Content uploaded by Selman Sokmen
Author content
All content in this area was uploaded by Selman Sokmen on Apr 19, 2020
Content may be subject to copyright.
www.elsevier.de/
p
r
p
Pathology – Research and Practice 200 (2004) 379–387
ORGINAL ARTICLE
Prognostic significance of matrix metalloproteinase-2, cathepsin D,
and tenascin-C expression in colorectal carcinoma
$
Banu Sis
a,
*,
.
Ozgul Sa$
gol
a
, Ali K.upelio$
glu
a
, Selman Sokmen
b
, Cem Terzi
b
,
Mehmet Fuzun
b
, Erdener
.
Ozer
a
, Paul Bishop
c
a
Department of Pathology, School of Medicine, Dokuz Eyl .
ul University, Inciraltı, Izmir 35340, Turkey
b
Department of Surgery, School of Medicine, Dokuz Eyl .
ul University, Izmir, Turkey
c
Wythenshave Hospital, South Manchester, UK
Received 28 December 2002; accepted 20 February 2004
Abstract
Matrix metalloproteinase-2 (MMP-2) and cathepsin D (CD) play a significant role in degrading the components of
basement membrane and extracellular matrix (ECM), whereas tenascin-C (TN-C) is a glycoprotein of the ECM related
to cell adhesion and detachment. These proteins have been implicated in tumor invasion and metastasis. Therefore, we
aimed at investigating the prognostic significance of MMP-2, CD, and TN-C expressions in primary colorectal cancer.
Overall, 112 colorectal adenocarcinomas were included in the present study. MMP-2, CD, and TN-C expressions
were evaluated by immunohistochemistry and correlated with clinicopathologic prognostic parameters and survival.
Diffuse stromal TN-C immunostaining was found to be significantly correlated with advanced stage and shorter
survival time (p¼0:002 and 0.02, respectively). MMP-2 expression was found to correlate with lymph vessel invasion
(p¼0:006) and stage (p¼0:03). CD expression was related to depth of invasion (p¼0:005). No significant
relationship was found between survival and MMP-2 and CD expression (p>0:05). In multivariate analysis, stage and
vascular invasion were independent prognostic factors, whereas TN-C did not retain a clear independent relationship
to survival (p>0:05).
Our findings suggest that TN-C expression may be a potential prognostic marker in colorectal carcinoma. However,
MMP-2 and CD do not appear to be significant indicators of survival.
r2004 Elsevier GmbH. All rights reserved.
Keywords: Matrix metalloproteinase-2; Cathepsin-D; Tenascin-C; Colorectal carcinoma; Prognosis
Introduction
Degradation of basement membrane (BM) and
extracellular matrix (ECM) plays a crucial role in tumor
invasion and metastasis. Invasion of the matrix requires
active enzymatic degradation of the ECM components.
Tumor cells secrete proteolytic enzymes themselves or
induce host cells to secrete proteases [7].ECMdegrada-
tion by proteases is involved not only in local invasion,
but also in several stages of metastatic cascade, including
angiogenesis, intravasation, and extravasation [12].
The proteases involved in ECM degradation in tumor
invasion and metastasis are subdivided into four
classes: serine, cysteine, aspartic, and matrix metallo-
proteinases (MMPs). Of those, MMP-2 is responsible
ARTICLE IN PRESS
$
This study was presented in the International Conference on
Invasion and Metastasis, 14–17 June 2001, Athens, Greece.
*Corresponding author. Fax: +90-232-2777274.
E-mail address: banu.sis@deu.edu.tr (B. Sis).
0344-0338/$ - see front matter r2004 Elsevier GmbH. All rights reserved.
doi:10.1016/j.prp.2004.02.012
ARTICLE IN PRESS
for degradation of type IV collagen, which is the major
structural protein in BM [3,4,12,27,35]. Therefore,
activation of MMP-2 is a crucial step triggering the
cascade of tumor invasion and metastasis. The prog-
nostic significance of MMP-2 overexpression has been
shown in several human neoplasias, such as breast,
ovarian, head and neck, and hepatocellular carcinomas
[20,27,36]. However, there is limited evidence that
MMP-2 overexpression is associated with advanced
stage and higher metastatic potential in colorectal
carcinoma [16,29].
Cathepsin D (CD) plays a proteolytic role in the
digestion of ECM components, and has been implicated
in tumor invasion and metastasis. Increased CD
expression has been related to poor prognosis in a
variety of malignancies, particularly in breast carcinoma
[14]. CD overexpression has also been shown in colonic
disease, including inflammatory bowel diseases, color-
ectal adenomas, and carcinomas. However, the prog-
nostic value of CD overexpression in colorectal
carcinomas is still unclear.
Alterations in ECM composition occur during tumor
progression [22]. Tenascin-C (TN-C), a glycoprotein of
the ECM, is involved in embryogenesis, wound healing,
involution and neoplastic growth related to cell adhe-
sion, migration, cell growth, and angiogenesis [21,22].
TN-C overexpression has been reported in a variety of
malignant tumors, including breast, larynx, lung,
pancreas, gastric cancer, and malignant melanoma
[9,22,24]. The data related to TN-C expression in
colorectal carcinomas are limited and controversial,
and its prognostic significance remains to be determined
[15,18,24,31].
In this study, immunohistochemical methods were
used to investigate the expression of MMP-2, CD, and
TN-C in primary colorectal carcinoma and its relation-
ship with clinicopathologic parameters and survival.
Materials and methods
Tissue sections were cut from the archival paraffin-
embedded colectomy specimens obtained from 112
patients with primary colorectal adenocarcinomas diag-
nosed between 1991 and 1997. Clinical data were
obtained from patients’ records, and hematoxylin–eosin
sections of the specimens were reviewed by one
investigator (B.S.). Conventional histopathologic prog-
nostic parameters in colorectal carcinoma, such as
tumor grade, depth of invasion, lymph node involve-
ment, and vascular and perineural invasion, were noted.
The tumors were staged according to the modified
Astler–Coller staging system [5] and graded according to
the World Health Organization criteria [6]. From each
tumor, the most representative paraffin block was
chosen, and 5 mm thick sections were taken onto poly
l-lysin-coated slides for immunostaining.
Immunohistochemistry
The streptavidin–biotin–immunoperoxidase method
was applied using primary monoclonal antibodies
against MMP-2 (prediluted, Neomarkers, USA), CD
(1:300, Dako Corp., Denmark) and TN-C (1:25, Dako
Corp., Denmark). Briefly, the sections were deparaffi-
nized and rehydrated, immersed in distillated water, and
endogenous peroxidase activity was blocked using a
0.3% solution of hydrogen peroxidase in phosphate-
buffered saline (0.01 mol/l, pH 7.5) at room temperature
for 10 min. For TN-C staining, the sections were treated
with trypsin in phosphate-buffered solution for 60 min
at 50C for epitope retrieval. Primary antibodies were
applied for 60 min at room temperature and washed in
Tris buffer. Biotinylated secondary antibodies and
streptavidin-peroxidase complex (Dako Corp., Copen-
hagen, Denmark) were added consecutively for 10 min
at room temperature and washed in Tris buffer.
Peroxidase activity was visualized with 0.03% 3-30
diaminobenzidine tetrahydrochloride (Sigma Chemical
Co., St. Louis, MO, USA) applied for 5 min. After
rinsing in deionized water and counterstaining in
hematoxylin, the slides were dehydrated and mounted.
Appropriate tissue sections, serving as positive and
negative controls for each primary antibody, were also
labeled.
Evaluation of staining
CD immunoreactivity was observed in both tumor
cells and stromal cells. Cytoplasmic CD staining in
tumor cells was semiquantitatively scored as negative
(score 0), less than 25% of the cells as positive (score 1),
25–50% of the cells as positive (score 2), greater than
50% of the cells as positive (score 3) [26]. Immunostain-
ing of stromal cells was considered to be positive if more
than 15% of cells were stained [40]. We also classified
the staining patterns in tumor cells as supranuclear,
basal, and diffuse cytoplasmic [1].
MMP-2 immunoreactivity in the cytoplasm of tumor
cells was scored as follows: no staining (score 0), less
than 10% of the cells positive (score 1), 10–40% of the
cells positive (score 2), greater than 40% of the cells
positive (score 3) [23]. Stromal cells were immunostained
and subdivided into two groups: positive and negative
[16].
TN-C positivity in tumor was scored according to the
extent of immunostaining in tumor stroma: 0–25%
positivity (score 1), 25–50% positivity (score 2), 50–75%
positivity (score 3), 75–100% positivity (score 4) [18].
Staining patterns were also evaluated as subglandular
B. Sis et al. / Pathology – Research and Practice 200 (2004) 379–387380
ARTICLE IN PRESS
(outlining the border of the neoplastic glands) and
diffuse stromal pattern [24]. In addition, TN-C cyto-
plasmic staining in tumor cells was scored as follows: no
staining (score 0), 1–25% of the cells positive (score 1),
25–75% of the cells positive (score 2), greater than 75%
of the cells positive (score 3). Statistical analysis was
performed using a computer software (SPSS 5.0, Chicago,
IL, USA). The probability level of 0.05 or less was chosen
to represent statistical significance. The relationship
between immunohistochemical scores and the clinico-
pathological parameters was evaluated by the chi-square
test, Spearman’s correlation test, and Kendall’s Tau-B
correlation test. Survival analysis was carried out using
Cox regression test and the Kaplan Meier method.
Results
The mean age of patients was 61.2713.6 years (range
24–95). There were 46 (41.1%) women and 66 (58.9%)
men. Median follow-up period was 66 months (range
48–120 months). During the follow-up period, 38
(33.9%) patients died: 37 from distant metastasis and
one from postoperative complications. Median survival
time was 42.5 months (range 1–108). Twelve patients
were lost for follow-up.
According to the modified Astler–Coller staging
system, 11 (9.8%) cases were stage B1, 52 (46.4%) were
stage B2, 3 (2.7%) were stage C1, 36 (32.1%) were stage
C2, and 10 (8.9%) were stage D. Lymph node metastasis
was found in 47 (42%) cases, whereas the remaining
ones were node-negative. Distant metastasis was found
in 10 (8.9%) cases. Twenty cases (18%) were well
differentiated, 63 (56%) were moderately, and 29 (26%)
were poorly differentiated.
CD immunohistochemistry
CD staining was observed in both neoplastic and
stromal cells as brown, fine to coarse granular cyto-
plasmic staining. CD immunostaining was positive in
stromal cells in 102 (91.1%) of 112 cases (Fig. 1). The
positive stromal cells were mainly fibrohistiocytic cells
focusing on the vicinity of the tumor cells.
Tumor cells were stained with CD antibody in 109
(97.3%) out of 112 cases. Score 1 was observed in 10
(8.9%) cases, score 2 in 29 (25.9%) cases, and score 3 in
70 (62.5%) cases. Immunostaining pattern was supra-
nuclear in 17 (15.2%) cases, basal in 10 (8.9%) cases,
and diffuse cytoplasmic in 82 (73.2%) cases. Although
there was a statistically significant relationship in terms
of depth of invasion between cases with and without CD
expression in stromal cells (p¼0:005), we found no
positive or negative correlation between these para-
meters. A positive correlation was found between CD
expression in tumor cells and patient age (p¼0:04;
Fig. 1. CD immunostaining in stromal cells of colorectal adenocarcinoma (immunoperoxidase, 100 ).
B. Sis et al. / Pathology – Research and Practice 200 (2004) 379–387 381
ARTICLE IN PRESS
r¼0:19). In addition, the CD immunostaining pattern
in tumor cells showed a significant relationship with
both tumor grades and depths of invasion (p¼0:007
and 0.03, respectively). However, a correlation test did
not reveal a direct relationship. No significant relation-
ship was found between CD expression and survival,
stage, and lymph node metastasis (p>0:05).
MMP-2 immunohistochemistry
MMP-2 immunoreactivity presented as diffuse cyto-
plasmic staining in both tumor cells (Fig. 2) and stromal
cells. MMP-2 immunoreactivity in stromal cells was
positive in 30 (26.8%) out of 112 cases. Immunoreactive
stromal cells were mostly fibrohistiocytic cells near the
neoplastic glands. MMP-2 immunostaining in tumor
cells was observed in 46 (41.1%) cases (score 1: 25 cases
(22.3%), score 2: 13 cases (11.6%), score 3: 8 cases
(7.1%)). MMP-2 expression in tumor cells was found to
be directly correlated with lymph vessel invasion
(p¼0:006;r¼0:178). A significant relationship was
also found between MMP-2 expression and stage in
both tumor cells and stromal cells (p¼0:03 and 0.01,
respectively), but there was no correlation between
them. No significant relationship was found between
MMP-2 expression and the other clinicopathological
parameters and survival.
TN-C immunohistochemistry
TN-C expression was determined mainly in tumor
stroma (Fig. 3). Some of the tumor cells also showed
cytoplasmic TN-C immunoreactivity. Ninety-seven
cases (95.5%) showed stromal TN-C expression (score
1: 16 cases (14.3%), score 2: 23 cases (20.5%), score 3:
32 cases (28.6%), score 4 :36 cases (32.1%)). Sixty-nine
(61.6%) and 38 (33.9%) cases showed diffuse and
subglandular stromal TN-C immunostaining, respec-
tively. In 20 (17.9%) cases, tumor-cell cytoplasms also
expressed TN-C (score 1: 5 cases (4.5%), score 2: 6 cases
(5.4%), score 3: 9 cases (8.0%)). Diffuse stromal TN-C
expression was significantly related to advanced stage
and to the presence of lymph node metastasis (p¼0:002
and 0.001, r¼0:275;0.291, respectively). In addi-
tion, strong (p¼0:04) and diffuse (p¼0:02) stromal
TN-C expression was significantly related to shorter
survival time (Figs. 4 and 5).
In multivariate analysis, tumor stage (pp0:001) and
venous invasion (p¼0:007) were independent prognos-
tic factors, whereas stromal TN-C did not retain a clear
independent relationship to survival (p>0:05). No
significant relationship was found between TN-C
expression in tumor cells and other clinicopathological
parameters (p>0:05).
A positive correlation was found between MMP-2
expression in stromal cells and stromal TN-C expression
(p¼0:004;r¼0:25). CD expression in tumor cells and
Fig. 2. Cytoplasmic staining of MMP-2 in tumor cells (immunoperoxidase, 200 ).
B. Sis et al. / Pathology – Research and Practice 200 (2004) 379–387382
ARTICLE IN PRESS
Fig. 3. Diffuse stromal TN-C expression in colorectal adenocarcinoma (immunoperoxidase, 200 ).
MONTHS
120100806040200
SURVIVAL
1.0
0.0
Stromal tenascin
c
++++
+++
++
+
Fig. 4. The relation of stromal tenascin-C expression to survival (Kaplan Meier).
B. Sis et al. / Pathology – Research and Practice 200 (2004) 379–387 383
ARTICLE IN PRESS
MMP-2 expression in tumor cells also showed a
correlation (p¼0:03;r¼0:18).
Discussion
The major important prognostic factors in colorectal
carcinoma are the extent of intestinal wall invasion,
lymph node metastasis, and consequently tumor stage
[13,28]. Nevertheless, 5-year overall survival can show
significant variations under the same stage groupings,
indicating the role of other prognostic variables.
Recently, tumor biological factors and the molecular
components of tumor-associated matrix have been
investigated as additional prognostic factors in color-
ectal carcinomas.
MMP-2, a 72 kDA gelatinase, is responsible for the
digestion of type IV collagen, and is considered to play a
role in the invasion of malignant cells [3,4,27]. MMP-2
expression has been shown in several human malig-
nancies. MMP-2 has been reported to be expressed
mostly in stromal cells surrounding tumor cells and also
by the tumor cells themselves. The stromal cells
producing MMP-2 are mainly fibrohistiocytic or en-
dothelial cells [4,37]. It has been suggested that tumor
cells secrete some paracrine substances that stimulate
surrounding stromal cells to produce proteinases [4].A
recently identified factor, EMMPRIN, has been shown
to induce MMP-2 expression by fibroblasts [32]. In the
present study, both of these two localizations of MMP-2
immunoreactivity were observed. In addition, the
immunostaining pattern was observed as diffuse cyto-
plasmic staining. Different intracellular localizations of
MMP-2 positivity, e.g. paranuclear or membranous,
have also been reported [36,39]. Talvensaari-Mattila
et al. [36] demonstrated MMP-2 immunoreactivity in the
periphery of the cells in cervical intraepithelial neopla-
sia, but it was diffuse in the cytoplasm of the invasive
carcinoma cells. These observations suggest that
changes in intracellular localizations of MMP-2 expres-
sion can be associated with malignant transformation of
preneoplastic lesions.
The findings regarding the effect of MMP-2 expres-
sion on tumor behavior in different organ systems are
controversial. Kusukawa et al. [25] suggested that
MMP-2 expression in oral cavity carcinomas was
correlated with lymph node metastasis. By contrast,
Charous et al. [4] did not find such a correlation in head
and neck carcinomas. In a recent study, we have
demonstrated MMP-2 expression in laryngeal preneo-
plastic and neoplastic lesions. In that study, MMP-2
MONTHS
120100806040200
SURVIVAL
1.0
.9
.8
.7
.6
.5
.4
.3
.2
.1
0.0
Tenascin C pattern
Diffuse
Subglandular
Fig. 5. The relation of tenascin-C pattern to survival (Kaplan Meier).
B. Sis et al. / Pathology – Research and Practice 200 (2004) 379–387384
ARTICLE IN PRESS
expression has been observed in 100% of patients with
laryngeal carcinoma, but no significant relationship with
stage, lymph node metastasis, or differentiation has been
found [33].
Different investigators reported increased MMP-2
expression in colorectal carcinomas compared to ade-
nomas and healthy mucosa [8,12,29,39]. D’Errico et al.
[8] found increased MMP-2 expression in carcinomas
with Dukes’ stage C to A/B. In the present study, we
noted a significant relationship between stage and
MMP-2 expression in both tumor and stromal cells,
although a correlation did not exist. Moreover, MMP-2
expression in tumor cells was closely related to lymph
vessel invasion, supporting the influence of MMP-2
expression on invasion of cancer cells. Conversely, we
found no statistically significant relationship with tumor
differentiation or survival.
CD is considered to play a significant proteolytic role
in tumor invasion and metastasis. Besides its digestive
role, it has mitogenic properties and induces angiogen-
esis by stimulating the release of basic fibroblast growth
factor [14]. In breast cancer, CD overexpression has
been found to be associated with poor overall survival
[10]. In previous reports, it has been found to be a
marker of poor prognosis in node-negative breast cancer
[41]. However, the following studies demonstrated CD
as an independent marker of poor prognosis in all
groups of breast cancer patients [34,41].
CD expression has been shown in non-neoplastic and
neoplastic diseases of the colon. Galandiuk et al. [11]
reported on higher levels of CD in colon carcinomas
compared to inflammatory bowel diseases and FAP.
Arao et al. [1] suggested that the pattern of CD
immunostaining may change during the adenoma–
carcinoma sequence. The authors found supranuclear
staining in all adenomas and in some cases of
carcinomas, whereas basal or diffuse cytoplasmic stain-
ing was noted only in carcinomas. In the present study,
we observed supranuclear, basal, and diffuse cytoplas-
mic CD staining in 15.2%, 8.9%, and 73.2% of
colorectal cacinomas, respectively. These findings ap-
pear to reflect the loss of cellular polarization in cancer
cells as suggested by Arao et al.
The data regarding the prognostic significance of CD
expression in colorectal carcinomas are controversial.
Mayer et al. [26] determined increased CD immunos-
taining intensity in Dukes’ stage C and D tumors, but
found no relationship with survival. By contrast, Iochim
et al. [17] did not find any correlation between CD
expression and stage or tumor grade. Theodoropoulos
et al. [38] correlated stromal-cell-associated CD expres-
sion with poor prognosis and suggested that CD
expression in stromal cells might be related to a more
invasive phenotype in colorectal carcinoma. However,
Caruso et al. [2] found no relationship between CD
expression by stromal cells and the clinicopathologic
parameters or survival. In the present study, we
observed CD expression in both tumor and stromal
cells. Although we determined an association between
CD expression by stromal cells and the depth of
invasion, we did not find any relationship with stage,
metastasis, or overall survival. We thus suggest that
stromal expression of CD in colorectal carcinomas may
have a limited role in tumor progression, although it
does not appear to predict survival.
TN-C is a hexameric ECM glycoprotein that is
upregulated during neoplastic growth. It has multiple
cell binding domains that mediate adhesion and migra-
tion, and also induces cell proliferation possibly owing
to epidermal growth factor-like domains [19,22].TN-C
expression has been reported to occur in several tumors
[9,22,42]. Juhasz et al. [22] correlated TN-C upregula-
tion with metastasis, early recurrence, and lethal out-
come in laryngeal and hypopharyngeal cancers. Jahkola
et al. [19] associated TN-C expression with a high
proliferative rate, and proposed TN-C as a useful
marker regarding the prediction of both local and
distant recurrence in early breast cancer. However, Pilch
et al. [30] associated TN-C expression with a better
prognosis in cervical cancer, and concluded that TN-C is
an indicator of an adequate biological defense.
Riedl et al. [31] determined a distinct increase in the
TN-C content in colon carcinomas in contrast to normal
levels in adenomas, and suggested an association
between TN-C expression and malignant progression.
Hanamura et al. [15] reported on a positive relationship
between the degree of mRNA expression of TN-C and
the depth of invasion and lymph node metastasis. In
that study, the signals were detected mainly in myofi-
broblasts in tumor stroma and also in cancer cells in one
case of the 29 colon cancers. Iskaros et al. [18] observed
expression of TN-C in well-differentiated colon cancers
as thick fibers surrounding neoplastic glands, whereas in
poorly differentiated cancers, the fibers were distributed
in an interstitial pattern. In that study, they related the
overexpression of TN-C to better survival. Similarly,
Kressner et al. [24] found two different staining patterns
of TN-C expression in tumor stroma, such as diffuse
stromal fibrillar staining and subglandular staining. In a
univariate analysis, however, they determined a sig-
nificant association between diffuse stromal staining and
shorter survival.
In the present study, TN-C expression occurred in
two different staining patterns, a diffuse stromal pattern
and a subglandular pattern, which is consistent with the
observations of Kressner et al. [24]. Diffuse stromal TN-
C expression has been found to be significantly related
to advanced stage and lymph node metastasis. More-
over, in a univariate analysis, patients with strong and
diffuse stromal TN-C expression were found to have a
shorter survival than those with weak and subglandular
stromal TN-C expression.
B. Sis et al. / Pathology – Research and Practice 200 (2004) 379–387 385
ARTICLE IN PRESS
In addition to the stromal expression of TN-C, we
also observed TN-C expression in some of the cancer
cells. In a recent study, Yoshida et al. [42] observed
TN-C immunoreactivity in tumor cells adjacent to the
stroma in larynx cancer, and suggested that TN-C
secreted by tumor cells might be involved in autocrine
proliferation and migration of cancer. In the present
study, TN-C positivity in tumor cells was diffusely
distributed in complex neoplastic glands, not only in
cells adjacent to the stroma.
In addition, there was a positive correlation between
stromal-cell-associated MMP-2 expression and stromal
TN-C expression, supporting the theory that MMPs are
induced by the production of TN-C [21]. We also found
a correlation between tumor-cell-associated MMP-2 and
CD expression, reflecting the complex proteolytic
activity of cancer cells enhancing the invasion potential.
In a previous study, Tetu et al. [37] also found an
association between MMP-2 and CD expression, which
is consistent with our findings.
To sum up, in the present study, overexpression of
TN-C has been found to be closely related to
shorter survival. These findings suggest that TN-C
may be a potential new prognostic marker in colorectal
carcinoma.
References
[1] J. Arao, H. Fukui, Y. Ono, et al., Immunohistochemical
localization of cathepsin D in colorectal tumors, Dis.
Colon Rectum 43 (2000) 396–401.
[2] M.L. Caruso, A.M. Valentini, Immunohistochemical p53
overexpression correlated to c-erbB-2 and cathepsin D
proteins in colorectal cancer, Anticancer Res. 16 (1996)
3813–3818.
[3] A.F. Chambers, L.M. Matrisian, Changing views of the
role of matrix metalloproteinases in metastasis, J. Natl.
Cancer Inst. 89 (1997) 1260–1270.
[4] S.J. Charous, G.P. Stricklin, J.L. Netterville, et al.,
Expression of matrix metalloproteinases in head and
neck squamous cell carcinoma, Ann. Otol. Rhinol.
Laryngol. 106 (1997) 271–278.
[5] A.M. Cohen, D. Kelsen, L. Saltz, et al., Adjuvant therapy
for colorectal cancer, Curr. Problems Surgery 34 (1997)
611–612.
[6] H.S. Cooper, Diagnostic Surgical Pathology, 3rd Edition,
Lippincott Williams & Wilkins, Philadelphia, 1999, pp.
1413–1468.
[7] R.S. Cotran, V. Kumar, S. Robbins, Pathologic Basis of
Disease, 5th Edition, W.B.Saunders Company, Philadel-
phia, 1994, pp. 755–831.
[8] A. D’Errico, S. Garbisa, L.A. Liotta, et al., Augmenta-
tion of type IV collagenase, laminin receptor, and Ki-67
proliferation antigen associated with human colon, gastric
and breast carcinoma progression, Mod. Pathol. 4 (1991)
239–246.
[9] K. Emoto, Y. Yamada, H. Sawada, et al., Annexin II
overexpression correlates with stromal Tenascin-C over-
expression. A prognostic marker in colorectal carcinoma,
Cancer 92 (2001) 1419–1426.
[10] J.A. Foekens, M.P. Look, J.B. Vries, et al., Cathepsin
D in primary breast cancer: prognostic evaluation
involving 2810 patients, Br. J. Cancer 79 (2) (1999)
300–307.
[11] S. Galandiuk, S. Miseljic, A. Yang, et al., Expression of
hormone receptors, cathepsin D, and HER-2/neu onco-
protein in normal colon and colonic disease, Arch. Surg.
128 (1993) 637–642.
[12] E.A. Garbett, M.W.R. Reed, N.J. Brown, Proteolysis in
human breast and colorectal cancer, Br. J. Cancer 81
(1999) 287–293.
[13] L. Gennari, R. Doci, C. Rossetti, Prognostic factors in
colorectal cancer, Hepato-Gastroenterology 47 (2000)
310–314.
[14] A. Goussia, E. Ioachim, D. Peschos, et al., Immunohis-
tochemical expression of cathepsin D in laryngeal
epithelial lesions: correlation with CD44 expression, p53
and Rb status and proliferation associated indices,
Anticancer Res. 19 (1999) 3055–3060.
[15] N. Hanamura, T. Yoshida, E. Matsumoto, Y. Kawarada,
T. Sakakura, Expression of fibronectin and tenascin-C
mRNA by myofibroblasts, vascular cells and epithelial
cells in human colon adenomas and carcinomas, Int.
J. Cancer 73 (1997) 10–15.
[16] M. H .
oythtya, R. Fridman, D. Komarek, et al., Immu-
nohistochemical localization of matrix metalloproteinase
2 and its specific inhibitor TIMP-2 in neoplastic tissues
with monoclonal antibodies, Int. J. Cancer 56 (1994)
500–505.
[17] E.E. Ioachim, A.C. Goussia, M. Machera, et al., Im-
munohistochemical evaluation of cathepsin D expression
in colorectal tumors: a correlation with extracellular
matrix components, p53, pRb, bcl-2, c-erbB-2, EGFR
and proliferation indices, Anticancer Res. 19 (1999)
2147–2156.
[18] B.F. Iskaros, K.E. Tanaka, X. Hu, et al., Morphologic
pattern of tenascin as a diagnostic biomarker in colon
cancer, J. Surg. Oncol. 64 (1997) 98–101.
[19] T. Jahkola, T. Toıvonen, I. Virtanen, et al., Tenascin-C
expression in invasion border of early breast cancer:
a predictor of local and distant recurrence, Br. J. Cancer.
78 (11) (1998) 1507–1513.
[20] M.J. Jessup, M. Loda, Prognostic markers in rectal
carcinoma, Sem. Surg. Oncol. 15 (1998) 131–140.
[21] P.L. Jones, F.S. Jones, Tenascin-C in development and
disease: gene regulation and cell function, Matrix Biol. 19
(2000) 581–596.
[22] A. Juhasz, H. Bardos, G. Repassy, R. Adany, Character-
istic distribution patterns of tenascin in laryngeal
and hypopharyngeal cancers, Laryngoscope 110 (2000)
84–92.
[23] N. Kawano, H. Osawa, T. Ito, et al., Expression of
gelatinase A, tissue inhibitor of metalloproteinase-2,
matrilysin, and trypsin(ogen) in lung neoplasms: an
immunohistochemical study, Hum. Pathol. 28 (5) (1997)
613–622.
B. Sis et al. / Pathology – Research and Practice 200 (2004) 379–387386
ARTICLE IN PRESS
[24] U. Kressner, G. Lindmark, B. Tomasini-Johansson, et al.,
Stromal tenascin distribution as a prognostic marker in
colorectal cancer, Br. J. Cancer 76 (4) (1997) 526–530.
[25] J. Kusukawa, Y. Sasaguri, I. Shima, et al., Expression of
matrix metalloproteinase-2 related to lymph node metas-
tasis of oral squamous cell carcinoma, Am. J. Clin.
Pathol. 99 (1993) 18–23.
[26] A. Mayer, E. Fritz, R. Fortelny, K. Kofler, H. Ludwig,
Immunohistochemical evaluation of cathepsin D
expression in colorectal cancer, Cancer Invest. 15 (2)
(1997) 106–110.
[27] T. Meyer, I.R. Hart, Mechanisms of tumour metastasis,
Eur. J. Cancer 34 (2) (1998) 214–221.
[28] D.L. Ornstein, J. MacNab, K.H. Cohn, Evidence for
tumor-host cooperation in regulating MMP-2 expression
in human colon cancer, Clin. Exp. Metastasis 17 (1999)
205–212.
[29] S.L. Parsons, S.A. Watson, H.M. Collins, et al., Gelati-
nase (MMP-2 and MMP-9) expression in gastrointestinal
malignancy, Br. J. Cancer 78 (11) (1998) 1495–1502.
[30] H. Pilch, U. Schaffer, K. Schlenger, et al., Expression of
tenascin in human cervical cancer-association of tenascin
expression with clinicopathological parameters, Gynecol.
Oncol. 73 (1999) 415–421.
[31] S. Riedl, M. Kadmon, A. Tandara, et al., Tenascin-C
tissue concentration in inflammatory and neoplastic
diseases of the colon mucosa, Anticancer Res. 17 (1997)
3165–3166.
[32] E. Roeb, C.G. Dietrich, R. Winograd, et al., Activity and
cellular origin of gelatinases in patients with colon and
rectal carcinoma differential activity of matrix metallo-
proteinase-9, Cancer 92 (2001) 2680–2691.
[33] S. Sarioglu, E. Ozer, F. Kırımca, et al., MMP-2
expression in laryngeal preneoplastic and neoplastic
lesions, Pathol. Res. Pract. 197 (2001) 483–486.
[34] A. Scorilas, T. Trangas, J. Yotis, C. Pateras, M. Talieri,
Determination of c-myc amplification and overexpression
in breast cancer patients: evaluation of its prognostic
value against c-erbB-2, cathepsin D and clinicopatholo-
gical characteristics using univariate and multivariate
analysis, Br. J. Cancer 81 (8) (1999) 1385–1391.
[35] I. Shima, Y. Sasaguri, J. Kusukawa, et al., Production of
matrix metalloproteinase-2 and metalloproteinase-3 re-
lated to malignant behavior of esophageal carcinoma,
Cancer 70 (1992) 2747–2753.
[36] A. Talvensaari-Mattila, M. Apaja-Sarkkinen, M. H-
.
oyhtya, et al., Matrix metalloproteinase 2 immunoreac-
tive protein appears early in cervical epithelial
dedifferentiation, Gynecol. Oncol. 72 (1999) 306–311.
[37] B. Tetu, J. Brisson, H. Lapointe, Bernard, Prognostic
significance of stromelysin 3, gelatinase A, and urokinase
expression in breast cancer, Hum. Pathol. 29 (9) (1998)
979–985.
[38] G.E. Theodoropoulos, D. Panoussopoulos, A.C. Lazaris,
B.C. Golematis, Evaluation of cathepsin D immunostain-
ing in colorectal adenocarcinoma, J. Surg. Oncol. 65
(1997) 242–248.
[39] J. Upadhyay, B. Shekarriz, J.A. Nemeth, et al., Mem-
brane type 1-matrix metalloproteinase (MT1-MMP) and
MMP-2 immunolocalization in human prostate: change
in cellular localization associated with high- grade
prostatic intraepithelial neoplasia, Clin. Cancer Res. 5
(1999) 4105–4110.
[40] A.M. Valentini, M. Pirrelli, R. Armentano, M.L. Caruso,
The immunohistochemical expression of cathepsin D
in colorectal cancer, Anticancer Res. 16 (1996)
77–80.
[41] B.R. Westley, F.E.B. May, Prognostic value of
cathepsin D in breast cancer, Br. J. Cancer 79 (2) (1999)
189–190.
[42] T. Yoshida, E. Yoshimura, H. Numata, et al., Involve-
ment of tenascin-C in proliferation and migration of
laryngeal carcinoma cells, Virchows Arch. 435 (1999)
496–500.
B. Sis et al. / Pathology – Research and Practice 200 (2004) 379–387 387
