Previous studies have correlated release of the 92 kDa type IV collagenase/gelatinase by tumor cells in culture with metastatic potential. We have now demonstrated that the ability of tumor cells that do not express the 92 kDa gelatinase to induce release of this metalloproteinase from normal fibroblasts may also be associated with the metastatic phenotype. A transformed rat embryo cell line, 2.8, failed to release the 92 kDa gelatinase alone in culture, but gave rise to metastatic tumors whose explants contained the 92 kDa gelatinase. In contrast, a non-metastatic transformed cell line, RA3, did not express the 92 kDa gelatinase alone in culture or in tumor explants. To explore the mechanisms that might govern host-tumor cell interactions in this system, we have studied the effects of co-culture of these transformed cell lines with rat embryo fibroblasts (REF) in culture. 92 kDa gelatinase expression was induced by co-culture of 2.8 with REF, but co-culture of the non-metastatic line RA3 with REF did not result in induction of the 92 kDa gelatinase. The 92 kDa gelatinase in these co-cultures was released by the fibroblasts; methanol-fixed 2.8 cells induced 92 kDa gelatinase expression in REF, but fixed REF cells did not induce enzyme expression in 2.8 cells. This suggested that cell contact was required for induction, which was confirmed by showing that 92 kDa gelatinase induction in co-culture was abolished by separating REF from 2.8 by solute-permissive membranes. In addition, REF could not be stimulated to produce the 92 kDa gelatinase by 2.8-derived conditioned medium, by 2.8-derived extracellular matrix, or by isolated matrix components. These data indicate that metastatic tumor cells can induce 92 kDa gelatinase expression in fibroblasts through a mechanism dependent upon cell contact. In situ hybridization of nude mouse tumors derived from these transformed cell lines revealed 92 kDa gelatinase expression in the stroma of tumors from 2.8, but not in tumors from RA3. Therefore, the experiments based on in vitro co-culture of tumor cells and fibroblasts, together with the in situ localization of mRNA to host cells, suggest that host production of the 92 kDa gelatinase may occur in response to direct contact with metastatic tumor cells.

REFERENCES

Azzam
H. S.
,
Thompson
E. W.
(
1992
).
Collagen-induced activation of the Mr72,000 type IV collagenase in normal and malignant human fibroblastoid cells.
Cancer Res
52
,
4540
4544
Ballin
M.
,
Gomez
D. E.
,
Sinha
C. C.
,
Thorgeirsson
U. P.
(
1988
).
Ras oncogene mediated induction of a 92 kDa metalloproteinase; strong correlation with the malignant phenotype.
Biochem. Biophys. Res. Commun
154
,
832
838
Ballin
M.
,
Mackay
A. R.
,
Hartzler
J. L.
,
Nason
A.
,
Pelina
M. D.
,
Thorgeirsson
U. P.
(
1991
).
Ras levels and metalloproteinase activity in normal versus neoplastic rat mammary tissues.
Clin. Exp. Metastasis
9
,
179
189
Basset
P.
,
Bellocq
J. P.
,
Wolf
C.
,
Stoll
I.
,
Hutin
P.
,
Limacher
J. M.
,
Podhajcer
O. L.
,
Chenard
M. P.
,
Rio
M. C.
,
Chambon
P.
(
1990
).
A novel metalloproteinase gene specifically expressed in stromal cells of breast carcinomas.
Nature
348
,
699
704
Bauer
E. A.
,
Gordon
J. M.
,
Reddick
M.
,
Eisen
A. Z.
(
1977
).
Quantitation and immunocytochemical localization of human skin collagenase in basal cell carcinoma.
J. Invest. Dermatol
69
,
363
367
Behrendtsen
O.
,
Alexander
C. M.
,
Werb
Z.
(
1992
).
Metalloproteinases mediate extracellular matrix degradation by cells from mouse blastocyst outgrowths.
Development
114
,
447
456
Bernhard
E. J.
,
Muschel
R.
,
Hughes
E. N.
(
1990
).
Mr92, 000 gelatinase release correlates with the metastatic phenotype in transformed rat embryo cells.
Cancer Res
50
,
3872
3877
Biswas
C.
(
1985
).
Matrix influence on the tumor cell stimulation of fibroblast collagenase production.
J. Cell. Biochem
28
,
39
45
Brenner
D. A.
,
O'Hara
M.
,
Angel
P.
,
Chojkier
M.
,
Karin
M.
(
1989
).
Prolonged activation of jun and collagenase genes by tumour necrosis factor-.
Nature
337
,
661
663
Campbell
E. J.
,
Cury
J. D.
,
Shapiro
S. D.
,
Goldberg
G. I.
,
Welgus
H. G.
(
1991
).
Neutral proteinases of human mononuclear phagocytes: Cellular differentiation markedly alters cell phenotype for serine proteinases, metalloproteinases, and tissue inhibitor of metalloproteinases.
J. Immunol
146
,
1286
1293
Dooley
S.
,
Radtke
J.
,
Blin
N.
,
Unteregger
G.
(
1988
).
Rapid detection of DNA-binding factors using protein blotting and digoxigenin-UTP marked probes.
Nucl. Acids Res
16
,
11839
11945
Fini
M. E.
,
Girard
M. T.
(
1990
).
The pattern of metalloproteinase expression by corneal fibroblasts is altered by passage in cell culture.
J. Cell Sci
97
,
373
383
Girard
M. T.
,
Matsubara
M.
,
Fini
M. E.
(
1991
).
Transforming growth factor-and interleukin-1 modulate metalloproteinase expression by corneal stromal cells. Invest. Ophthalmol.
Vis. Sci
32
,
2441
2454
Gray
S. T.
,
Wilkins
R. J.
,
Yun
K.
(
1992
).
Interstitial collagenase gene expression in oral squamous cell carcinoma.
Amer. J. Pathol
141
,
301
306
Hart
I. R.
,
Goode
N. T.
,
Wilson
R. E.
(
1989
).
Molecular aspects of the metastatic cascade.
Biochim. Biophys. Acta
989
,
65
84
Herron
G. S.
,
Banda
M. J.
,
Clark
E. J.
,
Gavrilovic
J.
,
Werb
Z.
(
1986
).
Secretion of metalloproteinases by stimulated capillary endothelial cells: II. Expression of collagenase and stromelysin activities is regulated by endogenous inhibitors.
J. Biol. Chem
261
,
2814
2818
Hewitt
R. E.
,
Leach
I. H.
,
Powe
D. G.
,
Clark
I. M.
,
Cawston
T. E.
,
Turner
D. R.
(
1991
).
Distribution of collagenase and tissue inhibitor of metalloproteinases (TIMP) in colorectal tumors.
Int. J. Cancer
49
,
666
672
Huang
C.
,
Blitzer
A.
,
Abramson
M.
(
1986
).
Collagenase in human head and neck tumors and rat tumors and fibroblasts in monolayer cultures.
Ann. Otol. Rhinol. Laryngol
95
,
158
161
Huhtala
P.
,
Tuuttila
A.
,
Chow
L. T.
,
Lohi
J.
,
Keski-Oja
J.
,
Tryggvason
K.
(
1991
).
Complete structure of the human gene for 92-kDa type IV collagenase. Divergent regulation of expresssion for the 92-and 72-kilodalton enzyme genes in HT-1080 cells.
J. Biol. Chem
266
,
16485
16490
Hulkower
K. I.
,
Sagi-Eisenberg
R.
,
Traub
L. M.
,
Georgescu
H. I.
,
Evans
C. H.
(
1991
).
Interleukin-1 and protein kinase C: identification of a novel, 35 kDa cytosolic substrate.
Agents Actions
34
,
278
281
Karelina
T. V.
,
Hruza
G. J.
,
Goldberg
G. I.
,
Eisen
A. Z.
(
1993
).
Localization of 92-kDa type IV collagenase in human skin tumors: comparison with normal human fetal and adult skin.
J. Invest. Derm
100
,
159
165
Librach
C. L.
,
Werb
Z.
,
Fitzgerald
M. L.
,
Chiu
K.
,
Corwin
N. M.
,
Esteves
R. A.
,
Grobelny
D.
,
Galardy
R.
,
Damsky
C. H.
(
1991
).
92-kD type IV collagenase mediates invasion of human cytotrophoblasts.
J. Cell. Biol
113
,
437
449
Liotta
L. A.
,
Stetler-Stevenson
W. G.
(
1991
).
Tumor invasion and metastasis: an imbalance of positive and negative regulation.
Cancer Res
51
,
5054
–.
Matrisian
L. M.
(
1990
).
Metalloproteinases and their inhibitors in matrix remodeling.
Trends Genet
6
,
121
125
Moll
Y. M.
,
Youngleib
G. L.
,
Rosinski
K. B.
,
Quigley
J. P.
(
1990
).
Tumor promoter-stimulated Mr92, 000 gelatinase secreted by normal and malignant human cells: isolation and characterization of the enzyme from HT1080 tumor cells.
Cancer Res
50
,
6162
6170
Nabeshima
K.
,
Lane
W. S.
,
Biswas
C.
(
1991
).
Partial sequencing and characterization of the tumor cell-derived collagenase stimulatory factor.
Arch. Biochem. Biophys
285
,
90
96
Nishizuka
Y.
(
1984
).
The role of protein kinase C in cell surface signal transduction and tumor promotion.
Nature
308
,
693
698
Okada
Y.
,
Gonoji
Y.
,
Naka
K.
,
Tomita
K.
,
Nakanishi
I.
,
Iwata
K.
,
Yamashita
K.
,
Hayakawa
T.
(
1992
).
Matrix metalloproteinase 9 (92-kDa gelatinase/type IV collagenase) from HT 1080 human fibrosarcoma cells. Purification and activation of the precursor and enzymic properties.
J. Biol. Chem
267
,
21712
21719
Polette
M.
,
Clavel
C.
,
Muller
D.
,
Abecassis
J.
,
Binninger
I.
,
Birembaut
P.
(
1991
).
Detection of mRNAs encoding collagenase I and stromelysin 2 in carcinomas of the head and neck by in situ hybridization.
Inv. Metast
11
,
76
83
Poulsom
R.
,
Pignatelli
M.
,
Stetler-Stevenson
W. G.
,
Liotta
L. A.
,
Wright
P. A.
,
Jeffery
R. E.
,
Longcraft.
J. M.
,
Rogers
L.
,
Stamp
G. W. H.
(
1992
).
Stromal expression of 72 kda type IV collagenase (MMP-2) and TIMP-2 mRNAs in colorectal neoplasia.
Amer. J. Pathol
141
,
389
396
Prescott
J.
,
Troccoli
N.
,
Biswas
C.
(
1989
).
Coordinate increase in collagenase mRNA and enzyme levels in human fibroblasts treated with the tumor cell factor, TCSF.
Biochem. Int
19
,
257
266
Pyke
C.
,
Ralfkiaer
E.
,
Huhtala
P.
,
Hurskainen
T.
,
Dano
K.
,
Tryggvason
K.
(
1992
).
Localization of messenger RNA for Mr72, 000 and 92, 000 type IV collagenases in human skin cancer by in situ hybridization.
Cancer Res
52
,
1336
1341
Pyke
C.
,
Ralfkiaer
E.
,
Tryggvason
K.
,
Dano
K.
(
1993
).
Messenger RNA for two type IV collagenases is located in stromal cells in human colon cancer.
Amer. J. Pathol
142
,
359
365
Salo
T.
,
Lyons
J. G.
,
Rahemtulla
F.
,
Birkedal-Hansen
H.
,
Larjava
H.
(
1991
).
Transforming growth factor-1 up-regulates type IV collagenase expression in cultured human keratinocytes.
J. Biol. Chem
266
,
11436
11441
Sato
H.
,
Seiki
M.
(
1993
).
Regulatory mechanism of 92 kDa type IV collagenase gene expression which is associated with invasiveness of tumor cells.
Oncogene
8
,
395
405
Teale
D. M.
,
Khidair
I. A.
,
Potter
C. W.
,
Rees
R. C.
(
1988
).
Modulation of type IV collagenase and plasminogen activator in a hamster fibrosarcoma by basement membrane components and lung fibroblasts.
Br. J. Cancer
57
,
475
480
Turpeenniemi-Hujanen
T.
,
Thorgeirsson
U. P.
,
Rao
C. N.
,
Liotta
L. A.
(
1986
).
Laminin increases the release of type IV collagenase from malignant cells.
J. Biol. Chem
261
,
1883
1889
Unemori
E. N.
,
Hibbs
M. S.
,
Amento
E. P.
(
1991
).
Constitutive expression of a 92-kD gelatinase (type IV collagenase) by rheumatoidsynovial fibroblasts and its induction in normal human fibroblasts by inflammatory cytokines.
J. Clin. Invest
88
,
1656
1662
Urbanski
S. J.
,
Edwards
D. R.
,
Maitland
A.
,
Leco
K. J.
,
Watson
A.
,
Kossakowska
A. E.
(
1992
).
Expression of metalloproteinases and their inhibitors in primary pulmonary carcinomas.
Br. J. Cancer
66
,
1188
1194
Watanabe
H.
,
Nakanishi
I.
,
Yamashita
K.
,
Hayakawa
T.
,
Okada
Y.
(
1993
).
Matrix metalloproteinase-9 (92 kDa gelatinase/type IV collagenase) from U937 monoblastoid cells: correlation with cellular invasion.
J. Cell Sci
104
,
991
999
Welgus
H. G.
,
Campbell
E. J.
,
Cury
J. D.
,
Eisen
A. Z.
,
Senior
R. M.
,
Wilhelm
S. M.
,
Goldberg
G. I.
(
1990
).
Neutral metalloproteinases produced by human mononuclear phagocytes: enzyme profile, regulation and expression during cellular development.
J. Clin. Invest
86
,
1496
1502
Wilhelm
S. M.
,
Collier
I. E.
,
Marmer
B. L.
,
Eisen
A. Z.
,
Grant
G. A.
,
Goldberg
G. I.
(
1989
).
SV-40-transformed human lung fibroblasts secrete a 92-kDa type IV collagenase which is identical to that secreted by normal human macrophages.
J. Biol. Chem
264
,
17213
17221
Woolley
D. E.
,
Grafton
C. A.
(
1980
).
Collagenase immunolocalization studies of cutaneous secondary melanomas.
Br. J. Cancer
42
,
260
265
Yamagata
S.
,
Ito
Y.
,
Tanaka
R.
,
Shimuzu
S.
(
1988
).
Gelatinases of metastatic cell lines of murine colon carcinoma as detected by substrate-gel electrophoresis.
Biochem. Biophys. Res. Commun
151
,
158
162
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