Recent evidence indicates that CD44, a multifunctional adhesion receptor involved in cell-cell as well as in cell-matrix interactions, plays an important role in local progression and metastasis of malignant tumors. We have studied a set of human melanoma cell lines differing in their metastatic potential in nude mice as well as in normal melanocytes for changes in CD44 expression and function. All melanocytes and melanoma cell lines tested highly expressed the CD44 standard form (CD44s, 85 kDa) but variants at low levels only. With respect to one of the CD44-associated functions primarily involved in tumor progression we found that two highly metastatic tumor cell lines, MV3 and BLM, showed fivefold higher migration rates towards hyaluronate than melanomas with low metastatic potential and normal melanocytes. Moreover, the highly metastatic cell lines expressed four- to sixfold higher levels of the CD44 epitope involved in hyaluronic acid-binding (monoclonal antibody Hermes-1) than less aggressive melanomas and melanocytes. Hermes-1 efficiently blocked haptotaxis to hyaluronate, supporting the functional relevance of this epitope. In contrast, expression levels of other CD44s epitopes recognized by seven different anti-CD44 monoclonal antibodies were unchanged, suggesting that the migratory behaviour of the cells depends on the formation of the hyaluronate-binding Hermes-1 epitope rather than on the overall CD44s surface expression, which was virtually identical in all melanoma and melanocyte cell lines tested. Differences in the accessibility of the hyaluronate-binding epitope defined by Hermes-1 correlated with the phosphorylation state of CD44s, probably reflecting different activation states of the receptor. Furthermore, immunoprecipitation and pulse/chase studies revealed a three- to fivefold increase in CD44 synthesis in the highly aggressive melanoma cells as compared to the other cell lines and the melanocytes, indicating a reduction of CD44 half-life and up-regulation of turnover. Moreover, highly aggressive melanoma cell lines were found to shed significant amounts of CD44 from the cell surface and to secrete its ligand hyaluronic acid, which may refer to an “autocrine' mechanism mediating melanoma cell motility.
REFERENCES
Aruffo
A.
, Stamenkovic
I.
, Melnick
M.
, Underhill
C. B.
, Seed
B.
(1990
). CD44 is the principal cell surface receptor for hyaluronate.
Cell
61
, 1303
–1313
Bartolazzi
A.
, Peach
R.
, Aruffo
A.
, Stamenkovic
I.
(1994
). Interaction between CD44 and hyaluronate is directly implicated in the regulation of tumor development.
J. Exp. Med
180
, 53
–66
Bazil
V.
, Horejsi
V.
(1992
). Shedding of the CD44 adhesion molecule from leukocytes induced by anti-CD44 monoclonal antibody simulating the effect of a natural receptor ligand.
J. Immunol
149
, 747
–753
Bazil
V.
, Strominger
J. L.
(1994
). Metalloprotease and serine protease are involved in cleavage of CD43, CD44 and CD16 from stimulated human granulocytes.
J. Immunol
152
, 1314
–1322
Brown
T. A.
, Bouchard
T.
, St. John
T.
, Wayner
E.
, Carter
W. G.
(1991
). Human keratinocytes express a new CD44 core protein (CD44E) as a heparan-sufate intrinsic membrane proteoglycan with additional exons.
J. Cell. Biol
113
, 207
–221
Culty
M.
, Miyake
K.
, Kincade
P. W.
, Silorski
E.
, Butcher
E. C.
, Underhill
C.
(1990
). The hyaluronate receptor is a member of the CD44 (H-CAM) family of cell surface glycoproteins.
J. Cell Biol
111
, 2765
–2774
Culty
M.
, Shizari
M.
, Thompson
E. W.
, Underhill
C. B.
(1994
). Binding and degradation of hyaluronan by human breast cancer cell lines expressing different forms of CD44: correlation with invasive potential.
J. Cell. Physiol
160
, 275
–286
Danen
E. H. J.
, van Muijen
G. N. P.
, Wiel van de Kemenade
E.
, Jansen
K. F. J.
, Ruiter
D. J.
, Figdor
C. G.
(1993
). Regulation of integrin-mediated adhesion to laminin and collagen in human melanocytes and in non-metastatic and highly metastatic human melanoma cells.
Int. J. Cancer
54
, 315
–321
Flanagan
B. F.
, Dalchau
R.
, Allen
A. K.
, Daar
A. S.
, Fabre
J. W.
(1989
). Chemical composition and tissue distribution of the human CDw44 glycoprotein.
Immunology
67
, 167
–175
Gallatin
M. W.
, Wayner
E. A.
, Hoffman
P. A.
, St. John
T.
, Butcher
E. C.
, Carter
W. G.
(1989
). Structural homology between lymphocyte receptors for high endothelium and class III extracellular matrix receptor.
Proc. Nat. Acad. Sci. USA
86
, 4654
–4658
Goebeler
M.
, Meinardus-Hager
G.
, Roth
J.
, Goerdt
S.
, Sorg
C.
(1993
). Nickel chloride and cobalt chloride, two common contact sensitizers, directly induce expression of ICAM-1, VCAM-1 and ELAM-1 by endothelial cells.
J. Invest. Dermatol
100
, 759
–765
Gunthert
U.
, Hofmann
M.
, Rudy
W.
, Reber
S.
, Zöller
M.
, Haussmann
I.
, Matzku
S.
, Wenzel
A.
, Ponta
H.
, Herrlich
P.
(1991
). A new variant of glycoprotein CD44 confers metastatic potential to rat carcinoma cells.
Cell
65
, 13
–14
Guo
Y.
, Ma
J.
, Wang
J.
, Che
X.
, Narula
J.
, Bigby
M.
, Wu
M.
, Sy
M. S.
(1994
). Inhibition of human melanoma growth and metastasis in vivo by anti-CD44 monoclonal antibody.
Cancer Res
54
, 1561
–1565
Halaban
R.
, Gosh
S.
, Duray
P.
, Kirkwood
J. M.
, Lerner
A. B.
(1986
). Human melanocytes cultured from nevi and melanomas.
J. Invest. Dermatol
87
, 95
–101
Hardwick
C.
, Hoare
K.
, Owens
R.
, Hohn
H. P.
, Hook
M.
, Moore
D.
, Cripps
V.
, Austen
L.
, Nance
D. M.
, Turley
E. A.
(1992
). Molecular cloning of a novel hyaluronan receptor that mediates tumor cell motility.
J. Cell Biol
117
, 1343
–1350
Hart
I. R.
, Birch
M.
, Marshall
J. F.
(1991
). Cell adhesion receptor expression during melanoma progression and metastasis.
Cancer Metast. Rev.
10
, 115
–128
Haynes
B. F.
, Liao
H. X.
, Patton
K. L.
(1991
). The transmembrane hyaluronate receptor (CD44): multiple functions, multiple forms.
Cancer Cell
3
, 347
–350
Herrlich
P.
, Zöller
M.
, Pals
S. T.
, Ponta
H.
(1993
). CD44 splice variants: metastases meet lymphocytes.
Immunol. Today
14
, 395
–399
Hofmann
M.
, Rudy
W.
, Zöller
M.
, Tolg
C.
, Ponta
H.
, Herrlich
P.
, Gunthert
U.
(1991
). CD44 splice variants confer metastatic behaviour in rats: homologous sequences are expressed in human tumor cell lines.
Cancer Res
51
, 5292
–5297
Jalkanen
S.
, Bargatze
R. F.
, Herron
L. R.
, Butcher
E. C.
(1986
). A lymphoid cell surface glycoprotein involved in endothelial cell recognition and lymphocyte homing in man.
Eur. J. Immunol
16
, 1195
–1202
Jalkanen
S.
, Bargatze
R. F.
, del los Toyos
J.
, Butcher
E. C.
(1987
). Lymphocyte recognition of high endothelium: antibodies to distinct epitopes of an 85–95 kD glycoprotein antigen differentially inhibit lymphocyte binding to lymph node, mucosal or synovial endothelial cells.
J. Cell Biol
105
, 983
–900
Jalkanen
S.
, Jalkanen
M.
, Bargatze
R.
, Tammi
M.
, Butcher
E. C.
(1988
). Biochemical properties of glycoproteins involved in lymphocyte recognition of high endothelial venules in man.
J. Immunol
141
, 1615
–1623
Katoh
S.
, McCarthy
J. B.
, Kincade
P. W.
(1994
). Characterization of soluble CD44 in the circulation of mice.
J. Immunol
153
, 3440
–3449
Katoh
S.
, Zheng
Z.
, Oritani
K.
, Shimozato
T.
, Kincade
P. W.
(1995
). Glycosylation of CD44 negatively regulates its recognition of hyaluronan.
J. Exp. Med
182
, 419
–429
Klein
C. E.
, Cordon-Cordo
C.
, Soehnchen
R.
, Eote
R. J.
, Oettgen
H. F.
, Eisinger
M.
, Old
L. J.
(1987
). Changes in cell surface glycoprotein expression during differentiation of human keratinocytes.
J. Invest. Dermatol
89
, 500
–506
Klein
C. E.
, Ozer
H.
, Traganos
F.
, Atzpodien
J.
, Oettgen
H.
, Old
L. J.
(1988
). A transformation-associated 130-kD cell surface glycoprotein is growth-controlled in normal human cells.
J. Exp. Med
167
, 1684
–1696
Knudson
C. B.
, Knudson
W.
(1993
). Hyaluronan-binding proteins in development, tissue homeostasis and disease.
FASEB J
7
, 1233
–1241
Lesley
J.
, Hyman
R.
, Kincade
P. W.
(1993
). CD44 and its interaction with the extracellular matrix.
Adv. Immunol
54
, 271
–335
Lesley
J.
, English
N.
, Perschl
A.
, Gregoroff
J.
, Hyman
R.
(1995
). Variant cell lines selected for alterations in the function of the hyaluronan receptor CD44 show differences in glycosylation.
J. Exp. Med
182
, 431
–437
Mackay
C. R.
, Maddox
J. F.
, Wijffels
G. L.
, Mackay
I. R.
, Walker
I. D.
(1988
). Characterization of a 95000 molecule on sheep leukocytes homologous to murine Pgp-1 and human CD44.
Immunology
65
, 93
–99
Miyake
K.
, Underhill
C. B.
, Lesley
J.
, Kincade
P. W.
(1990
). Hyaluronate can function as a cell adhesion molecule and CD44 participates in hyaluronate recognition.
J. Exp. Med
172
, 69
–75
Murakami
S.
, Shimabukuro
Y.
, Miki
Y.
, Saho
T.
, Hino
E.
, Kasai
D.
, Nozaki
T.
, Kosomoto
Y.
, Okada
H.
(1994
). Inducible binding of human lymphocytes to hyaluronate via CD44 does not require cytoskeleton association but does require new protein synthesis.
J. Immunol
152
, 467
–477
Pals
S. T.
, Hogervorst
F.
, Keizer
G. D.
, Thepen
T.
, Horst
E.
, Figdor
C. C.
(1989
). Identification of a widely distributed 90-kDa glycoprotein that is homologous to the Hermes-1 human lymphocyte homing receptor.
J. Immunol
143
, 851
–857
Peach
R. J.
, Hollenbaugh
D.
, Stamenkovic
I.
, Aruffo
A.
(1993
). Identification of hyaluronic acid binding sites in the extracellular domain of CD44.
J. Cell Biol
122
, 257
–264
Pure
E.
, Camp
R. L.
, Peritt
D.
, Panettieri
R. A.
, Lazaar
A. L.
, Nayak
S.
(1995
). Defective phosphorylation and hyaluronate binding of CD44 with point mutations in the cytoplasmic domain.
J. Exp. Med
181
, 55
–62
Screaton
G. R.
, Bell
M. V.
, Jackson
D. G.
, Cornelis
F. B.
, Gerth
U.
, Bell
J. I.
(1992
). Genomic structure of DNA encoding the lymphocyte homing receptor CD44 reveals at least 12 alternatively spliced exons.
Proc. Nat. Acad. Sci. USA
89
, 12160
–12164
Sherman
L.
, Sleeman
J.
, Herrlich
P.
, Ponta
H.
(1994
). Hyaluronate receptors: key players in growth, differentiation, migration and tumor progression.
Curr. Opin. Cell. Biol
6
, 726
–733
Stamenkovic
I.
, Amiot
M.
, Pesando
J. M.
, Seed
B.
(1989
). A lymphocyte molecule implicated in lymph node homing is a member of the cartilage link family.
Cell
56
, 1057
–1062
Stamenkovic
I.
, Aruffo
A.
, Amiot
M.
, Seed
B.
(1991
). The hematopoietic and epithelial forms of CD44 are distinct polypeptides with different adhesion potentials for hyaluronate-bearing cells.
EMBO J
10
, 343
–348
Sy
M. S.
, Guo
Y. J.
, Stamenkovic
I.
(1991
). Distinct effects of two CD44 isoforms on tumor growth in vivo.
J. Exp. Med
174
, 859
–866
Thomas
L.
, Byers
H. R.
, Vink
J.
, Stamenkovic
I.
(1992
). CD44H regulates tumor cell migration on hyaluronate-coated substrates.
J. Cell Biol
118
, 971
–977
Thomas
L.
, Etoh
T.
, Stamenkovic
I.
, Mihm
M. C.
, Byers
H. R.
(1993
). Migration of human melanoma cells on hyaluronate is related to CD44 expression.
J. Invest. Dermatol
100
, 115
–120
Tilgen
W.
, Boukamp
P.
, Breitkreutz
D.
, Dzarlieva
R. T.
, Engstner
M.
, Haag
D.
, Fusenig
N. E.
(1983
). Preservation of morphological, functional and karyotypic traits during long-term culture and in vivo passage of two human skin squamous cell carcinomas.
Cancer Res
43
, 5995
–6011
Turley
E. A.
, Tretiak
M.
(1984
). Glycosaminoglycans produced by murine melanoma variants in vivo and in vitro.
Cancer Res
45
, 5098
–5105
Turley
E. A.
, Tretiak
M.
, Tanguay
K.
(1987
). Effect of glycosaminoglycans and enzymes on the integrity of human placental amnion as a barrier to cell invasion.
J. Nat. Cancer Inst
78
, 787
–795
Turley
E. A.
(1992
). Hyaluronan and cell locomotion.
Cancer Metast. Rev.
11
, 21
–30
van Muijen
G. N. P.
, Cornelissen
L. M. H. A.
, Jansen
C. F. J.
, Ruiter
D. J.
(1989
). Progression markers in metastasizing human melanoma cells xenografted to nude mice.
Anticancer Res
9
, 879
–884
van Muijen
G. N. P.
, Jansen
C. F. J.
, Cornelissen
L. M. H. A.
, Beck
J. L. M.
, Ruiter
D. J.
(1991
). Establishment and characterization of a human melanoma cell line (MV3) which is highly metastatic in nude mice.
Int. J. Cancer
48
, 85
–91
van Muijen
G. N. P.
, Danen
E. H.
, Veerkamp
E. H.
, Ruiter
D. J.
, Lesley
J.
, van den Heuvel
L. P.
(1995
). Glycokonjugate profile and CD44 expression in human melanoma cell lines with different metastatic capacity.
Int. J. Cancer
61
, 241
–248
Versteeg
R.
, Noordermeer
I. O. A.
, Kruse-Wolters
M.
, Ruiter
D. J.
, Schrier
P. I.
(1988
). C-myc down-regulates class I HLA expression in human melanomas.
EMBO J
7
, 1023
–1029
Yang
B.
, Yang
B. L.
, Savani
R. C.
, Turley
E. A.
(1994
). Identification of a common hyaluronate binding motif in the hyaluronan binding proteins RHAMM, CD44 and link protein.
EMBO J
13
, 286
–296
Zheng
Z.
, Katoh
S.
, He
Q.
, Oritani
K.
, Miyake
K.
, Lesley
J.
, Hyman
R.
, Hamik
A.
, Parkhouse
R. M. E.
, Farr
A. G.
, Kincade
P.
(1995
). Monoclonal antibodies to CD44 and their influence on hyaluronan recognition.
J. Cell Biol
130
, 485
–495
Zwadlo
G.
, Voegeli
R.
, Schulze Osthoff
K.
, Sorg
C.
(1987
). A monoclonal antibody to a novel differentiation antigen on human macrophages associated with the down-regulatory phase of the inflammatory process.
Exp. Cell. Biol
55
, 295
–304
This content is only available via PDF.
© 1996 by Company of Biologists
1996
-PolarityCFP.png?versionId=4839)
