine Langerhans cells express cadherins and Langerhans cells adhere to keratinocytes in vitro through
E-cadherin. Cultured Langerhans cells express lower
levels of E-cadherin and exhibit decreased affinity for
keratinocytes. These findings suggest that E-cadherin
mediates adhesion of epidermal Langerhans cells to
A monoclonal anti-E-cadherin antibody slightly reduces the cell-cell contact of cultured NHK and also the
increases intracytoplasmic Ca21 concentration of these
cells under certain conditions (Wakita et al., manuscript in preparation). Anti-E-cadherin plus anti-Pcadherin inhibit the formation of adherens junctions
Fig. 6. Immunoelectron microscopic view of normal human cultured keratinocytes in 0.1 mM calcium (A,B) and 0.6 mM calcium
(C,D) calcium containing medium. Cultured cells on LabTek chamber
slides were washed with PBS supplemented with the same calcium
concentration, and fixed with the PLP solution. Cells were incubated
with diluted HECD-1 using the immunoperoxidase method. Resinembedded immunoperoxidase-stained cultured cells were detached
from the glass slides. Details were given in the report of Horiguchi et
al. (1994). Fig. A & C: vertical sections, Fig. B & D: horizontal sections,
Kc: keratinocyte, arrow heads or arrows: E-cadherin. Bars: A,C 5 1
µm; B,D 5 0.1 µm. (Reproduced, with permissions, from Horiguchi, Y.,
Furukawa, F., Fujita, M., and Imamura, S., Ultrastructural localization of E-cadherin cell adhesion molecule on the cytoplasmic membrane of keratinocytes in vivo and in vitro. Journal of Histochemistry
and Cytochemistry, 42:1333–1340, 1994).
and desmosomes, prevent reorganization of the cytoskeleton, and block stratification (Lewis et al., 1994).
Cadherin function is required for calcium-induced intercellular junction organization. Balda et al. (1993) found
a promotive role for diacylglycerol in the assembly of
the tight junctions of MDCK cells. These results suggest the importance of activated signaling pathways in
E-cadherin-mediated cell-cell adhesion.
Soluble E-cadherins were first identified as 80–84
kDa peptides released from MCF-7 human carcinoma
cells (Damsky et al., 1991). These peptides retain their
functional activities of disrupting cell-cell contact in
cultured epithelial cells, and antibodies against them
induce disruption of the mutual adhesion of target cells
Fig. 7. Immunoblot analysis of E- and P-cadherins in normal
human cultured keratinocytes in KGM supplemented with various
calcium concentration. Detailed procedures were based on the report
of Matsuyoshi et al. (1992).
(Wheelock et al., 1987; Wheelock and Jensen, 1992).
sE-cadherin is probably a degradation product of the
120 kDa form of intact E-cadherin and is generated by a
Ca21-dependent proteolytic action. Thus, sE-cadherin
should be a good indicator for monitoring the regeneration of E-cadherin in vivo.
sE-cadherin is elevated in several skin diseases
including bullous pemphigoid, pemphigus vulgaris, psoriasis, and other inflammatory diseases (Matsuyoshi et
al., 1995), but not in collagen diseases or in burn
patients. The elevated levels of sE-cadherin in autoimmune bullous diseases decrease after therapy. The
blister fluids of bullous pemphigoid patients contain
two times more sE-cadherin than is found in their
serum, which itself is twice as high as the levels in
normal serum. The levels of sE-cadherin in skin lesions
correlate with the levels in the circulation in both
normal controls and bullous pemphigoid patients. In
psoriasis there is a weak, but significant, association
between sE-cad levels and PASI score, which suggests
that dysregulation of epidermal cell growth might be
related to the increase of sE-cadherin. In cases of skin
cancer, several patients showed elevated levels of sEcadherin (Furukawa et al., 1994). Such patients with
squamous cell carcinoma, Paget’s disease and malignant melanoma had metastasis or widely distributed
lesions. Katayama et al. (1994) reported increased
circulating levels of sE-cadherin in patients with gastric carcinoma or hepatocellular carcinoma. Since sE-
Fig. 8. Aggregation assay of E-cadherin function using cultured
transformed human epidermal cells (HSC-1) treated with pemphigus
vulgaris antibody. Cells were incubated with purified IgG (5–10
mg/ml) for 8 or 24 hours. The culture medium was DMEM supplemented with 1% bovine serum albumin. IgG was purified with a
protein A sepharose affinity column. IgG-treated cell suspensions were
obtained with 0.01% trypsin in PBS supplemented with 1 mM calcium,
and then prepared for the cell aggregation assay. The results are
expressed as the aggregation index. X axis represents the incubation
time. A higher index indicates a lower binding ability of cadherin. The
basic protocol is similar to that reported by Matsuyoshi et al. (1992).
cadherin disrupts cell-cell junctions in vitro (Wheelock
et al., 1987), it is thought that the presence of sEcadherin up-regulates the invasiveness of tumor cells.
These results suggest that sE-cadherin is a useful
marker of various diseases. Although the role and
origin of sE-cadherin remain controversial, the elevated levels of sE-cadherin would inhibit cell-cell adhesion and thus favor bullous formation or carcinoma
Cadherins are involved in vital processes such as
embryogenesis, tissue and organ pattern formation,
and maintenance of specific organ architecture. Cadherin dysfunction may induce various skin diseases
including bullous diseases and metastatic or invasive
cancers. Investigations into the family of cadherin
molecules will provide new and exciting insight into
cutaneous biology. The focus in cadherin research has
recently changed from extracellular binding to cytoplasmic control of cell adhesions by the cadherin-catenin
complex. We believe that in the near future the control
systems regulating cadherin molecules will be more
clearly and completely understood.
This work was supported by grants from the Japanese Ministry of Welfare and Health, and the Japanese
Ministry of Science, Culture and Education.
Amagai, M. (1995) Adhesion molecules. 1. keratinocyte-keratinocyte
interactions; cadherins and pemphigus. J. Invest. Dermatol., 104:
Amagai, M., Kalus-Kovtun, V., and Stanley, J.R. (1991) Autoantibodies
against a novel epithelial cadherin in pemphigus vulgaris, a disease
of cell adhesion. Cell, 67:869–877.
Balda, M.S., Gonzalez-Mariscal, L., Matter, K., Cereijido, M., and
Anderson, J.M. (1993) Assembly of the tight junction: the role of
diacylglycerol. J. Cell Biol., 123:293–302.
Bashir, R., Munro, C.S., Mason, S., Stephenson, A., Rees, J.L., and
Strachan, T. (1993) Localisation of a gene for Darier’s disease. Hum.
Molecular Genet., 2:1937–1939.
Becker, K-F., Atkinson, M.J., Reich, U., Becker, I., Nekarda, H.,
Siewert, J.R., and Hofler, H. (1994) E-cadherin gene mutations
provide clues to diffuse type gastric carcinomas. Cancer Res.,
Behrens, J., Mareel, M.M., van Roy, F.M., and Birchmeier, W. (1989)
Dissecting tumor cell invasion: epithelial cells acquire invasive
properties after the loss of uvomorulin-mediated cell-cell adhesion.
J. Cell Biol., 108:2435–2447.
Behrens, J., Vakaet, L., Friis, R., Winterhager, E., Roy, F.V., Mareel,
M.M., and Birchmeier, W. (1993) Loss of epithelial differentiation
and gain of invasiveness correlates with tyrosine phosphorylation of
the E-cadherin/b-catenin complex in cells transformed with a temperature-sensitive v-src gene. J. Cell Biol., 120:757–766.
Birchmeier, W., and Behrens, J. (1994) Cadherin expression in carcinomas: role in the formation of cell junctions and the prevention of
invasiveness. Biochim. Biophys. Acta, 1198:11–26.
Birchmeier, W., Weidner, K.M., Hulsken, J., and Behrens, J. (1993)
Molecular mechanisms leading to cell junction (cadherin) deficiency
in invasive carcinomas. Semi. Cancer Biol., 4:231–239.
Buxton, R.S. (1993) Yet another skin defect, Darier’s disease, maps to
chromosome 12q. Hum. Molecular Genet., 2:1763–1764.
Buxton, R.S., Cowin, P., Franke, W.W., Garrod, D.R., Green, K.J.,
King, I.A., Koch, P.J., Magee, A.I., Rees, D.A., Stanley, J.R., and
Steinberg, M.S. (1993) Nomenclature of the desmosomal cadherins.
J. Cell Biol., 121:481–483.
Collins, J.E., Legan, P.K., Kenny, T.P., MacGarvie, J., Holton, J.L., and
Garrod, D.R. (1991) Cloning and sequence analysis of desmosomal
glycoprotein 2 and 3 (desmocollins): cadherin-like desmosomal
adhesion molecules with heterogenous cytoplasmic domains. J. Cell
Biol., 113:381–391.
Craddock, N., Dawson, E., Burge, S., Parfitt, L., Mant, B., Roberts, Q.,
Daniels, J., Gill, M., McGuffin, P., Powell, J., and Owen, M. (1993)
The gene for Darier’s disease maps to chromosome 12q23-q24.1.
Hum. Molecular Genet., 2:1941–1943.
Damsky, C.H., Richa, J., Solter, D., Knudsen, K., and Buck, C.A. (1991)
Identification and purification of a cell surface glycoprotein mediating intercellular adhesion in embryonic and adult tissue. Cell,
Fujita, M., Furukawa, F., Fujii, K., Horiguchi, Y., Takeichi, M., and
Imamura, S. (1992) Expression of cadherin cell adhesion molecules
during human skin development: morphogenesis of epidermis, hair
follicles and sweat ducts. Arch. Dermatol. Res., 284:159–166.
Furukawa, F., Fujii, K., Matsuyoshi, N., Horiguchi, Y., and Imamura,
S. (1990) The cadherins: cell-cell adhesion molecules possibly control pathogenesis of pemphigus. J. Invest. Dermatol., 94:526 (Abstract).
Furukawa, F., Kanauchi, H., and Imamura, S. (1989) Localization of
E-cadherin in human skin tissue. Conn. Tissue, 21:3–4.
Furukawa, F., Miyachi, Y., Wakai, Y., and Imamura, S. (1991) Localization of cadherin cell-adhesion molecules in the skin lesion of Darier
disease. Int. J. Dermatol., 30:599–600.
Furukawa, F., Takigawa, M., Matsuyoshi, N., Shirahama, S., Wakita,
H., Fujita, M., Horiguchi, Y., and Imamura, S. (1994) Cadherins in
cutaneous biology. J. Dermatol., 21:802–813.
Hashimoto, K., Shafran, K., Webber, P., Lazarus, G.S., and Singer,
K.H. (1983) Anti-cell surface pemphigus autoantibody stimulates
plasminogen activator activity of human epidermal cells: a mecha-
nism for the loss of epidermal cohesion and blister formation. J. Exp.
Med. 157:259–272.
Hirai, Y., Nose, A., Kobayashi, S., and Takeichi, M. (1989) Expression
and role of E- and P-cadherin adhesion molecules in embryonic
histogenesis. II. skin morphogenesis. Development, 105:271–277.
Hodivala, K.J., and Watt, F.M. (1994) Evidence that cadherins play a
role in the downregulation of integrin expression that occurs during
keratinocyte terminal differentiation. J. Cell Biol., 124:589–600.
Horiguchi, Y., Furukawa, F., Fujita, M., and Imamura, S. (1994)
Ultrastructural localization of E-cadherin cell adhesion molecule on
the cytoplasmic membrane of keratinocytes in vivoand in vitro. J.
Histochem. Cytochem., 42:1333–1340.
Katayama, M., Hirai, S., Kamihagi, K., Yasumoto, M., and Kato, I.
(1994) Soluble E-cadherin fragments increased in circulation of
cancer patients. Br. J. Cancer, 69:580–585.
Kemler, R. (1993) From cadherins to catenins: cytoplasmic protein
interactions and regulation of cell adhesion. T.I.G., 9:317–321.
Koch, P.J., Walsh, M.J., Schmelz, M., Goldschmidt, M.D., Zimbdmann,
R., and Franke, W.W. (1990) Identification of desmoglein, a constitutive desmosomal glycoprotein, as a member of the cadherin family of
cell adhesion molecules. Eur. J. Cell Biol., 53:1–12.
Kondo, S., and Aso, K. (1981) Establishment of a cell line of human
skin squamous cell carcinoma in vitro. Br. J. Dermatol., 105:125–
Larue, L., Ohsugi, M., Hirchenhain, J., and Kemler, R. (1994) Ecadherin null mutant embryos fail to form a trophectoderm epithelium. Proc. Natl. Acad. Sci., USA, 91:8263–8267.
Lewis, J.E., Jensen, P.J., Wheelock, M.J. (1994) Cadherin function is
required for human keratinocytes to assemble desmosomes and
stratify in response to calcium. J. Invest. Dermatol., 102:870–877.
Marrs, J.A., Napolitano, E.W., Murphy-Erdosh, C., Mays, R.W., Reichardt, L.F., and Nelson, W.J. (1993) Distinguishing roles of the
membrane-cytoskeleton and cadherin mediated cell-cell adhesion in
generating different Na1, K1-ATPase distribution in polarized epithelia. J. Cell Biol., 123:149–164.
Matsuyoshi, N. (1993a) Cadherin family and adhesion in cancer cells.
Biotherapy, 7:1158–1165 (in Japanese).
Matsuyoshi, N., Hamaguchi, M., Taniguchi, S., Nagafuchi, A., Tsukita,
S., and Takeichi, M. (1992) Cadherin-mediated cell-cell adhesion is
perturbed by v-src tyrosine phosphorylation in metastatic fibroblasts. J. Cell Biol., 118:703–714.
Matsuyoshi, N., Toda, K-I., Horiguchi, Y., Tanaka, T., Nakagawa, S.,
Takeichi, M., and Imamura, S. (1997) In vivo evidence of the critical
role of cadherin-5 in murine vascular integrity. Proc. Ass. Am.
Physicians 109: (in press)
Matsuyoshi, N., Tanaka, T., Toda, K-I., Okamoto, H., Furukawa, F.,
and Imamura, S. (1995) Soluble E-cadherin: a novel cutaneous
disease marker. Br. J. Dermatol, 132:745–749.
Nagafuchi, A., Shirayoshi, Y., Okazaki, K., Yasuda, K., and Takeichi,
M. (1987) Transformation of cell adhesion properties by exogenously
introduced E-cadherin cDNA. Nature, 329:341–343.
Nathke, I.S., Hinck, L., Swedlow, J.R., Papkoff, J., and Nelson, W.J.
(1994) Defining interactions and distributions of cadherin and
catenin complexes in polarized epithelial cells. J. Cell Biol., 125:1341–
Navarro, P., Gomez, M., Pizarro, A., Gamallo, C., Quintanilla, M., and
Cano, A. (1991) A role for the E-cadherin cell-cell adhesion molecule
during tumor progression of mouse epidermal carcinogenesis. J. Cell
Biol., 115:517–533.
Nose, A., Nagafuchi, A., and Takeichi, M. (1988) Expressed recombinant cadherins mediate cell sorting in model systems. Cell 54:993–
Oda, T., Kanai, Y., Oyama, T., Yoshiura, K., Shimoyama, Y., Birchmeier, W., Sugimura, T., and Hirohashi, S. (1994) E-cadherin gene
mutations in human gastric carcinoma cell lines. Proc. Natl. Acad.
Sci. USA, 91:1858–1862.
O’Keefe, E.J., Briggaman, R.A., and Herman, B. (1987) Calciuminduced assembly of adherens junctions in keratinocytes. J. Cell
Biol., 105:807–817.
Pizarro, A., Benito, N., Navarro, P., Palacios, J., Cano, A., Quintanilla,
M., Contreras, F., and Gamallo, C. (1994) E-cadherin expression in
basal cell carcinoma. Br. J. Cancer, 69:157–162.
Plott, R.T., Amagai, M., Udey, M.C., and Stanley, J.R. (1994) Pemphigus vulgaris antigen lacks biochemical properties characteristic of
classical cadherins. J. Invest. Dermatol., 103:168–172, 1994.
Pouliot, Y. (1992) Phylogenetical analysis of the cadherin superfamily.
Bio. Essays 14:743–748.
Rimm, D.L., and Morrow, J. (1994) Molecular cloning of human
E-cadherin suggests a novel subdivision of the cadherin superfamily. Biochem. Biophys. Res. Comm., 200:1754–1761.
Ruggeri, B., Caamano, J., Slaga, T.J., Conti, C.J., Nelson, W.J., and
Klein-Szanto, A.J.P. (1992) Alterations in the expression of uvomorulin and Na1, K1-adenosine triphosphatase during mouse skin
tumor. Am. J. Pathol., 140:1179–1185.
Sano, K., Tanihara, H., Heimark, R.L., Obata, S., Davidson, M., St.
John, T., Taketani, S., and Suzuki, S. (1993) Protocadherins: a large
family of cadherin-related molecules in central nervous system.
EMBO J., 12:2249–2256.
Shimoyama, Y., Hirohashi, S., Hirano, S., Noguchi, M., Shimosato, Y.,
Takeichi, M., and Age, O. (1989) Cadherin cell-adhesion molecules
in human epithelial tissues and carcinomas. Cancer Res., 49:2128–
Shirahama, S., Furukawa, F., Wakita, H., and Takigawa, M. (1996) Eand P-cadherin expression in tumor tissues and soluble E-cadherin
levels in sera of patients with skin cancer. J. Dermatol. Sci.,
Stanley, J.R. (1995) Autoantibodies against adhesion molecules and
structures in blistering skin diseases. J. Exp. Med., 181:1–4.
Suzuki, S., Sano, K., and Tanihara, H. (1991) Diversity of the cadherin
family: evidence for eight new cadherins in nervous system. Cell
Regulation, 2:261–270.
Takeichi, M. (1977) Functional correlation between cell adhesive
properties and some cell surface protein. J. Cell Biol., 75:464–474.
Takeichi, M. (1988) The cadherins: cell-cell adhesion molecules controlling animal morphogenesis. Development, 102:639–655.
Takeichi, M. (1991) Cadherin cell adhesion receptors as a morphogenetic regulator. Science, 251:1451–1455.
Takeichi, M., Hirano, S., Matsuyoshi, N., and Fujimori, T. (1992)
Cytoplasmic control of cadherin-mediated cell-cell adhesion. Cold
Spring Harbor Symposium on Quantitative Biology, LVII: 327–334.
Takeichi, M., Matsuyoshi, N., Fujii, K., Furukawa, F., and Imamura,
S. (1990) Roles of cadherin in the development of pemphigus.
Annual report of rare and intractable disease research committee.
ed. by Imamura, S., Japanese Ministry of Health and Welfare,
Tokyo, pp. 185–187. (in Japanese).
Tang, A., Amagai, M., Granger, L.G., Stanley, J.R., and Udey, M.C.
(1993) Adhesion of epidermal Langerhans cells to keratinocytes
mediated by E-cadherin. Nature 361:82–85.
Wheelock, M.J., Buck, C.A., Bechtol, K.B., and Damsky, C.H. (1987)
Soluble 80-kd fragment of cell-CAM 120/80 disrupts cell-cell adhesion. J. Cell. Biochem., 34:187–202.
Wheelock, M.J., and Jensen, P.J. (1992) Regulation of keratinocyte
intercellular junction organization and epidermal morphogenesis by
E-cadherin. J. Cell Biol., 117:415–425.
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