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Expression of Matrix Metalloproteinase-9 mRNA and Vascular Endothelial Growth Factor Protein in Gastric Carcinoma and Its Relationship to Its Pathological Features and Prognosis.

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THE ANATOMICAL RECORD 293:2012–2019 (2010)
Expression of Matrix Metalloproteinase-9
mRNA and Vascular Endothelial Growth
Factor Protein in Gastric Carcinoma and
its Relationship to its Pathological
Features and Prognosis
QIONG YANG,1 ZAI-YUAN YE,1* JING-XIA ZHANG,1 HOU-QUAN TAO,1
SHU-GUANG LI,1 AND ZHONG-SHENG ZHAO2*
1
Key Laboratory of Gastroenterology, Zhejiang Province, Hangzhou, China
2
Department of Pathology, Zhejiang Provincial People’s Hospital, Hangzhou, China
ABSTRACT
To investigate matrix metalloproteinase-9 (MMP-9) mRNA and vascular endothelial growth factor (VEGF) protein expression in gastric carcinoma and its correlation with microvascular density, growth-pattern,
invasion, metastasis, and prognosis. In situ hybridization of MMP-9 mRNA
and immunohistochemistry of VEGF and CD34 proteins were performed
on surgical specimens of gastric cancers from 118 patients compared with
20 nonmalignant gastric mucosae. Their relationships to pathological parameters and survival times were determined by statistical analysis. The
positive rate of MMP-9 in noncancerous gastric mucosae was significantly
lower than that of gastric cancer tissue (60.17%, P < 0.01). In patients
with cancers of the infiltrating type, at stage T3-T4, with vessel invasion,
lymphatic metastasis, hepatic, or peritoneal metastasis, the positive
expression rates of MMP-9 mRNA, VEGF protein, and CD34 were significantly higher than those for patients with tumors of the expanding type (P
< 0.01), at stage T1–T2 (P < 0.01), with nonvessel invasion (P < 0.05),
without lymphatic metastasis (P < 0.05), and without hepatic (P < 0.001)
or peritoneal metastasis (P < 0.001), respectively. Expression of MMP-9
mRNA was positively related to that of VEGF protein (P < 0.001) and microvascular density (P < 0.001). Patients with higher MMP-9 mRNA and
VEGF expression demonstrated vivid tumor angiogenesis and poor 5-year
survival rate. MMP-9 and VEGF expression is associated with enhanced
tumor angiogenesis and may play crucial roles in the invasion and metastasis of gastric carcinoma. Therefore, MMP-9 and VEGF may represent
prognostic biomarkers and promising targets for therapeutic intervention.
C 2010 Wiley-Liss, Inc.
Anat Rec, 293:2012–2019, 2010. V
Key words: stomach neoplasms; matrix metalloproteinase-9;
vascular endothelial growth factor; metastasis;
prognosis
Grant sponsor: Zhejiang Province Natural Science
Foundation; Grant number: M303843.
*Correspondence to: Zai-Yuan Ye, Key Laboratory of Gastroenterology, Zhejiang Province, Hangzhou 310014, Zhejiang,
China. E-mail: zaiyuanye@163.com or Z.-S. Zhao, Department
of Pathology, Zhejiang Provincial People’s Hospital, Hangzhou
310014, Zhejiang, China. E-mail: zhongshengzhao@163.com
C 2010 WILEY-LISS, INC.
V
Received 1 April 2009; Accepted 8 September 2009
DOI 10.1002/ar.21071
Published online 18 November 2010 in Wiley Online Library
(wileyonlinelibrary.com).
EXPRESSION OF (MMP-9) mRNA AND VEGF PROTEIN
INTRODUCTION
The incidence and mortality rates of gastric cancer
have fallen dramatically in the past 70 years. Despite its
decreasing prevalence, gastric carcinoma remains the
second leading cause of cancer death worldwide (Crew
and Neugut, 2006). Most patients are diagnosed late at
the mid-advanced stage, resulting in an overall 5-year
survival rate >20%. Tumor cells degrade extracellular
matrix (ECM) components to invade surrounding tissues. The process is precisely controlled by ECM-degrading enzymes including matrix metalloproteinases
(MMPs). MMP-9 is a zinc-containing enzyme with potent
proteolytic activity against a wide range of ECM components such as laminin-5 and type IV collagen, the major
constituents of basement membranes. Recent evidence
has uncovered multiple functions for MMP-9, in addition
to simply degrading ECM, which may partially contribute to tumor angiogenesis and metastasis (Hiratsuka
et al., 2002).
Aberrantly expressed MMP-9 has been reported in a
variety of tumors. However, its clinical role in gastric
cancer is still fragmentary. Thus, to explore its malignant influence and prognostic significance, we examined
the expression of MMP-9 mRNA and vascular endothelial growth factor (VEGF) and microvascular density
(MVD) in 118 tissue samples from gastric cancer
patients. The correlation between MMP-9 and tumor
angiogenesis and other clinical pathological parameters
of gastric carcinomas including growth-pattern, invasion,
and metastasis were also analyzed in this study.
MATERIALS AND METHODS
Patients and Specimens
A total of 118 patients (79 male, 39 female; aged 38–
80 years, median age 57.8 years) who underwent gastrectomy for gastric carcinoma at Zhejiang Provincial
People’s Hospital from October 1990 to November 1998
were included in this study. Five-year follow-up data
was obtained, and the follow-up ended in November
2003. According to the World Health Organization classification standard (2002), there were 39 tubular adenocarcinomas, 19 papillary adenocarcinomas, 37 poorly
differentiated adenocarcinomas, 12 mucinous adenocarcinomas, and 11 signet ring cell carcinomas. These cancerous tissues of these patients were also classified into
well- and moderately-differentiated (G1 þ G2, 70
patients) and poorly-differentiated (G3 þ G4, 48
patients) types, based on the predominant differentiation
mode, and further classified into expanding (51 patients)
and infiltrating (67 patients) types. According to the
AJCC TNM staging system (Greene et al., 2002), there
were 21 cases of stage T1, 26 cases of stage T2, 45 cases
of stage T3, and 26 cases of stage T4. There were 89
patients with and 29 without vessel invasion, 84
patients with lymphatic metastasis and 34 without, 55
patients with distant metastasis (35 with peritoneal dissemination, 20 with hepatic metastasis) and 63 without.
A control study was carried out on 20 samples obtained
from adjacent noninvolved normal gastric mucosa 5 cm
away from the primary tumor without hyperplasia or
atypical hyperplasia.
2013
Reagents
MMP-9 (MK1540) probe: Digoxin-labeled nucleotides
for MMP-9 probes were obtained from Boster Biological
Technology Limited Company, Wuhan, China. The target
gene mRNA sequences were as follows: (1) 50 -TCCCT
GCCCG AGACC GGTGA GCTGG ATAGC-30 ; (2) 50 CAACT CGGCG GGAGA CTGT GCGTC TTCCC-30 ; and
(3) 50 -CCAGG TGGAC CAAGT GGGCT ACGTG ACCTA30 . Immunohistochemical reagents including primary
monoclonal antibodies against VEGF (1:100) and CD34
(1:120) and EnVision kit were obtained from DAKO Denmark. (Produktionsvej 42 DK-2600 Glostrup). The VEGF
clone, JH121, and the CD34 clone, QBEnd 10, were
used.
In Situ Hybridization
In situ hybridization was performed on formalin-fixed
and paraffin-embedded tissue sections according to the
manufacturer’s instructions. Samples cut at 4-lm intervals were dewaxed in xylene and rehydrated. The endogenous peroxidase was blocked with 3% hydrogen
peroxide. After washing three times with distilled water,
the sections were digested at 37 C with Pepsin Reagent
for 30 min, fixed with 1% paraformaldehyde plus 1/1,000
DEPC for 10 min, and then incubated in 50-lL prehybridization solution at 40 C for 4 hr. Each slide was
then incubated with 20-lL hybridization solution
(hybridization probe concentration ¼ 2 ng/lL) overnight
in a humidified chamber at 40 C, washed twice with 2
SSC, once with 0.5 SSC, and finally three times with
0.2 SSC. After that blocking solution was applied and
the superfluous liquid was absorbed. Slides were then
treated with mouse biotinylated antidigoxigenin for
60 min, washed three times with PBS, and then incubated in SABC for 30 min. The hybridization signal was
detected by using the avidin-biotin-peroxidase technique
with DAB as the chromogen. The slides were counterstained with Harris hematoxylin, washed, dehydrated in
ethanol, cleared in xylene, and enveloped with neutral
gum. Negative controls included the following: (1)
hybridization solution without probe, and (2) specimens
pretreated with RNase.
Immunohistochemistry
The EnVision two-step method was performed according to the manufacturer’s instructions. Tissue sections
were cut at 4-lm intervals, dewaxed with xylene,
hydrated by a series of ethanol solutions (100%, 95%,
80%, and 70%). High temperature and high pressure
(CD34: 0.01 mol/L sodium citrate buffering solution, pH
6.0; VEGF: 0.01 mol/L EDTA buffering solution, pH 9.0)
were applied to facilitate antigen retrieval. The sections
were washed once with distilled water, three times with
PBS for 5 min, and with inactivated endogenous peroxidase using 3% hydrogen peroxide for 10 min at room
temperature. After washing three times with PBS for
5 min, sections were incubated overnight with the primary antibody at 4 C. They were rinsed three times
with PBS for 5 min and incubated with goat anti-mouse
IgG antibody/HRP polymer for 40 min at 37 C before
washing three times with PBS for 5 min. Visualization
was achieved with DAB incubation for 3 min. After
counterstaining with Harris hematoxylin, the slides
2014
YANG ET AL.
were dehydrated in 95% and 100% ethanol, cleared in
xylene, and enveloped with neutral gum. The negative
control included replacement of the primary antibody
with PBS, and the positive slides provided by the reagent kit were used as the positive control.
Results Evaluation
Cytoplasm stained brown was indicative of the positive expression of MMP-9 mRNA. The staining results
ranging from to þþþ were estimated as follows: ()
(positive cell numbers <10% or no staining); (þ) (positive
cell numbers 11%–50%); (þþ) (positive cell numbers
51%–75%); or (þþþ) (positive cell numbers >75%). Signals for VEGF expression was detected as brown in the
cytoplasm and/or cell membrane. Based on the positive
cell counting, the results were scored as follows: () (no
staining); (þ) (positive cell numbers <25%); (þþ) (positive cell numbers 26%–50%); or (þþþ) (positive cell
numbers >50%). Calculation of MVD was performed as
well. Vessels with a clearly defined lumen or welldefined linear vessel shapes, but not single endothelial
cells, were considered for microvascular assessment.
Each slide was first examined at 100 magnification to
capture five ‘‘hot spots,’’ defined as areas with highest
vascularization. CD34þ vessels were then quantified in
those chosen fields (200). The average count was taken
as MVD and statistically presented as the mean SD.
The patients were divided into two groups as high MVD
(54.9/mm2) and low MVD (<54.9/mm2).
Statistical Analysis
Data were analyzed statistically using SPSS 13.0 software. Significant differences were evaluated with Student’s t-tests. The v2 test was performed on the
numerical data. Survival analysis was carried out using
the Kaplan-Meier product-limit method, and survival
curves were plotted. The differences were evaluated by
the log rank test. A P-value <0.05 was considered statistically significant.
RESULTS
Correlation Between MMP-9 mRNA Expression
and Clinical Pathological Parameters
Negative results were found in normal gastric mucosae (Fig. 1A), whereas 71 of the 118 (60.7%) gastric cancer samples, including those in early stages, showed
positive MMP-9 expression. Signals of positive staining
were detected in the cytoplasm of the carcinoma cells,
which was usually located near the edge of the carcinoma filtration areas. Most of the greater omentum and
peritoneum were also positive for MMP-9 expression
(Fig. 1B). In patients with the infiltrating type of cancer,
at stage T3–T4, with vessel invasion, lymphatic metastasis, hepatic, and peritoneal metastasis, the positive
expression rates of MMP-9 mRNA was significantly
higher than for those patients with the expanding type
(v2 ¼ 8.513, P < 0.01), at stage T1–T2 (v2 ¼ 12.706, P <
0.001), without vessel invasion (v2 ¼ 5.664, P < 0.05),
without lymphatic metastasis (v2 ¼ 8.446, P < 0.05), or
without hepatic and peritoneal metastasis (v2 ¼ 17.378,
P < 0.001; v2 ¼ 21.938, P < 0.001). No statistical correlation was revealed between MMP-9 mRNA positive
Fig. 1. In situ hybridization expression of MMP-9 mRNA in samples
from gastric adenocarcinoma patients: (A) MMP-9 mRNA is negative
in adjacent normal gastric mucosa (magnification 200); (B) Cytoplasm of gastric cancer mucosa demonstrates high positive rates of
MMP-9 mRNA (magnification 200).
expression and the carcinoma differentiation (v2 ¼
1.188, P ¼ 0.276; Table 1).
Correlation Between VEGF Expression and
Clinical Pathological Features
VEGF was rarely expressed in normal gastric mucosae
(Fig. 2A). Whereas of 118 gastric carcinoma patients, 64
positively expressed VEGF (54.2%). The staining signal
was mainly located in the cytoplasm of tumor cells and
usually observed at the invasive edge of tumor cell nests
(Fig. 2B). In patients with the infiltrating type of
tumors, at stage T3–T4, with vessel invasion, lymphatic
metastasis, hepatic, and peritoneal metastasis, the
VEGF positivity was much more prevalent compared
with those for patients with the expanding type (v2 ¼
10.18, P < 0.01), at stage T1–T2 (v2 ¼ 34.19, P < 0.001),
with nonvessel invasion (v2 ¼ 37.29, P < 0.001), without
lymphatic metastasis (v2 ¼ 34.71, P < 0.001), and without hepatic or peritoneal metastasis (v2 ¼ 24.53, P <
0.001; v2 ¼ 49.75, P < 0.001). However, no correlation
was found between the different histological types (v2 ¼
3.49, P ¼ 0.062; Table 1).
2015
EXPRESSION OF (MMP-9) mRNA AND VEGF PROTEIN
TABLE 1. Correlation of MMP-9 mRNA, VEGF protein, and MVD with clinical pathological
characteristics
VEGF
MMP-9
Growth pattern
Expansive
Infiltrative
Histological grade (G)
G1 þ G2
G3 þ G4
Invasive depth
T1–T2
T3–T4
Vessel invasion
No
Yes
Lymph node metastasis
No
Yes
Distant metastasis
No
Liver
Peritoneal dissemination
N
þ
þþþ
118
51
67
47
28
19
71
23
48
70
48
32
15
38
33
47
71
28
19
19
52
29
89
17
30
12
59
34
84
21
26
13
58
63
20
35
40
2
5
23
18
30
P
X2
8.51
<0.01
1.19
>0.05
12.71
5.66
8.45
31.58
17.38a
21.94b
<0.001
<0.05
<0.05
<0.001
<0.001a
<0.001b
þ
þþþ
54
32
22
64
19
45
37
17
33
31
37
17
10
54
28
26
1
63
30
24
4
60
50
3
1
13
17
34
X2
P
10.18
<0.01
3.49
>0.05
34.19
<0.001
37.29
<0.001
34.71
<0.001
61.49
<0.001
24.53a
49.75b
<0.001a
<0.001b
MVD (/mm2)
49.45 21.72
64.06 18.76
55.72 21.56
60.70 20.73
44.42 19.96
66.56 17.23
35.68 14.04
64.93 18.07
37.60 15.73
66.24 17.24
42.81 17.23
73.40 8.07
75.79 9.48
t
P
3.92
<0.001
1.25
>0.05
6.41
<0.01
7.96
<0.01
8.45
<0.001
7.65a
10.46b
<0.01a
<0.01b
a
Compared between patients with liver metastasis and those without distant metastasis.
Compared between patients with peritoneal dissemination and those without distant metastasis.
b
Correlation Between MVD and Clinical
Pathological Features
Correlation Between MMP-9 mRNA, VEGF
Protein, MVD, and Prognosis
CD34 staining was positive in vascular endothelial
cells. Microvessels in cancerous tissue and surrounding
areas were intensely stained from brown to dark brown.
In patients with the infiltrating type of cancer, at stage
T3–T4, with vessel invasion, lymphatic metastasis, hepatic, and peritoneal metastasis (Fig. 3A), the mean
MVDs were significantly higher than those for patient
with the expanding type (t ¼ 3.92, P < 0.001), at stage
T1–T2 (t ¼ 6.41, P < 0.01), with nonvessel invasion (t ¼
7.96, P < 0.01), without lymphatic metastasis (t ¼ 8.45,
P < 0.001), and without hepatic or peritoneal metastasis
(t ¼ 7.65, P < 0.01; t ¼ 10.46, P < 0.01), respectively
(Fig. 3B). No correlation was found within the different
histological types (P > 0.05; Table 1).
The survival time in patients positive for the expression of MMP-9 mRNA and VEGF protein with an MVD
54.9/mm2 was significantly shorter than for patients
who were negative for the expression of MMP-9 mRNA
(P < 0.001) and VEGF protein (P < 0.01) with an MVD
< 54.9/mm2 (P < 0.001). The 5-year survival rate in
patients positive for the expression of MMP-9 mRNA
and VEGF with an MVD 54.9/mm2 was significantly
lower than patients negative for expression of MMP-9
mRNA (P < 0.001) and VEGF protein (P < 0.01) with an
MVD < 54.9/mm2 (P < 0.001; Figs. 4–6; Table 2).
Correlation Between MMP-9 mRNA, VEGF
Protein Expression, and MVD
The mean MVD (64.28 19.54/mm2) in gastric carcinoma specimens with a positive expression of MMP-9
mRNA was much higher than in the negative expression
group (47.88 20.13/mm2; t ¼ 4.410, P < 0.001). Similarly, the mean MVD in gastric carcinoma specimens
positive for the expression of VEGF (71.98 13.26/mm2)
was higher than in the negative expression group
(39.678 14.81/mm2; t ¼12.475, P < 0.001). In addition,
the percentage of VEGF expression in carcinoma tissues
in conjunction with a positive expression of MMP-9
mRNA was 73.2% (52/71), which was significantly higher
than the MMP-9 mRNA negative expression group
(25.5%, 12/47; P < 0.001). Therefore, there was a positive relationship between the expression of MMP-9
mRNA, VEGF protein, and MVD.
DISCUSSION
MMP-9 has a wide range of proteolytic activities
including the capacity to degrade ECM components.
Similar to other members of the family, function of
MMP-9 is mainly based on its multidomain structure,
which includes a highly conserved catalytic domain,
HEXXHXXGXXH. This domain depends on Zn2þ for
activation, a C-terminal hemopexin-like domain for collagenase digestion, and three additional repeats of the fibronectin type II motif located in the metalloproteinase
domain for the determination of substrate specificity
(Nagase et al., 2006).
Imbalanced secretion of MMP-9 and its endogenous
inhibitors into the surrounding microenvironment during cancer progression may alter the noncellular compartment, causing tissue remodeling and facilitating
tumor cell migration. Aside from its enzymatic activities,
MMP-9 also affects cytoskeletal organization through
interactions with different families of adhesion receptors
and regulates vessel stabilization by controlling pericyte
recruitment (Bendeck et al., 2000; Chantrain et al.,
2016
YANG ET AL.
Fig. 2. Immunohistochemical expression of VEGF in samples from
gastric adenocarcinoma patients: (A) VEGF protein is slightly
expressed in normal gastric mucosa (magnification 400); (B) Cytoplasm of gastric cancer mucosa demonstrates a high expression for
VEGF protein (magnification 400).
2004). Associations between MMP-9 expression levels
with invasive phenotypes have been increasingly recognized in tumors of different anatomical origins. For
example, oncogenes H-ras and v-myc transformed into
rat embryo cell lines were proved to be highly metastatic
and over-expressed MMP-9 (Himelstein et al., 1997).
Cells derived from human lung cancer treated with GCSF, GM-CSF, M-CSF demonstrate an enhanced production of specific uPA and MMP-9, which was responsible
for the subsequent increases in metastatic potential (Pei
et al., 1999). Blocking the ERK1/2 or p38-MAPK pathway
alone contributes to a decrease in MMP-9 expression
and subsequently suppresses the malignancy of renal
cancer or squamous cell cancer (Simonc et al., 1998;
Hong et al., 2005). Recently, Roelle et al. (2003) have
confirmed for the first time an expanded role for MMP-9
in the endocrine promotion of pituitary tumors (Roelle
et al., 2003). According to their research, the GnRHinduced Src, Ras, and ERK activation and the enhanced
crosstalk between GnRHR and EGFR in gonadotropic
cells were gelatinase-dependent on the c-Jun NH2-terminal kinase, but not the extracellular signal-regulated
kinase1/2 or p38-MAPK. The signaling pathway was sup-
Fig. 3. Immunohistochemical expression of CD34 in samples from
gastric adenocarcinoma patients: (A) Immunohistochemical expression
of CD34 in vascular endothelial cells in gastric carcinoma when MVD
54.90/mm2 (magnification 200); (B) Immunohistochemical expression of CD34 in vascular endothelial cells in gastric carcinoma when
MVD <54.90/mm2 (magnification 200).
posed to be critical for GnRH-mediated upregulation of
MMP-9, cell invasion, and motility. On the other hand,
the metastasis-association hypothesis was supported by
other studies of gene-expression signatures, which suggested MMP-9 was among the 70 genes comprising a
gene signature that are capable of predicting distant metastasis in lymph node-negative breast cancer patients
(van’t et al., 2002). Likewise, gastric carcinomas coincide
with a notably upregulated MMP-9 expression. In fact,
poorer survival rates were found in patients with elevated plasma MMP-9 levels compared with patients
with normal MMP-9 plasma levels. However, MMP-9 in
serum did not correlate well with the invasiveness or
survival time of gastric cancer (Wu et al., 2007).
As uncontrolled ECM remodeling of basement membranes is a common feature of metastasis, we compared
MMP-9 mRNA expression in human gastric cancer tissues with nontumor mucosae using in situ hybridization.
A significantly enhanced production of MMP-9 was
observed in cancer tissues, especially those tumors characteristic of the infiltrating type, at stage T3–T4, with
vessel or lymphatic invasion, and with hepatic or
EXPRESSION OF (MMP-9) mRNA AND VEGF PROTEIN
Fig. 4. Survival curves with positive and negative MMP-9 mRNA
expression in gastric adenocarcinoma (P < 0.001).
Fig. 5. Survival curves with positive and negative VEGF expression
in gastric adenocarcinoma (P < 0.001).
peritoneal metastasis. This aberrant proteinase upregulation may represent an important biological mechanism
responsible for the increasing malignant behavior of gastric cancer.
Tumors require angiogenesis to continue to grow. One
critical event implicated in the migration and proliferation of vascular endothelial cells is the proteolytic degradation of basement membranes and ECM components by
matrix metalloproteinases (MMPs). In this study, MMP9 mRNA in the gastric cancer tissues was upregulated
and coincided with VEGF expression and microvascular
density. These findings support an emerging notion that
MMP-9 is a positive regulator of tumoral angiogenesis
(Cox et al., 2000). Hawinkels et al. (2008) showed that
2017
Fig. 6. Survival curves with MVD < 54.90/mm2 and 54.90/mm2 in
gastric adenocarcinoma (P < 0.001).
neutrophil-derived MMP-9 was able to release the biologically active VEGF165 from the ECM of colon cancer
once liberated from heparan sulfates (Hawinkels et al.,
2008). Heissig et al. (2002) found MMP-9/ mice given
5-FU treatment showed a delayed hematopoietic recovery compared with the MMP-9þ/þ mice. MMP-9 secreted
within the bone marrow altered the stem cell-stromal
cell interaction in the microenvironment. These effects
of MMP-9 have been shown to be enhanced by SDF-1,
VEGF, and G-CSF (Lévesque et al., 2003; Xu et al.,
2005). The cleavage of vascular cell adhesion molecule-1,
expressed by BM stromal cells, and the release of the
soluble c-kit ligand from a membrane bound state, has
been largely implicated in promoting stem cell differentiation, accelerating hematopoietic reconstitution, as
well as enhancing their mobilization into peripheral circulation. Consequently, these properties suggest a key
role for the proteolytic cleavage of some cytokines in vasculogenesis (Jeon et al., 2007).
In conclusion, our study identified that gastric cancer
mucosae produced a significantly higher level of MMP-9
mRNA compared with healthy ones. The positive rates
of MMP-9 expression were remarkably elevated in tumor
features associated with the infiltrating type, at stage
T3–T4, with vessel invasion, lymphatic metastasis, and
hepatic or peritoneal metastasis when compared with
the expanding type of gastric cancer, at stage T1–T2,
with nonvessel invasion, without lymphatic metastasis,
without hepatic and peritoneal metastasis. Tumors with
different differentiations did not generate such a distinction. On the other hand, evidence from immunohistochemistry revealed a positive correlation between the
expression profiles of MMP-9, VEGF, and CD34, representing a vivid tumor vascular growth pattern. To
explore the survival influence of those molecules,
Kaplan–Meier survival analysis was then performed.
The curves indicated that gastric tumors expressing
high levels of MMP-9 mRNA, and VEGF and CD34
protein were more inclined to become invasive.
2018
YANG ET AL.
TABLE 2. Correlation of MMP-9 mRNA, VEGF protein, and MDV with survival period
N
MMP-9 mRNA
VEGF
MVD
þþþþ
þþþþ
<54.90
54.90
47
71
54
64
47
71
Mean survival
period (months)
88.38
40.24
114.53
32.13
122.61
30.58
6.37
5.95
6.66
4.25
6.63
3.75
Consequently, patients with these types of tumors had a
poorer 5-year survival rate.
In light of the earlier observations, we believe MMP-9
and VEGF may serve as useful molecular biomarkers to
predict the aggressiveness of gastric carcinoma and to
determine the prognosis of tumors which express it.
Therapeutic interventions aimed at this two promising
targets are worth exploring in future longitudinal studies. However, most clinical trials of anti-MMPs compounds to date have failed to obtain satisfactory effects.
Even so, we can still draw some inspirations from the
updated attempts. For example, In the study of Kunigal,
antisense Ad-MMP-9 were successfully applied to
decrease the levels of transcription factors nuclear factor—kappa B and activator protein, both of which participated in the triggering of the Fas-Fas ligand
apoptotic cascade, and finally overcame the radiotherapy
resistance of breast cancer (Kunigal et al., 2008). Thus,
to solve this problem of poor response, multiple transcription factor consensus binding motifs in the regulatory regions, including those for SP-1, Ets, AP-1, and
REB, might be the logical candidates for targeting
(Himelstein et al., 1997; Ma et al., 2001).
Taken together, MMPs is such a large family that the
functions of which are so complex that far beyond our
current understanding. Considering that expression of
MMPs is mainly controlled at the level of transcription,
and a number of transcriptional sites have already been
characterized now (Ahn et al., 2008; Kim et al., 2008).
Novel strategies to target this type of proteinase more
selectively or to design drugs against critical transduction signals or key gene expression during the progression of gastric cancer may provide some advantages for
the survival of patients with gastric carcinoma.
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expressions, features, carcinoma, matrix, pathologic, growth, vascular, gastric, relationships, prognosis, mrna, factors, endothelial, protein, metalloproteinase
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