The CD44v6 epitope has been widely reported to be expressed in human mammary carcinomas, yet its prognostic significance is controversial and its function in mammary tumors and mammary glands is unknown. To begin to resolve these issues, we analysed in detail the normal postnatal expression patterns and regulation of the CD44v6 epitope in murine mammary glands. We demonstrate that significant CD44v6 epitope expression is first seen during puberty, and that after puberty CD44v6 epitope expression follows the estrous cycle. CD44v6 epitope expression is observed in the myoepithelium and also less widely in luminal epithelial cells. During lactation, CD44v6 epitope expression is turned off and reappears during involution. The CD44 variant isoform bearing the v6 epitope is CD44v1-v10. Using HC11, a mammary epithelial cell line with stem cell characteristics, and facilitated by the cloning of the murine CD44 promoter, we show that growth factors and hormones which regulate ductal growth and differentiation modulate CD44 transcription. Together our data suggest that the CD44v6 epitope is expressed in mammary epithelial stems cells and in lineages derived from these cells, and that CD44v6 expression is regulated in part by hormones and growth factors such as IGF-1 and EGF which regulate the growth and differentiation of the mammary epithelium. The function of these same growth factors and hormones is often perturbed in mammary carcinomas, and we suggest that CD44v6 expression in tumors reflects this perturbation. We conclude that the expression of the CD44v6 epitope observed in some mammary tumors reflects the stem cell origin of breast tumors, and that whether or not the CD44v6 epitope is expressed in a mammary tumor is determined by the differentiation status of the tumor cells.
REFERENCES
Ball
R.
, Friis
R.
, Schoenenberger
C.
, Doppler
W.
, Groner
B.
(1988
). Prolactin regulation of-casein gene expression and of a cytosolic 120-kd protein in a cloned mouse mammary epithelial cell line.
EMBO J
7
, 2089
–2095
Bell
M.
, Cowper
A.
, Lefranc
M.
, Bell
J.
, Screaton
G.
(1998
). Influence of intron length on alternative splicing of CD44.
Mol. Cell Biol
18
, 5930
–5941
Bennett
K.
, Jackson
D.
, Simon
J.
, Tanczos
E.
, Peach
R.
, Modrell
B.
, Stamenkovic
I.
, Plowman
G.
, Aruffo
A.
(1995
). CD44 isoforms containing exon v3 are responsible for the presentation of heparin-binding growth factor.
J. Cell Biol
128
, 687
–698
Berger
M.
, Locher
G.
, Saurer
S.
, Gullick
W.
, Waterfield
M.
, Groner
B.
, Hynes
N.
(1988
). Correlation of c-erbB-2 gene amplification and protein expression in humanbreast carcinoma with nodal status and nuclear grading.
Cancer Res
48
, 1238
–1243
Chammas
R.
, Taverna
D.
, Cella
N.
, Santos
C.
, Hynes
N.
(1994
). Laminin and tenascin assembly and expression regulate HC11 mouse mammary cell differentiation.
J. Cell Sci
107
, 1031
–1040
Coleman
S.
, Daniel
C.
(1990
). Inhibition of mouse ductal morphogenesis and down-regulation of the EGF receptor by epidermal growth factor.
Dev. Biol
137
, 425
–433
Dall
P.
, Heider
K.-H.
, Sinn
H.-P.
, Skroch-Angel
P.
, Adolf
G.
, Kaufmann
M.
, Herrlich
P.
, Ponta
H.
(1995
). Comparison of immunohistochemistry and RT-PCR for detection of CD44v-expression, a new prognostic factor in human breast cancer,.
Int. J. Cancer
60
, 471
–477
Deugnier
M.-A.
, Faraldo
M.
, Rousselle
P.
, Thiery
J.-P.
, Glukhova
M.
(1999
). Cell-extracellular matrix interactions and EGF are important regulators of the basal mammary epithelial cell phenotype.
J. Cell Sci
112
, 1035
–1044
Dulbecco
R.
, Allen
W. R.
, Bologna
M.
, Bowman
M.
(1986
). Marker evolution during the development of the rat mammary gland: stem cells identified by markers and the role of myoepithelial cells.
Cancer Res
46
, 2449
–2456
Earp
H.
, Dawson
T.
, Li
X.
, Yu
H.
(1995
). Heterodimersization and functional interaction between EGF receptor family members: a new signaling paradigm with implications for breast cancer research.
Breast Cancer Res. Treat
35
, 115
–132
Foekens
J.
, Dall
P.
, Klijn
J.
, Skorch-Angel
P.
, Claasen
C.
, Look
M.
, Ponta
P.
, van Putten
W.
, Herrlich
P.
, Henzen-Logmans
S.
(1999
). Prognostic value of CD44 variant expression in primary breast cancer.
Int. J. Cancer
, 84
, 209
–215
Fox
S.
, Smith
K.
, Hollyer
J.
, Greenall
M.
, Hastrich
D.
, Harris
A.
(1994
). The epidermal growth factor receptor as a prognostic marker: results of 370 patients and review of 3009 patients.
Breast Cancer Res. Treat
29
, 41
–49
Friedrichs
K.
, Franke
F.
, Lisboa
B.-W.
, Kugler
G.
, Gille
I.
, Terpe
H.-J.
, Hölzel
F.
, Maass
H.
, Gunthert
U.
(1995
). CD44 isoforms correlate with cellular differentiation but not with prognosis is human breast cancer.
Cancer Res
55
, 5424
–5433
Hofmann
M.
, Rudy
W.
, Gunthert
U.
, Zimmer
S.
, Zawadzki
V.
, Zöller
M.
, Lichtner
R.
, Herrlich
P.
, Ponta
H.
(1993
). A link between ras and metastatic behaviour of tumor cells: ras induces CD44 promoter activity and leads to low-level expression of metastasis-specific variants of CD44 in CREF cells.
Cancer Res
53
, 1516
–1521
Hofmann
M.
, Fieber
C.
, Assmann
V.
, Göttlicher
M.
, Sleeman
J. P.
, Plug
R.
, Howells
N.
, von Stein
O.
, Ponta
H.
, Herrlich
P.
(1998
). Identification of IHABP, a 95 kDa intracellular hyaluronate binding protein.
J. Cell Sci
111
, 1673
–1684
Humphreys
R.
, Rosen
J.
(1997
). Stably transfected HC11 cells provide an in vitro and in vivo model system for studying Wnt gene function.
Cell Growth Differ
8
, 839
–849
Jansen
R.
, Joosten-Achjanie
S.
, Arends
J.
, Volovics
A.
, Hupperets
P.
, Schouten
H.
, Hillen
H.
(1998
). CD44v6 is not a prognostic factor in primary breast cancer.
Ann. Oncol
9
, 109
–111
Katagiri
Y.
, Sleeman
J.
, Fujii
H.
, Herrlich
P.
, Hotta
H.
, Tanaka
K.
, Chikuma
S.
, Yagita
H.
, Okumura
K.
, Murakami
M.
, Saiki
I.
, Chambers
A.
, Uede
T.
(1999
). CD44 variants but not CD44s cooperate with1-containing integrins to permit cells to bind to osteopontin independently of arginine-glycine-aspartic acid, thereby stimulating cell motility and chemotaxis.
Cancer Res
59
, 219
–226
Kaufmann
M.
, Heider
K.-H.
, Sinn
H.-P.
, von Minckwitz
G.
, Ponta
H.
, Herrlich
P.
(1995
). CD44 variant exon epitopes in primary breast cancer and length of survival.
Lancet
345
, 615
–619
Kleinberg
D.
(1998
). Role of IGF-1 in normal mammary development.
Breast Cancer Res. Treat
47
, 201
–208
König
H.
, Moll
J.
, Ponta
H.
, Herrlich
P.
(1996
). Trans-acting factors regulate the expression of CD44 splice variants.
EMBO J
15
, 4030
–4039
König
H.
, Ponta
H.
, Herrlich
P.
(1998
). Coupling of signal transduction to alternative pre-mRNA splicing by a composite splice regulator.
EMBO J
17
, 2904
–2913
Klijn
J.
, Berns
P.
, Schmitz
P.
, Foekens
J.
(1992
). The clinical significance of epidermal growth factor recptor (EGF-R) in human breast cancer: a review on 5232 patients.
Endocr. Rev.
13
, 3
–17
Naor
D.
, Sionov
R.
, Ish-Shalom
D.
(1997
). CD44: Structure, function and association with the malignant process.
Advan. Cancer Res
71
, 241
–319
Nordeen
S.
(1988
). Luciferase reporter gene vectors for analysis of promoters and enhancers.
BioTechniques
6
, 454
–457
Nottenburg
C.
, Rees
G.
, St John
T.
(1989
). Isolation of mouse CD44 cDNA: Structural features are distinct from the primate cDNA.
Proc. Nat. Acad. Sci. USA
86
, 8521
–8525
Omerod
E. J.
, Rudland
P.
(1986
). Regeration of mammary glands in vivo from isolated mammary ducts.
J. Embryol. Exp. Morphol
96
, 229
–243
Peyrat
J.
, Bonneterre
J.
(1992
). Type 1 IGF receptor in human breast diseases.
Breast Cancer Res. Treat
22
, 59
–67
Peyrat
J.
, Bonneterre
J.
, Hecquet
B.
, Vennin
P.
, Louchez
M.
, Fournier
C.
, Lefebvre
J.
, Demaille
A.
(1993
). Plasma insulin-like growth factor 1 (IGF-1) concentrations in human breast cancer.
Eur. J. Cancer
29
, 492
–497
Radnor
C.
(1972
). Myoepithelial cell differentiation in rat mammary glands.
J. Anat
111
, 381
–398
Reber
S.
, Matzku
S.
, Gunthert
U.
, Ponta
H.
, Herrlich
P.
, Zöller
M.
(1990
). Retardation of metastatic tumor growth after immunization with metastasis-specific monoclonal antibodies.
Int. J. Cancer
46
, 919
–927
Ruan
W.
, Catanese
V.
, Wieczorek
, Feldman
M.
, Kleinberg
D.
(1995
). Estradiol enhances the stimulatory effect of insulin-like growth factor-1 (IGF-1) on mammary development and growth hormone-induced IGF-1 messenger ribonucleic acid.
Endocrinology
136
, 1296
–1302
Rudland
P.
(1987
). Stem cells and the development of mammary cancers in experimental rats and in humans.
Cancer Metast. Rev.
6
, 55
–83
Rudy
W.
, Hofmann
M.
, Schwartz-Albiez
R.
, Zöller
M.
, Heider
K.-H.
, Ponta
H.
, Herrlich
P.
(1993
). The two major CD44 proteins expressed on a metastatic rat tumor cell line are derived from different splice variants: each one individually suffices to confer metastatic behavior.
Cancer Res
53
, 1262
–1268
Sainsbury
J.
, Farndon
J.
, Needham
G.
, Malcom
A.
, Harris
A.
(1987
). Epidermal growth factor receptor status as predictor of early recurrence of and death from breast cancer.
Lancet
1
, 1398
–1402
Schönenberger
C.-A.
, Zuk
A.
, Groner
B.
, Jones
W.
, Andres
A.-C.
) (1990
). Induction of the endogenous whey acidic protein (wap) gene and a wap-myc hybrid gene in primary murine mammary organoids.
Dev. Biol
139
, 327
–337
Seiter
S.
, Arch
R.
, Komitowski
D.
, Hofmann
M.
, Ponta
H.
, Herrlich
P.
, Matzku
S.
, Zöller
M.
(1993
). Prevention of tumor metastasis formation by anti-variant CD44.
J. Exp. Med
177
, 443
–455
Sherman
L.
, Wainwright
D.
, Ponta
H.
, Herrlich
P.
(1998
). A splice variant of CD44 expressed in the apical ectodermal ridge presents fibroblast growth factors to limb mesenchyme and is required for limb outgrowth.
Genes Dev
12
, 1058
–1071
Shtivelman
E.
, Bishop
J. M.
(1991
). Expression of CD44 is repressed in neuroblastoma cells.
Mol. Cell Biol
11
, 5446
–5453
Sinn
H.-P.
, Heider
K.-H.
, Skroch-Angel
P.
, von Minckwitz
G.
, Kaufmann
M.
, Herrlich
P.
, Ponta
P.
(1995
). Human mammary carcinomas express homologues of rat metastasis-associated variants of CD44.
Breast Cancer Res. Treat
36
, 307
–313
Sleeman
J. P.
, Arming
S.
, Moll
J.
, Hekele
A.
, Rudy
W.
, Sherman
L.
, Kreil
G.
, Ponta
H.
, Herrlich
P.
(1996
). Hyaluronate-independent metastatic behavior of CD44 variant-expressing pancreatic carcinoma cells.
Cancer Res
56
, 3134
–3141
Sonnenberg
A.
, Daams
H.
, van der Valk
M.
, Hilkens
J.
, Hilgers
J.
(1986
). Development of the mouse mammary gland: identification of stages in differentiation of luminal and myoepithelial cells using monoclonalantibodies and polyvalent antiserum against keratin.
J. Histochem. Cytochem
34
, 1037
–1046
Tempfer
C.
, Losch
A.
, Heinzl
H.
, Hausler
G.
, Hanzal
E.
, Kolbl
H.
, Breitenecker
G.
, Kainz
C.
(1996
). Prognostic value of immunohistochemically detected CD44 isoforms CD44v5, CD44v6 and CD44v7-8 in human breast cancer.
Eur. J. Cancer
32
, 2023
–2025
Tokue
Y.
, Matsumura
Y.
, Katsumata
N.
, Watanabe
T.
, Tarin
D.
, Kakizoe
T.
(1998
). CD44 variant isoform expression and breast cancer prognosis.
Jpn. J. Cancer Res
89
, 283
–290
Tölg
C.
, Hofmann
M.
, Herrlich
P.
, Ponta
H.
(1993
). Splicing choice from ten variant exons establishes CD44 variability.
Nucl. Acids Res
21
, 1225
–1229
Topper
Y.
, Freeman
C.
(1980
). Multiple hormone interactions in thedevelopmental biology of the mammary gland.
Phys. Rev.
60
, 1049
–1106
Trowbridge
I.
, Lesley
J.
, Schulte
R.
, Hyman
R.
, Trotter
J.
(1982
). Biochemical characterisation and cellular distribution of a polymorphic murine cell-surface glycoprotein expressed in lymphoid tissues.
Immunogenetics
15
, 299
–312
van Weering
D. H.
, Baas
P. D.
, Bos
J. L.
(1993
). A PCR-based method for the analysis of human CD44 splice products.
PCR Meth. Appl
3
, 100
–1066
Weiss
J. M.
, Sleeman
J. P.
, Renkl
A. C.
, Termeer
C.
, Dittmar
H.
, Taxis
S.
, Howells
N.
, Hofmann
M.
, Schöpf
E.
, Ponta
H.
, Herrlich
P.
, Simon
J. C.
(1997
). An essential role for CD44 variant isoforms in epidermal Langerhans cell and blood dendritic cell function.
J. Cell Biol
137
, 1
–11
Wellings
S.
, Jensen
H.
(1973
). On the origin and progression of ductal carcinoma in the human breast.
J. Nat. Cancer Inst
50
, 1111
–1118
Williams
J.
, Daniels
C.
(1983
). Mammary ductal elongation: differentiation of myoepithelium and basal lamina during branching morphogenesis.
Dev. Biol
97
, 274
–290
Winstanley
J.
, Cooke
T.
, Murray
G.
, Platt-Higgins.
A.
, George
W.
, Holt
S.
, Myskov
M.
, Spedding
A.
, Barraclough
B.
, Rudland
P.
(1991
). The long term prognostic significance of c-erbB-2 in primary breast cancer.
Br. J. Cancer
63
, 447
–450
Xie
W.
, Paterson
A.
, Chin
E.
, Nabell
L.
, Kudlow
J.
(1997
). Targeted expression of a dominant negative epidermal growth factor receptor in the mammary gland of transgenic mice inhibits pubertal mammary duct development.
Mol. Endocrinol
11
, 1766
–1781
Yu
Q.
, Stamenkovic
I.
(1999
). Localization of matrix metalloproteinase 9 to the cell surface provides a mechanism for CD44-mediated tumor invasion.
Genes Dev
13
, 35
–48
This content is only available via PDF.
© 2000 by Company of Biologists
2000
-PolarityCFP.png?versionId=4696)
