Original Articles
Matrix metalloproteinase-1 expression in betel quid-associated
oral cancers
CHANG-TA CHIU1,2,3
SHENG-YANG LEE3
WEI-FAN CHANG5
CHING-YU YEN5
DUAN-JANG WANG3
YOUNG-CHAU LIU4
CHIN-HAI LEE5
SHYUN-YEU LIU3,5
1
Department of Oral and Maxillofacial Surgery, Chang Gung Memorial Hospital-Kaohsiung Memorial Center, Kaohsiung,
Taiwan, ROC.
2
Chang Gung University, College of Medicine, Taoyuan, Taiwan, ROC.
3
School of Dentistry, Taipei Medical University, Taipei, Taiwan, ROC.
4
College of Liberal Education, Shu-Te University, Kaohsiung, Taiwan, ROC.
5
Department of Oral and Maxillofacial Surgery, Chi Mei Medical Center, Liouying, Tainan, Taiwan, ROC.
Approximately 80%~90% of Taiwanese patients with oral cancer have chewed betel nuts for a long
time. Some ingredients of the nuts may mechanically and chemically disrupt the oral mucosa. Matrix
metalloprotinase-1 (MMP-1) is considered to be important in degrading the extracellular matrix. Recent
studies have shown that the overexpression of MMP-1 is associated with the occurrence, proliferation,
and prognosis of oral cancer as well as lymph node metastasis. Most published studies related to MMP-1
have involved patients with oral cancer from outside of Taiwan, and few have concentrated on patients
from Taiwan, who are often habitual betel nuts chewers. Therefore, this research, is aimed at patients in
Taiwan, analyzed the MMP-1 expression of oral cancer of the patients. The Department of Oral and
Maxillofacial Surgery of Chi-Mei Medical Center specifically studied 30 patients with oral cancer and a
history of betel nut chewing and their relation to MMP-1 expression. In the immunohistochemical
staining analysis, MMP-1 expression was higher in the epithelium of tumor tissues in 19 of 30 patients
(63.3%). After further statistical analysis (Fisher’s exact test), MMP-1 expression has shown to be
stronger in late stages (stages III and IV) than in early stages (stages I and II). In addition, MMP-1
expression also increased with increasing lymph node metastasis staging (from N0 through N1 to N2) and
differentiation grade (from G1 through G2 to G3). These results prove the significant role of MMP-1 in
betel nut chewing-associated oral cancer, tumor growth, tumor differentiation, and lymph node
metastasis. （J Dent Sci, 3(2)：75-82 , 2008）
Key words: oral cancer, betel nut, metalloproteinase-1, immunohisto-chemical staining.
Oral cancer is common in the world, especially in
Taiwan. Its mortality rate ranks sixth among cancers
in Taiwan; it is rated second in the annual increase
in the mortality rate. The rate of squamous cell
carcinoma (SCC) among oral cancers is the highest
Received: February 15, 2008
Accepted: May 20, 2008
Reprint requests to: Dr. Shyun-Yeu Liu, Department of Oral and Maxillofacial Surgery, Chi-Mei Hospital, Liouying, No.
201, Taikang Village, Liouying Township, Tainan
County, Taiwan 73657, ROC.
J Dent Sci 2008‧Vol 3‧No 2
(95%); and 80%~90% of SCC patients in Taiwan have
the habit of chewing betel nuts (Areca catechu); and
60%~70% of them continue chewing betel nuts after
the cancer has been diagnosed. In the College of Oral
Medicine, Kaohsiung Medical University, betel quid
was examined and had high mutagenic potency; its
main extracts are sugars, polyphenols, tannins, and
alkaloids1. These ingredients can hurt the oral mucosa
mechanically and chemically. Patients may have the
following clinical symptoms: (a) ulcerated, atrophic,
thickened epithelium; (b) epithelial color changes; (c)
oral submucosal fibrosis and a burning sensation;
75
C.T. Chiu, S.Y. Lee, D.J. Wang, et al.
and (d) desquamative and pseudo-membranous or
wrinkle-like changes2. The histological expression
in patients’ oral tissues can include: (a) brownish
amorphous substance-covered irregular epithelium; (b)
a ballooning appearance due to granular materials in
cells; (c) epithelial hyperplasia with significant rete
pegs and massive inflammatory cell infiltration under
the epithelium; (d) an increase in epithelial cell
division ability; (e) basal nuclei hyperkeratosis and
pyknotic changes; (f) probable cell dysplasia changes;
and (g) decreases in types III and IV collagen proteins
in fibrous connective tissue, with an increase in type I
collagen protein3. Betel nut cytotoxicity can induce
DNA injury, inhibit DNA repair, and eventually
induce differentiation of keratinocytes of oral mucosal
cells4. Studies have reported that betel nuts mainly
destroy keratinocytes and fibroblasts. Inflammation
associate with the former cells is important during
tumor changes, and betel quid extract (BQE) can
induce the release of PGE2 and 6-keto-PGF1α by
oral keratinocytes, both of which result in local
inflammation of membranous cells5. BQE-stimulated
keratinocytes are depleted of the antioxidant glutathione, thus easily giving rise to epithelial cancer6.
During betel nut chewing, the mucosal membranes
contact betel nut alkaloids with different densities and
multiple types of nitrosamines which can be a basic
factor in oral mucosal lesions and a carcinogen7. Areca
nuts contain arecoline and arecaidine which both can
cause mutations.
Matrix metalloproteinases (MMPs), a group of
zinc-dependent endopeptidases, are involved in
exreacellular matrix (ECM) physiological degradation
processes, like embryonic development, angiogenesis,
and wound healing8,9. Moreover, MMPs influence
other pathological processes of diseases, like those
of rheumatoid arthritis, periodontal diseases, autoimmune diseases, osteoarthritis, tumor invasion, and
metastasis10. During tumor invasion, cancer cells
break through the basement membrane and enter
connective tissue, which is a crucial basic characteristic, and varies with changes in the interactions of cells, matrix, and matrix-degrading
enzymes, among which MMPs are the most
important10,11. Among tumor cells, inflammatory cells
are characterized by their potential for infiltrating
tissues. Inflammatory cells not only release MMPs to
the peripheral region of a tumor but also release
cytokinase to activate MMPs12. In malignant tumors,
stromal fibroblasts are the main source of MMPs13.
76
Therefore it would be useful to investigate the
regulation of MMPs in tumor cells in order to
understand the mechanisms of invasion and metastasis
of tumor cells.
Collagenases are MMPs that mainly degrade
types I, II, and III collagen in connective tissues. At
present, collagenase-type MMPs include MMP-1
(collagenase-1), MMP-8 (collagenase-2), and MMP13 (collagenase-3). MMP-1 is also known as
fibroblast or interstitial collagenase, and many cells,
including fibroblasts, macrophages, keratinocytes, and
epithelial cells, express MMP-1. It is also expressed
by basal keratinocytes at wound margins14. MMP-1
expression is upregulated in colorectal cancer15, lung
cancer16, and esophageal cancer17. Kurahara et al.18
showed that MMP-1 overexpression is associated with
the proliferation of oral neoplasms. Sutinen et al.19,20
demonstrated that MMP-1 expression is related to the
development of oral cancer and its prognosis.
O-Charoenrat et al.21 reported that MMP-1 expression
is increased in patients with oral neoplasia, and
that it is related to lymph node involvement. Immunohistochemical (IHC) studies have revealed that
MMP-1 is related to differentiation of epithelial cells
in oral cancer, in addition to activation of the MMP-1
gene, which can induce invasion in an oral cancer cell
line22.
With this information from the literature and
other related studies, a close relationship appears to
exist between MMP-1 expression and oral cancer.
Particularly with oral cancer, migration of epithelial
cells into the underlying connective tissue is an
important characteristic of tumor invasion. Degradation of the ECM, which is abundant in oral
submucosal connective tissue, is important in the
progression of oral squamous cell carcinoma.
Therefore, MMP-1, which mainly participates in ECM
degradation, may be useful in serving as a cellular
marker of oral cancer and in estimating its prognosis23.
Most published studies related to MMP-1 have
involved patients with oral cancer from outside
Taiwan, and few have concentrated on patients from
Taiwan, most of whom habitually chew betel nuts. In
the oral cavity, constituents of betel nuts mainly
damage keratinocytes and fibroblasts, potentially
inducing further abnormal MMP-1 secretion by those
cells. Therefore, the purpose of our study was to
analyze the relationships of MMP-1 expression with
the progression, cellular differentiation, and lymph
node involvement of oral cancers in patients from
J Dent Sci 2008‧Vol 3‧No 2
MMP-1 expression in oral cancers
Taiwan, where betel nut chewing is prevalent.
MATERIALS AND METHODS
Patients
We included 30 patients with oral cancer that
were surgically treated at the Department of Oral and
Maxillofacial Surgery, Chi-Mei Medical Center.
Patients had no other systemic diseases and denied
smoking and drinking habits, but all had a >10-year
history of betel nut chewing. Their cancers involved
the buccal mucosa (n=12), tongue (n=7), lower
gingiva (n=6), mouth floor (n=3), retromolar trigone
(n=1), and soft palate (n=1). Stages of cancer were I
(n=7), II (n=6), III (n=3), and IV (n=14). According to
the tumor sizes, lymph node states, and distal
metastases (TNM) classification, stages were N0
(n=18), N1 (n=4), and N2 (n=8). Tumor tissue was
well differentiated (grade G1) in 13 patients, moderately differentiated (G2) in 13, and poorly differentiated (G3) in 4 (Table1).
Immunohistochemistry staining
Blocks of tumor tissue were obtained from the
study subjects and embedded and processed in our
Department of Pathology. The above study was
approved by the Institutional Review Board (IRB) of
Chi-Mei Medical Center and was monitored by the
IRB. Samples were cut into 5-µm slices using a
microtome and dried flat on microscope slides before
staining. Each slide was deparaffinized using xylene
and high- to low-concentration ethanol, after which it
was boiled in a 10 mM sodium citrate solution (pH 6.4)
for 30 minutes to expose the tissue antigens. Slides
were reacted in H2O2 for 10 minutes before being
washed with 1× phosphate-buffered saline (PBS). We
then added 1% of an MMP-1 primary antibody (clone
EP1247Y; Epitomics, Burlingame, CA), which was
reacted at room temperature for 2.5 hours or at 4 ºC
overnight. After washing the slide with 1× PBS, we
added a biotinylated secondary antibody and allowed
the reaction to proceed at room temperature for 30
minutes. The slides were washed again with 1× PBS
buffer, before streptavidin peroxidase was added and
reacted at room temperature for 30 minutes.
The substrate, 3-amino-9-ethylcarbazole, was
used for chromogenesis, and hematoxylin was used
for counterstaining. Slides were mounted using
mounting medium, then incubated and dried at 60 ºC
for 30 minutes. Finally, 1 pathologist examined
MMP-1 expression in the tumor tissues under 400×
light microscopy, and photomicrographs were taken
for later reference. The cutoff point for all-or-none
expression was defined as expression in ≥10% of
epithelial cells.
Statistical analysis
Fisher’s exact test was used to determine whether
MMP-1 expression had increased in stage I and II
cancer vs. stage III and IV cancer. Fisher’s exact test
was also used to determine whether the expression of
MMP-1 increased with involvement of lymph nodes
Table 1. Sites, stages, differentiation states, and sizes of the 30 oral squamous cell carcinoma patients
Differentiation
Site
Stage
State
Size
BM
12/30
Ⅰ
7/30
Well
13/30
T1
8/30
G
6/30
Ⅱ
6/30
Moderate
13/30
T2
9/30
T
7/30
Ⅲ
3/30
Poor
4/30
T3
2/30
MF
3/30
Ⅳ
14/30
T4
11/30
SP
1/30
R
1/30
BM, buccal mucosa; G, gingival; T, tongue; MF, Mouth floor; SP, soft palate; R, retromolar trigon.
J Dent Sci 2008‧Vol 3‧No 2
77
C.T. Chiu, S.Y. Lee, D.J. Wang, et al.
metastasis from stages N0 to N1 and N2. After
conducting a Kruskal-Wallis H test then post hoc
comparisons, we applied Dunn’s test to determine
whether MMP-1 expression was stronger in patients
with G2 and G3 disease than in those with grade G1
disease.
RESULTS
According to Franchi et al. in 2002 under a
high-fold optical microscope (x400), whether MMP-1
expression in tumor tissue was detected depended on
the basis of ≥ 10% positive cells after MMP-1 staining
of epithelial tissue20. IHC analysis of the tissue
samples showed that MMP-1 expression was higher
(≥10%) in 19 (63%) of 30 patients (Figure 1~3).
MMP-1 expression was higher in 4 of 13 patients with
stage I (n=7) or stage II (n=6) cancers compared to 15
of 17 patients with stage III (n=3) or stage IV (n=14)
cancers (Table 2). The difference between these
groups with early- and late-stage cancers was
statistically significant (p=0.002, Fisher’s exact test).
MMP-1 expression was higher in 8 (44%) of 18
patients with N0 disease, 4 (100%) of 4 with N1
disease, and 7 (88%) of 8 with N2 stage disease (Table
3). Differences in MMP-1 expression based on the
severity of lymph node involvement were statistically
significant (p=0.018, Fisher exact test). KruskalWallis H tests, post hoc comparisons, and Dunn’s tests
showed that MMP-1 expression intensity was high
Figure 2. Focal immunoreactivity for matrix metalloproteinase-1 (MMP-1) expression (40%) in the buccal
mucosa of a patient with squamous cell carcinoma
(T2N0M0G2).
in differentiation grade G2 and G3 patients and
significantly higher than that of patients with G1
disease (p=0.039 and 0.001, respectively) (Table 4).
DISCUSSION
Yamashita et al. pointed out that during the
multiple steps of tumor invasion and metastasis,
different types of MMPs have different functions.
For example, matrilysin and gelatinase break down
the basal membrane (type IV collagen), thus playing
a significant role in the metastasis of tumor cells.
Figure 1. Immunohistochemistry of matrix metalloproteinase-1 (MMP-1) in the tongue of a squamous cell
carcinoma patient (T4N2M0G3) revealing a high expression
level (60%). Most of the stained cells had a homogeneous
cytoplasmic pattern.
78
Figure 3. The buccal mucosa of a patient with squamous
cell carcinoma (T2N0M0G2) was almost completely negative
for matrix metalloproteinase-1 (MMP-1) immunohistochemical staining.
J Dent Sci 2008‧Vol 3‧No 2
MMP-1 expression in oral cancers
Table 2. MMP-1 expression associated with cancer stage
Cancer stage (I~IV)
MMP-1 negative expression
MMP-1 positive expression
Stages I and II
9 (69.2%)
4 (30.8%)
Stages III and IV
2 (11.8%)
15 (88.2%)
Fisher’s exact test (p=0.002).
Table 3. MMP-1 expression associated with lymph node staging
Lymph node metastasis stage (N0~N2)
MMP-1 negative expression
MMP-1 positive expression
10 (55.6%)
8 (44.4%)
1 (8.3%)
11 (91.7%)
N0
N1 and N2
Fisher’s exact test (p=0.018).
Table 4. MM P-1 expressional intensity associated with cancer differentiation in MMP-1-overexpressing patients
Cell Expression
post hoc comparisons
1 vs. 2
1 vs. 3
2 vs. 3
G
N
Mean
SD
Median
Minimum
Maximum
p- value
p- value
p- value
1
7
0.13
0.05
0.1
0.1
0.2
0.039
0.001
0.064
2
9
0.27
0.11
0.2
0.1
0.4
3
3
0.57
0.06
0.6
0.5
0.6
The overall test was performed by the Kruskal-Wallis H test.
Post hoc comparisons used Dunn’s test.
Collagenases mainly digest the ECM (type I collagen),
and thus participat in local invasion by tumor cells24.
Nelson et al.25 showed that interstitial cells in tumors
synthesize and secrete MMP-1, and this process is
especially prominent in fibroblasts at the periphery of
the tumor. Poletee et al.26 further suggested that in
head and neck tumors, MMP-1 is predominantly
expressed by fibroblasts and activated by interactions
between tumor tissue and interstitial cells. MMP-1
expression increases not only in tumor cells but also in
stromal cells (spindle cells, endothelial cells, and
monocytes). Eosinophils in the tumor stroma further
J Dent Sci 2008‧Vol 3‧No 2
regulate MMP-1 expression27. MMP-1 is believed to
be capable of remodeling tissue around tumor cells,
and interactions among the tumor, its stroma, and
inflammatory cells are thought to induce MMP-1 gene
expression. MMP-1 expression is also under the
control of many mediators, including interleukin-1α
and 1β, tumor necrosis factor-α, transforming growth
factor-α and -β, and keratinocyte growth factor28.
Ziober et al.29 found that MMP-1 secretion was
stimulated when epidermal growth factor (EGF) was
added to oral cancer cells from strain HSC-3, and that
EGF and type I collagen matrix can induce it. Turner
79
C.T. Chiu, S.Y. Lee, D.J. Wang, et al.
et al.30 demonstrated that EGF is an important
component of blood serum that can induce MMP
-1 secretion and degrade collagen. Using in situ
hybridization, Ziober et al.29 discovered that normal
mucosal epithelium rarely expresses MMP-1.
However, expression slightly increases in mucosal
dysplasia and carcinoma in situ, and it substantially
increases with highly invasive oral cancers.
The betel nut has been recognized as a carcinogen by the International Agency for Research on
Cancer31. Betel nut pieces for chewing in Taiwan
include the areca nut, the leaf of the Piper betel Linn,
the inflorescence, and mineral lime. The main
components of the betel nut include polyphenolic
compounds, alkaloids, tannins, thick fibers, lipids,
sugars, and iron. Arecoline is the major alkaloid in the
betel nut32. It has been proven that chewing betel nuts
is closely related with oral cancer. Betel chewing
alkalinizes the oral cavity. Under alkaline conditions,
some components of the betel nut release free radicals
such as superoxide radicals, hydroxyl anions, and
hydrogen peroxide (H2O2)33. Free radicals readily
cause cellular denaturation. Intracellular reactive
oxygen species (ROS) such as superoxide radicals,
H2O2, and nitric oxide (NO) play multiple roles in
general physiological and pathological conditions. An
excess of ROS will begin a chain reaction of free
radicals34, leading to the remodeling of the ECM and
changes in cell morphology. Nearly 150 patients with
newly diagnosed oral cancer are treated at Chi-Mei
medical center each year, and as many as 85%~95%
of them chew betel nuts on a regular basis. Our IHC
results revealed MMP-1 overexpression in tumor
epithelial cells among 19 (63%) of 30 patients, all of
whom had a long history of betel chewing. This result
demonstrates that MMP-1 plays a definitive role in the
development of oral cancer in Taiwan. Although the
mechanism of increased MMP-1 expression remains
unclear, it may be related to widespread betel nut
chewing in Taiwan. Long-term chewing of betel nuts
can produce persistent chronic inflammatory changes,
such as leukoplakia, submucosal fibrosis, and premalignant oral lesional changes. MMP-1 may also be
associated with wound healing. Therefore, MMP-1
expression in oral mucosal cells increases in people
who chew betel nuts for years because of persistent
chronic inflammation and tissue repair. This effect is
further potentiated in Taiwanese patients with oral
cancer and chronic buccal mucositis who regularly
chew betel nuts35. In our study, 60% of 30 patients
80
had cancer of the buccal mucosa or gum areas.
Additionally, it was noted that MMP-1 expression
increases when the disease progresses from early to
late stages. The MMP-1 expression intensity was
significantly higher in grade G2 and G3 than in grade
G1 of tumor cell differentiation. The findings revealed
that MMP-1 overexpression is related to the severity
of lymph node involvement and to the degree of
tumor cell dysplasia. These results confirm that MMP
-1 contributes to tumor development, proliferation,
lymph node metastasis, and tumor cell differentiation.
Constituents of betel nut can damage DNA and
cause cytotoxicity during differentiation of oral keratinocytes, inducing c-fos and c-jun expressions36, 37.
Persistent c-fos expression results in apoptosis. The
proto-oncogenes, c-fos and c-jun, are believed to
regulate many target genes and react rapidly to stimuli.
They may further induce MMP-1 expression, resulting
in ECM remodeling and alterations in cellular
morphology. c-fos is mainly induced by means of
the extracellular signal-regulated kinase (ERK)
transduction pathway. The mechanism of increasing
MMP-1 expression after exposure to betel quid extract
and arecoline may be related to regulation of the ERK
pathway since it regulates the MMP-1 gene promoter38.
Betel quid extract can activate the ERK and nuclear
factor-κB pathways in oral keratinocytes39. Arecoline can also activate the ERK pathway in oral
keratinocytes and oral epithelial cancer cells (KB)
cells40. Polyphenol compounds and alkaloids in betel
nuts may induce the formation of ROS and Nnitrosamines, resulting in tissue and cellular DNA
damage41,42. ROS can directly participate in tumor
initiation by inducing genetic toxicity. During betel
chewing, large amounts of ROS (e.g., H2O2) are
produced in cells of the oral cavity, which sustain
oxidative DNA damage43. An overabundance of ROS
may further activate free-radical cascades under the
influence of activator protein-1 (AP-1), remodeling of
the ECM, and alterations of cellular morphology.
MMP-1 expression, therefore, increases and accelerates injury to oral epithelial cells44.
MMP-1 expression is significantly higher among
Taiwanese patients with oral cancer who chew betel
nuts. As their disease progresses from early to late
stages, tumoral MMP-1 expression rises. Increased
MMP-1 expression was also related to the severity of
lymph node involvement and to the degree of cell
anaplasia. MMP-1 likely contributes to tumor
development, proliferation, lymph node metastasis,
J Dent Sci 2008‧Vol 3‧No 2
MMP-1 expression in oral cancers
and cell differentiation of oral cancer patients in
Taiwan. Therefore, inhibition of MMP-1 gene activity
may be a useful approach in future treatments of oral
cancer.
REFERENCES
1. Kumpawat K, Deb S, Ray S, Chatterjee A. Genotoxic effect
of raw betel-nut extract in relation to endogenous glutathione levels and its mechanism of action in mammalian cells.
Mutat Res, 538: 1-12, 2003.
2. Reichart PA, Philipsen HP. Betel chewer’s mucosa- a review.
J Oral Pathol Med, 27: 239-242, 1998.
3. Meghji S, Warnakulasuriya S. Oral submucous fibrosis: an
expert symposium. Oral Disease, 3: 276-297, 1997.
4. Cox SC, Walker DM. Oral submicous fibrosis: a review. Aust
Dent J, 41: 294-299, 1995.
5. Parsonnet J. Molecular mechanisms for inflammationpromoted carcinogenesis of cancer—the sixteenth international symposium of Sapporo cancer seminar. Cancer Res,
57: 3620-3624, 1997.
6. Deb S, Chatterjee A. Influence of buthionine sulfoximine and
reduced glutathione on arecoline-induced chromosomal
damage and sister chromatid exchange in mouse bone marrow
cells in vivo. Mutagenesis, 13: 243-248, 1998.
7. Nair J, Ohshima H, Friesen M, Croisy A, Bhide SV, Bartsch
H. Tobacco-specific and betel nut-specific N-nitroso compounds: occurrence in saliva and urine of betel quid chewers
and formation in vitro by nitrosation of betel quid.
Carcinogenesis, 6: 295-303, 1985.
8. Nagase H, Woessner JJ. Matrix metalloproteinases. J Biol
Chem, 274: 21491-21494, 1999.
9. Shapiro SD. Matrix metalloproteinase degradation of extracellular matrix: biological consequences. Curr Opin Cell
Biol, 10: 602-608, 1998.
10. Westermarck J, Kähäri VM. Regulation of matrix metalloproteinase expression in tumor invasion. FASEB J, 13:
781-792, 1999.
11. Kähäri VM, Saarialho KU. Matrix metalloproteinases and
their inhibitors in tumour growth and invasion. Ann Med, 31:
34-45, 1999.
12. Biswas C, Zhang Y, DeCastro R. The human tumour cellderived collagenase stimulatory factor (renamed EMMPRIN)
is a member of the immunoglobulin superfamily. Cancer Res,
55: 434-439, 1995.
13. Westermarck J, Li S, Jaakkola P. Activation of fibroblast
collagenase-1 expression by tumour cells of squamous cell
carcinomas is mediated by p38 mitogen-activated protein
kinase and c-Jun NH2-terminal kinase-2. Cancer Res, 60:
7156-7162, 2000.
14. Gomez DE, Alonso DF, Yoshiji H. Tissue inhibitors of
metallo- proteinases: structure, regulation and biological
functions. Eur J Cell Biol, 74: 111-115, 1997.
15. Murray GI, Duncan ME, O’Neil P, Melivin WT, Fothergill JE.
Matrix metalloproteinase-1 is associated with poor prognosis
in colorectal cancer. Nat Med, 2: 461-462, 1996.
J Dent Sci 2008‧Vol 3‧No 2
16. Zhu Y, Spitz MR, Lei L, Mills GB, Wu X. A single nucleotide
polymorphism in the matrix metalloproteinase-1 promoter
enhances lung cancer susceptibility. Cancer Res, 61: 78257829, 2001.
17. Murray GI, Duncan ME, O’Neil P, Melivin WT, Fothergill JE.
Matrix metalloproteinase-1 is associated with poor prognosis
in oseophageal cnacer. J Pathol, 185: 256-261, 1998.
18. Kurahara S, Shinohara M, Ikebe T. Expression of MMPs,
MT-MMP and TIMPs in squamous cell carcinoma of the oral
cavity: correlations with tumour invasion and metastasis.
Head Neck, 21: 627-638, 1999.
19. Sutinen M, Kainulainen T, Hurskainen T. Expression of
matrix metalloproteinases (MMP-1 and -2) and their
inhibitors (TIMP-1, -2 and -3) in oral lichen planus, dysplasia,
squamous cell carcinoma and lymph node metastasis. Br J
Cancer, 77: 2239-2245, 1998.
20. Franchi A, Santucci M, Masini E. Expression of matrix
metalloproteinase 1, matrix metalloproteinase 2, and matrix
metalloproteinase 9 in carcinoma of the head and neck.
Cancer, 95: 1902-1910, 2002.
21. O-Charoenrat P, Rhys-Evans P, Eccles S. Expression of
matrix metalloproteinases and their inhibitors correlates with
invasion and metastasis in squamous cell carcinoma of the
head and neck. Arch. Otolaryngol. Head Neck Surg, 127: 813820, 2001.
22. Nishioka Y, Kobayashi K, Sagae S, Ishioka S, Nishikawa A,
Matsushima M, Kanamori Y, Minaguchi T, Nakamura Y,
Tokino T, Kudo R. A single nucleotide polymorphism in
the matrix metalloproteinase-1 promoter in endometrial
carcinomas. Jpn J Cancer Res, 91: 612-615, 2000.
23. Tomasz K, Marcin F, Michal J, Marzena P, Teresa S, Tomasz
Z. Expression of collagenase-1 (MMP-1), collagenase-3
(MMP-13) and tissue inhibitor of matrix metalloproteinase-1
(TIMP-1) in laryngeal squamous cell carcinomas. Eur Arch
Otorhinolaryngo- logy, 260: 494-497, 2003.
24. Yamashita K, Mori M, Kataoka A, Inoue H, Sugimachi K.
The clinical significance of MMP-1 expression in oesophageal carcinoma. British Journal of Cancer, 84: 276-282,
2001.
25. Nelson AR, Fingleton B, Rothenberg ML, Matrisian LM.
Matrix metalloproteinases: biologic activity and clinical
implications. J Clin Oncol, 18: 1135-1139, 2000.
26. Polette M, Clavel C, Muller D, Abecassis J, Binninger I,
Birembaut P. Detection of mRNAs encoding collagenase I
and stromelysin 2 in carcinomas of the head and neck by in
situ hybridization. Invasion Metastasis, 11: 76-83, 1991.
27. Ono Y, Fujii M, Kameyama K, Otani Y, Sakurai Y, Kanzai J.
Expression of matrix metalloproteinase-1 mRNA related to
eosinophilia and interleukin-5 gene expression in head and
neck tumour tissue. Virchows Arch, 431: 305-310, 1997.
28. Wyatt CA, Coon CI, Gibson JJ, Brinckerhoff CE. Potential
for the 2G single nucleotide polymorphism in the promoter of
matrix metallo- proteinase to enhance gene expression in
normal stromal cells. Cancer Res, 62: 7200-7202, 2002.
29. Ziober BL, Turner MA, Palefsky JM, Banda MJ, Kramer RH.
Type I collagen degradation by invasive oral squamous cell
carcinoma. Oral oncology, 36: 365-372, 2000.
30. Turner MA, Darragh T, Palefsky JM. Epithelial–stromal
81
C.T. Chiu, S.Y. Lee, D.J. Wang, et al.
31.
32.
33.
34.
35.
36.
37.
82
interactions modulating penetration of matrigel membranes
by HPV 16- immortalized keratinocytes. Journal of Investigative Dermatology, 109: 619-625, 1997.
IARC. Betel-quid and areca-nut chewing and some areca-nut
related nitrosamines. IARC Monographs on the Evaluation of
Carcinogenic Risks to Humans, 85: 11-18, 2003.
Anluwalia HS, Ponnampalam JT. The socio-economic aspects
of betel-nut chewing. J Trop Med Hyg, 71: 48-50, 1968.
Nair UJ, Obe G, Friesen M, Goldberg MT, Bartsch H. Role of
lime in the generation of reactive oxygen species from
betel-quid ingredients. Environ Health Perspect, 98: 203-205,
1992.
Ito H, Duxbury M, Benoit E. Fibronectin-induced COX-2
mediates MMP-2 expression and invasiveness of rhabdomyosarcoma. Biochem Biophys Res Commun, 318:
594-600, 2004.
Chen YK, Huang HC, Lin LM, Lin CC. Primary oral
squamous cell carcinoma: an analysis of 703 cases in southern
Taiwan. Oral Oncol, 35: 173-179, 1999.
Yaar M, Gilani A, DiBenedetto PJ, Harkness DD, Gilchest
BA. Gene modulation accompanying differentiation of
normal versus malignant keratinocytes. Exp Cell Res, 206:
235-243, 1993.
Chang MC, Wu HL, Lee JJ, Lee PH, Chang HH, Hahn LJ,
Lin BR, Chen YJ, Jeng JH. The induction of prostaglandin E2
production, interleukin-6 production, cell cycle arrest, and
cytotoxicity in primary oral keratinocytes and KB cancer cells
by areca nut ingredients is differentially regulated by
MEK/ERK Activation. J Biol Chem, 279: 50676-50683,
2004.
38. Barchowsky A, Frleta D, Vincenti MP. Integration of the
NF-kappaB and mitogen-activated protein kinase/AP-1
pathways at the collagenase -1 promoter: divergence of IL-1
and TNF-dependent signal transduction in rabbit primary
synovial fibroblasts. Cytokine, 12: 1469-1479, 2000.
39. Lin SC, Lu SY, Lee SY, Lin CY, Chen CH, Chang KW. Areca
(betel) nut extract activates mitogen-activated protein kinases
and NF- kappaB in oral keratinocytes. Int J Cancer, 116:
526-535, 2005.
40. Chang MC, Wu HL, Lee JJ, Lee PH, Chang HH, Hahn LJ.
The induction of prostaglandin E2 production, interleukin-6
production, cell cycle arrest, and cytotoxicity in primary oral
keratinocytes and KB cancer cells by areca nut ingredients is
differentially regulated by MEK/ERK activation. J Biol Chem,
279: 50676-50683, 2004.
41. Hoffmann D, Brunnemann KD, Prokopczyk B. Tobaccospecific N-nitrosamines and Areca-derived N-nitrosamines:
chemistry, biochemistry, carcinogenicity and relevance to
humans. J Toxicol Environ Health, 41: 1-52, 1994.
42. Liu TY, Chen CL, Chi CW. Oxidative damage to DNA
induced by areca nut extract. Mutat Res, 367: 25-32, 1996.
43. Jacobs M D, Harrison SC. Structure of an IkappaBalpha /
NF-kappaB complex. Cell, 95: 749-758, 1998.
44. Brenneisen P, Wenk J, Klotz LO, Wlaschek M, Briviba K,
Krieg T, Sies H, Scharffetter-Kochanek K. Central role of
Ferrous/Ferric iron in the ultraviolet B irradiation-mediated
signaling pathway leading to increased interstitial collagenase
(matrix-degrading metalloprotease (MMP)-1) and stromelysin-1 (MMP-3) mRNA levels in cultured human dermal
fibroblasts. J Biol Chem, 273: 5279-5287, 1998.
J Dent Sci 2008‧Vol 3‧No 2