ORIGINAL ARTICLE

CORRELATION OF THE EXPRESSION OF HUMAN KALLIKREIN-

RELATED PEPTIDASES 4 AND 7 WITH THE PROGNOSIS IN
ORAL SQUAMOUS CELL CARCINOMA
Hong Zhao, MD,1,2 Ying Dong, PhD,3 Jingjing Quan, MSc,1 Robert Smith, PhD,1 Alfred Lam, MD,1
Stephen Weinstein, MD,4 Judith Clements, PhD,3 Newell W. Johnson, PhD,1 Jin Gao, PhD1
1
Grifth Institute for Health and Medical Research, School of Dentistry and Oral Health, and School of Medicine,
Grifth University, Queensland 4222, Australia. E-mail: j.gao@grifth.edu.au
2
Tianjin Medical University Eye Hospital, Tianjin 300070, Peoples Republic of China
3
Hormone Dependent Cancer Program, Institute of Health and Biomedical Innovation, Queensland University of Technology,
Kelvin Grove, Queensland 4059, Australia
4
Pathology Queensland, Gold Coast Hospital, Queensland 4215, Australia

Accepted 22 April 2010

Published online 27 July 2010 in Wiley Online Library (wileyonlinelibrary.com). DOI: 10.1002/hed.21496

est improvements in survival rates over the past sev-

Abstract: Background. Very few articles have been written
about the expression of kallikreins (KLK4 and KLK7) in oral
cancers. Therefore, the purpose of this study was to examine
and report on their prognostic potential.
Methods. Eighty archival blocks of primary oral cancers
were sectioned and stained for KLK4 and KLK7 by immunohis-
tochemistry. The percentage and the intensity of malignant ke-
ratinocyte staining were correlated with patient survival using
Cox regression analysis.
Results. Both kallikreins were expressed strongly in the
majority of tumor cells in 68 of 80 cases: these were mostly
moderately or poorly differentiated neoplasms. Staining was
particularly intense at the inltrating front. Patients with intense
staining had signicantly shorter overall survival (p < .05).
Conclusion. This is the rst observation on the patient sur-
vival inuenced by kallikrein expression in oral carcinoma. The
ndings are consistent with those for carcinomas at other sites,
in particular the prostate and ovary. KLK4 and/or KLK7 immu-
nohistochemistry seems to have diagnostic and prognostic
potential in this disease. V C 2010 Wiley Periodicals, Inc.
Head Neck 33: 566572, 2011
Keywords: oral squamous cell carcinoma; kallikrein serine
proteases; immunohistochemistry; prognosis
Oral and pharyngeal squamous cell carcinoma repre-
sents the sixth most common cancer worldwide.1 If
cancer of the larynx, which shares risk factors with
oral and pharyngeal cancer, is added, these head and
neck malignancies constitute an extremely serious
public health problem. Furthermore, in many coun-
tries, oral squamous cell carcinoma (OSCC) itself
accounts for the major proportion of these malignan-
cies, with signicant impact on the patients quality
of life and poor prognosis. There have been only mod-
Correspondence to: J. Gao
Contract grant sponsor: Grifth University, Queensland, Australia.
V
C 2010 Wiley Periodicals, Inc.
eral decadesand then in only the most advanced
treatment centers.2,3
The progression of epithelial malignancies requires
the involvement of proteolytic enzymes directly or
indirectly in many processes, including proliferation,
destruction of basement membranes and invasion of
connective tissues, chemotaxis, and angiogenesis.4,5
Although many of the complex proteolytic networks
involved are well known, others, such as those belong-
ing to the human tissue kallikrein (KLK)-related pepti-
dase family of serine proteases, have not been
extensively studied in OSCC.6
The KLK family of genes consists of 15 members,
many of which are highly expressed in a number of
cancers compared to their normal parent tissues.7,8
KLK4 is thought to be involved in the degradation of
extracellular matrix proteins, thus promoting cancer
cell invasion.9,10 Overexpression of KLK4 has been
reported to be associated with poor prognosis in cancers
of the prostate and ovary.1113 It has also been sug-
gested that KLK4 might be involved in the establish-
ment of bone metastasis of prostate cancer, because it is
highly expressed in these lesions.14 Further evidence
for involvement in mineralized tissue function and the
oral cavity comes from its expression by ameloblasts at
the early and late bell stages of enamel organ transition
and mineralization during tooth development.15
KLK7 was initially characterized as an enzyme
implicated in the degradation of intercellular cohesive
structures in the stratum corneum of stratied squa-
mous epithelia, preceding desquamation in skin.16 It
catalyzes the degradation of desmosomes in the outer-
most layer of skin and permits cell shedding to take
place at the skin surface.1719 Overexpression of KLK7
in tumor cells has been reported to signicantly
enhance the invasive potential in intracranial malig-
nancies and ovarian cancer cells.2023

566 Kallikreins 4 and 7 in Oral Cancer HEAD & NECKDOI 10.1002/hed April 2011
 Table 1. Clinicopathological characteristics of 80 patients with oral squamous cell carcinoma (OSCC).

Sex T N M Histodifferentiation

Site/oral cavity M F Age, mean 0, 1, 2, 3, 4 0, 1, 2, 3 0, 1 W M* P*

Floor of mouth 4 5 59 1, 3, 2, 1, 2 2, 3, 1, 0 8, 1 4 4 1
Palate 6 2 52 0, 0, 4, 1, 3 0, 3, 1, 0 8, 0 2 6 0
Tongue 20 17 63 0, 21, 9, 5, 2 0, 1, 2, 2 36, 1 12 19 6
Other intraoral areas 14 12 49 1, 9, 7, 6, 3 0, 5, 5, 3 23, 3 4 19 3
Total cases 44 36 2, 33, 22, 13, 10 2, 12, 9, 5 75, 5 22 (27%) 48 (60%) 10 (13%)
Abbreviations: M, male; F, female; W, well-differentiated; M, moderately differentiated; P, poorly differentiated.
*p value (p < .05).

Thus, KLK4 and KLK7 could contribute to the
degradation of extracellular matrices in OSCC tis-
sues, promoting invasion of neoplastic cells locally
and facilitating metastasis to regional lymph nodes.
The present study describes the expression of KLK4
and KLK7 in primary human OSCC by immunohisto-
chemistry and, for the rst time, the association of
that expression with patient survival times. Our
results justify further evaluation of the potential of
KLK4 and KLK7 enzymes as prognostic markers for
this disease.
MATERIALS AND METHODS
Archival Blocks and Clinical Data. A total of 80
archival parafn blocks were retrieved from Gold
Coast Hospital Queensland Health Pathology Serv-
ices, consisting of 80 patients, 36 women and 44 men,
with ages ranging from 35 to 94 years (mean age, 62
years), with research ethics clearance from the rele-
vant committees of the Gold Coast Hospital, Queens-
land Health, and Grifth University. These were from
patients with OSCC between September 1997 and De-
cember 2003 who underwent surgical resection of the
primary neoplasm as initial treatment. Patient demo-
graphics, the sites of the primary lesion, TNM status,
and histodifferentiation, obtained from the original
patient medical records, are given in Table 1. Patients
were subsequently treated by various combinations of
further surgery, radiotherapy, and chemotherapy, but
these variables were not included in the present
analysis.
body, rabbit polyclonal anti-peptide KLK4 or anti-
KLK7 (Figure 1), as described previously,14,21 was
applied at a concentration of 2 lg/mL and incubated at
4
C overnight. For detection of primary antibody bind-
ing, an antirabbit EnVision Systems secondary anti-
body (DAKO, Botany, Australia) was used at room
temperature for 15 minutes, followed by 3 washes with
PBS, incubation with diaminobenzadine Chromogen
(DAB) solution (DAKO) for 4 minutes, and nally coun-
terstained with Mayers hematoxylin. Sections were
then dehydrated and mounted with DPX (BDH Labo-
ratory, Poole, England). Tissue sections were visualized
under light microscopy (Leitz Laborlux S, Germany)
and photographed using a Nikon OXM1200 digital
camera with the Act-1 program.
For negative controls, primary antibodies were
replaced by normal rabbit sera. Sections from a prostate
cancer known to stain positively for both KLK4 and
KLK7 were used as positive controls in each analysis.
All experiments were duplicated.
Assessment Criteria for Immunohistochemistry
Staining Intensity and Cell Counts. The intensity of
immunohistochemical staining for KLK4 and KLK7
was scored on a scale of to . Staining dis-
tributed anywhere within the cell as few but unequiv-
ocal ne yellow-brown grains represented weak
staining, and recorded as ; moderate staining was
dened as easily visible yellow-brown grains, and
strong staining as intense dark yellow-brown grains,

Preparation of Tissue Sections and Immunohisto-

chemistry. Archival tissue blocks, originally forma-
lin-xed and parafn embedded, were sectioned at 5
lm and placed on aminopropyltriethoxysilane pre-
coated slides using standard techniques. Sections
were deparafnized by xylene and rehydrated in
graded ethanol and distilled water. For antigen re-
trieval, sections were incubated in a BORG decloaker FIGURE 1. Schematic structures show protein organisation of
KLK4 and KLK7. Indicated are the pre-region (Pre), pro-region
(pH 9.0; Biocare Medical, Concord, Australia) and
(Pro) and mature region of the enzymes; the catalytic triad, his-
heated in a pressure cooker at 125
C for 4 minutes. tidine (His), aspartate (Asp) and serine (Ser). Arrows indicate
Sections were equilibrated in phosphate buffer solution the localisation of the peptides used to raise the anti-KLK4-N
(PBS) for 10 minutes and incubated in 1% bovine se- and anti-KLK7-N antibodies employed in this study. [Color gure
rum albumin in PBS to block endogenous peroxidase can be viewed in the online issue, which is available at
for 30 minutes at room temperature. The primary anti- wileyonlinelibrary.com.]

Kallikreins 4 and 7 in Oral Cancer HEAD & NECKDOI 10.1002/hed April 2011 567
FIGURE 2. (A) Immunostaining with kallikrein (KLK)4 antibody reveals light deposits of enzyme in several layers of normal oral epithe-
lium (OE) and more strongly within malignant keratinocytes in the main tumor mass (oral squamous cell carcinoma [OSCC]; bar
100 lm). (B) Higher magnication reveals distribution of KLK4 in both cytoplasm and nuclei (arrow) of malignant cells (bar 25 lm).
(C) KLK4 is strongly expressed in cancer cells invading muscle (arrows; bar 50 lm). (D) KLK7 expression is negative or very weak
in normal oral epithelium (OE), but strong within the tumor mass (OSCC, bar 150 lm). (E) At higher magnication KLK7 is seen
predominantly in the cytoplasm (arrows, bar 25 lm). (F) KLK7 is also strongly expressed in malignant keratinocytes (arrows) inltrat-
ing nerves (bar 50 lm). [Color gure can be viewed in the online issue, which is available at wileyonlinelibrary.com.]

respectively scored as and . Absence of
staining was scored as negative. Slide interpretation
was recorded by 3 observers and the results were dis-
cussed and pooled.
All sections were examined by 2 observers inde-
pendently using a graticule eyepiece micrometer
(Olympus, 24 OC-M, 10/10 SQ) under 400 magnica-
tion. The total number of cells stained positively for
KLK4 or KLK7 and nonstained cells were counted
from 10 adjacent small squares (25 lm2) of the grati-
cule randomly. This was duplicated in a second ran-
domly selected microscopic eld. Mean and SDs of cell

568 Kallikreins 4 and 7 in Oral Cancer HEAD & NECKDOI 10.1002/hed April 2011
 Table 2. Number of cases (%) of positively stained primary oral squamous cell carcinoma (OSCC) by site and histodifferentiation.

KLK4 positive KLK7 positive

Site in oral cavity W M* P* W M* P*

Floor of mouth 50 75 100 25 75 100

Palate 50 67 N/A 50 83 N/A
Tongue 45 78 100 50 89 100
Other intraoral area 50 84 100 50 73 100
Total cases 10/22 (43%) 38/48 (79%) 10/10 (100%) 10/22 (43%) 39/48 (77%) 10/10 (100%)
Abbreviations: KLK, kallikrein; W, well-differentiated; M, moderately differentiated; P, poorly differentiated; N/A, not applicable.
The intensity of immunostaining for KLK4 and KLK7 was set at or above, and >70% of positively stained cells only were included.
*p value (p < .05).

counts were determined: these were then grouped at
<10%, 30%, and >70%, respectively.
Statistical Analysis. Data were analyzed by SPSS
(version 17). The ndings between the staining inten-
sity, or the percentage of positively stained cells for
KLK4 or KLK7 expression, and clinicopathologic
parameters, in particular histodifferentiation and
patient survival, were analyzed using the 2-sided chi-
square and by Spearman correlation tests. Disease-
free survival (DFS) was dened as the interval
between surgery and the rst documented evidence of
recurrent disease in a locoregional disease and/or dis-
tant sites. Overall survival was dened as the interval
between surgery and death from the disease. DFS and
overall survival curves were calculated using the
KaplanMeier method and the log-rank test was used
to compare survival curves. Univariate and multivari-
ate relative risks were calculated using the Cox pro-
portional hazard regression. Probability of < .05 was
considered statistically signicant.
RESULTS
Clinicopathologic Parameters. Table 1 shows that
27% (n 22) of the cancers were well-differentiated,
60% (n 48) were moderately differentiated, and
13% (n 10) were poorly differentiated. According to
the TNM classication, 41% (n 33) were stage I, 29%
(n 22) were stage II, 16% (n 13) were stage III, and
14% (n 10) were stage IV. The majority of oral cancers
were from the tongue (37 of 80 cases): of these, 12 were
well-differentiated, 19 were moderately differentiated,
and 6 were poorly differentiated. At the time of diagno-
sis, 32% of 80 patients (n 26) had clinically positive
neck nodes; 5 patients had lung metastasis and died 6
months later.
Staining Patterns of Kallikrein-4 and Kallikrein-7 in
Oral Squamous Cell Carcinoma. KLK4 was weakly
expressed in normal oral epithelium adjacent to the
primary tumor. Staining was spread over all layers of
the epithelium, most strongly in basal cells (Figure
2A). All invasive cancer cells were stained positively
with moderate to strong intensity. Most cells showed
both cytoplasmic and nuclear staining (Figure 2B).
Signicantly, the strongest staining intensity was
seen in the deepest invading cells surrounding muscle
bundles and nerves (Figure 2C). Staining intensity of
to was seen in 43% of well-differentiated
cases, over 79% of moderately differentiated cases,
and all poorly differentiated cases (Table 2). Cell
counts revealed that over 50% of total cancer cells
were positively stained in most cases. There were
statistically signicant differences in KLK4 expres-
sion between well-differentiated, and moderately dif-
ferentiated or poorly differentiated cancers (p < .05).
KLK7, on the other hand, did not stain normal
oral epithelium or was noted as very weak staining.
Within the neoplasm, staining was predominantly
cytoplasmic (Figure 2D). Like KLK4, staining for
KLK7 was also strong in neoplastic cells inltrating
muscles and nerves (Figure 2E and F). The propor-
tion of cases with staining intensity at or above
was similar to that of KLK 4% to 43% in well-differ-
entiated, 77% in moderately differentiated, and 100%

Table 3. Correlation of patients 5-year survival with immunostaining intensity for KLK4 and KLK7 within
oral squamous cell carcinoma (OSCC) cases (%).

Floor of mouth Palate Tongue Other intraoral area

Site in oral cavity KLK4 KLK7 KLK4 KLK7 KLK4 KLK7 KLK4 KLK7

Follow-up
Alive 67 50 50 50 54 46 67 78
Died* 100 100 100 100 88 93 70 70
Unknown 100 100 50 100 50 83 50 50
Abbreviation: KLK, kallikrein.
*p value (p < .05).

Kallikreins 4 and 7 in Oral Cancer HEAD & NECKDOI 10.1002/hed April 2011 569
FIGURE 3. (A) Cox regression analysis of patient survival according to the intensity of kallikrein (KLK) staining. There were no differ-
ences in staining intensity, case by case, between KLK4 and KLK7 expression. There is markedly poorer survival in cases with higher
intensity (p < .05). Cum Survival cumulative survival (%). "Censored cases are cases for which death is not recorded, but for
whom tracking has ended. (B) The percentage of both KLK4 and KLK7 positive staining is well correlated with patient survival by Cox
regression analysis. The higher percentage, the poorer survival is noted (p < .05). [Color gure can be viewed in the online issue,
which is available at wileyonlinelibrary.com.]

in the poorly differentiated lesions (Table 2). Simi-
larly, cell counts showed that the highest percentage
of positively stained cells (70%, >90% of total cells)
was observed in poorly differentiated cancers: this,
however, was not signicantly different between well-
differentiated and moderately differentiated cancers.
Overall, however, the staining intensity was not
signicantly different between KLK4 and KLK7, but
the histodifferentiation was correlated with the stain-
ing intensity throughout most specimens in which
KLK4 and KLK7 were detected. For example, for the
poorer differentiated cancers, the higher staining in-
tensity was noted in over 70% of the cases (Table 2).
It was found that the staining intensity of both KLK4
and KLK7 correlated with OSCC patients survival
status (Table 3). No staining was seen in the negative
controls.
Survival Analysis. All patients were followed up for
at least 5 years, the average 5-year survival rate
being 43%. Average survival time was 31.4 months.
Recurrence of the primary cancer was noted in 25
patients, occurring between 9 and 51 months. Distant
metastasis was recorded in 5 patients. Cancer differ-
entiation had no signicant effect on both DFS and
overall survival (data not shown). Patients diagnosed
with poor TNM grading of the lesions had a signi-
cantly shorter life span (p < .05), such as those with
positive nodal status and distant metastasis at diag-
nosis. However, the overall TNM stage of the cancer
had no signicant effect on the overall survival in
this small study population.
In contrast, Cox regression analysis reveals a
striking difference in survival according to both the
staining intensity and the percentage of positively
stained KLK4 and KLK7 cases (Figure 3A and B).
DISCUSSION
KLKs have been reported to play important roles in
physiological processes from embryonic development
to tissue remodeling, and in the spread of malignant
neoplasms by assisting degradation of basement
membranes and of the extracellular matrix of connec-
tive tissues.7,9,10 Overexpression of KLKs in ovarian,
breast, and prostatic carcinomas has suggested their
potential as prognostic markers,14,2123 but their role
in the development and progression of head and neck
cancers has not been intensively investigated.6,24
It was reported that mutations in the human
KLK4 gene could lead to the autosomal recessive dis-
ease, amelogenesis imperfecta, a group of disorders of
tooth enamel development due to hypoplasia and/or
hypomineralization.25 KLK7 was shown to promote
pancreatic carcinoma invasion by shedding E-cad-
herin26 and urinary type plasminogen activator recep-
tor and decrease adhesion to vitronectin.27 This is
further reected in OSCC cells,6 as when transfected
with urinary type plasminogen activator and its
receptor urinary type plasminogen activator receptor,
SCC25 cells showed marked changes in KLK
expression with 2.8, 5.3, 4.0, and 3.5-fold increases of
KLK5, KLK7, KLK8, and KLK10, respectively. Fur-
thermore, their studies revealed that these KLKs
were highly regulated in both murine and human
OSCC tumors.
Our present study showed that both KLK4 and
KLK7 were detected in a cohort of 80 OSCC tissue
samples. They were expressed most strongly in the
deeply invading cells, implying their potential roles in

570 Kallikreins 4 and 7 in Oral Cancer HEAD & NECKDOI 10.1002/hed April 2011
local tumor invasion. However, the tissue distribution
patterns between KLK4 and KLK7 are different.
KLK4 was found throughout all layers of the normal
oral epithelium, whereas KLK7 was noted in outer
layer cells but not in the basement membrane. In
addition, KLK4 was localized in both the cytoplasm
and nuclei, whereas staining of KLK7 was mostly
accumulated in the cytoplasm, not in the nuclei. The
staining characteristic of both KLK4 and KLK7 in
cancer cells was consistent with previous studies.11,13
The reasons for these differences are not yet clear.
KLK4 was initially found in the highly mineralized
enamel matrix secreted by functional ameloblasts dif-
ferentiated from the inner-enamel epithelium at the
secretary phase during tooth development.15 Accumu-
lating evidence suggests that certain tissue KLKs
may be part of enzymatic cascades critically activated
in some cancers. We have demonstrated its involve-
ment in bone metastasis of prostate cancer, particu-
larly in mediating cross-talk between cancer and bone
cells.14 A study of 147 ovarian cancer tissues reported
that high expression of KLK4 was associated with
more aggressive lesions and with poor prognosis.13
We also reported that KLK4 expression increased in
prostate cancer tissues, and the combined effect of
KLK4 and KLK3 could induce an epithelial mesen-
chymal transition in prostate cancer cells, making the
tumor cells increasingly undifferentiated and more
aggressive.26,28,29
KLK7 is primarily characterized for its specic
function in degrading the intercellular cohesive struc-
tures in supercial layers of the skin.1618 We now
know that expression levels are signicantly higher
in poorly differentiated ovarian cancers in which
KLK7 is thought to have value as a prognostic indica-
tor.2123 It is also suggested that KLK7 may be
involved in the invasion and metastasis of human cer-
vical squamous cell carcinoma.30 Despite little infor-
mation available on the mechanisms of KLK7 in
cancers progression, we recently demonstrated that
the a5b1-integrin pathway was likely to be involved
in promoting the peritoneal dissemination and re-
invasion of ovarian carcinoma.31
Early studies of KLKs in oral cancer showed
promising outcomes with the report of increased
expression of several members of the KLK-serine pro-
tease family in OSCC cells.6 Our present study pro-
vides additional data to indicate the potential value of
KLK4 and KLK7 in determining prognosis of OSCC.
The distinct functions of those genes need to be fur-
ther investigated in OSCC cell lines by using siRNA
approaches, to test whether the knockdown of KLK4
or KLK7 gene expression affects the invasion or met-
astatic abilities of OSCC cells in vitro. This may lead
to consideration of specic targeted therapies via in-
hibition of kallikrein protease-driven mechanisms in
OSCC, as part of the growing armamentarium with
potential for specic, individualized management of
patients with head and neck cancer.
Kallikreins 4 and 7 in Oral Cancer
REFERENCES
1. Warnakulasuriya S. Global epidemiology of oral and oropharyn-
geal cancer. Oral Oncol 2009;45:309316.
2. Johnson N. Global epidemiology of oral cancer. In: Shah J, John-
son N, Batsakis J, Dunitz M, Oral Cancer. New York: Thieme
Medical; 2003. pp 332.
3. Jemal A, Siegel R, Ward E, Hao Y, Xu J, Thun MJ. Cancer sta-
tistics, 2009. CA Cancer J Clin 2009;59:225249.
4. Levental KR, Yu H, Kass L, et al. Matrix crosslinking forces tu-
mor progression by enhancing integrin signaling. Cell 2009;139:
891906.
5. Shibue T, Weinberg RA. Integrin beta1-focal adhesion kinase sig-
naling directs the proliferation of metastatic cancer cells disse-
minated in the lungs. Proc Natl Acad Sci U S A 2009;106:10290
10295.
6. Pettus JR, Johnson JJ, Shi Z, et al. Multiple kallikrein (KLK 5,
7, 8, and 10) expression in squamous cell carcinoma of the oral
cavity. Histol Histopathol 2009;24:197207.
7. Yousef GM, Diamandis EP. Tissue kallikreins: new players in nor-
mal and abnormal cell growth? Thromb Haemost 2003;90:716.
8. Lawrence MG, Lai J, Clements JA. Kallikreins on steroids:
structure, function, and hormonal regulation of prostate-specic
antigen and the extended kallikrein locus. Endocr Rev 2010.
[Epub ahead of print].
9. Debela M, Beaufort N, Magdolen V, et al. Structures and speci-
city of the human kallikrein-related peptidases KLK 4, 5, 6,
and 7. Biol Chem 2008;389:623632.
10. Paliouras M, Borgono C, Diamandis EP. Human tissue kallik-
reins: the cancer biomarker family. Cancer Lett 2007;249:6179.
11. Xi Z, Klokk TI, Korkmaz K, et al. Kallikrein 4 is a predomi-
nantly nuclear protein and is overexpressed in prostate cancer.
Cancer Res 2004;64:23652370.
12. Obiezu CV, Scorilas A, Katsaros D, et al. Higher human kallik-
rein gene 4 (KLK4) expression indicates poor prognosis of ovar-
ian cancer patients. Clin Cancer Res 2001;7:23802386.
13. Dong Y, Kaushal A, Bui L, et al. Human kallikrein 4 (KLK4) is
highly expressed in serous ovarian carcinomas. Clin Cancer Res
2001;7:23632371.
14. Gao J, Collard RL, Bui L, Herington AC, Nicol DL, Clements
JA. Kallikrein 4 is a potential mediator of cellular interactions
between cancer cells and osteoblasts in metastatic prostate can-
cer. Prostate 2007;67:348360.
15. Hu JC, Zhang C, Sun X, et al. Characterization of the mouse
and human PRSS17 genes, their relationship to other serine pro-
teases, and the expression of PRSS17 in developing mouse inci-
sors. Gene 2000;251:18.
16. Simon M, Jonca N, Guerrin M, et al. Rened characterization of
corneodesmosin proteolysis during terminal differentiation of
human epidermis and its relationship to desquamation. J Biol
Chem 2001;276:2029220299.
17. Debela M, Hess P, Magdolen V, et al. Chymotryptic specicity deter-
minants in the 1.0 A structure of the zinc-inhibited human tissue
kallikrein 7. Proc Natl Acad Sci U S A 2007;104:1608616091.
18. Komatsu N, Takata M, Otsuki N, et al. Expression and localization
of tissue kallikrein mRNAs in human epidermis and appendages.
J Invest Dermatol 2003;121:542549.
19. Caubet C, Jonca N, Brattsand M, et al. Degradation of corneo-
desmosome proteins by two serine proteases of the kallikrein
family, SCTE/KLK5/hK5 and SCCE/KLK7/hK7. J Invest Derma-
tol 2004;122:12351244.
20. Prezas P, Scorilas A, Yfanti C, et al. The role of human tissue
kallikreins 7 and 8 in intracranial malignancies. Biol Chem
2006;387:16071612.
21. Dong Y, Kaushal A, Brattsand M, Nicklin J, Clements JA. Dif-
ferential splicing of KLK5 and KLK7 in epithelial ovarian cancer
produces novel variants with potential as cancer biomarkers.
Clin Cancer Res 2003;9:17101720.
22. Prezas P, Arlt MJ, Viktorov P, et al. Overexpression of the
human tissue kallikrein genes KLK4, 5, 6, and 7 increases the
malignant phenotype of ovarian cancer cells. Biol Chem
2006;387:807811.
23. Kyriakopoulou LG, Yousef GM, Scorilas A, et al. Prognostic
value of quantitatively assessed KLK7 expression in ovarian
cancer. Clin Biochem 2003;36:135143.
24. Choi S, Myers JN. Molecular pathogenesis of oral squamous cell
carcinoma: implications for therapy. J Dent Res 2008;87:1432.
25. Hart PS, Hart TC, Michalec MD, et al. Mutation in kallikrein 4
causes autosomal recessive hypomaturation amelogenesis imper-
fecta. J Med Genet 2004;41:545549.
HEAD & NECKDOI 10.1002/hed April 2011 571
26. Johnson SK, Ramani VC, Hennings L, Haun RS. Kallikrein 7
enhances pancreatic cancer cell invasion by shedding E-cad-
herin. Cancer 2007;109:18111820.
27. Ramani VC, Haun RS. Expression of kallikrein 7 diminishes
pancreatic cancer cell adhesion to vitronectin and enhances uro-
kinase-type plasminogen activator receptor shedding. Pancreas
2008;37:399404.
28. Veveris-Lowe TL, Lawrence MG, Collard RL, et al. Kallikrein 4
(hK4) and prostate-specic antigen (PSA) are associated with
the loss of E-cadherin and an epithelial-mesenchymal transition
(EMT)-like effect in prostate cancer cells. Endocr Relat Cancer
2005;12:631643.
572 Kallikreins 4 and 7 in Oral Cancer
29. Klucky B, Mueller R, Vogt I, et al. Kallikrein 6 induces E-cad-
herin shedding and promotes cell proliferation, migration, and
invasion. Cancer Res 2007;67:81988206.
30. Santin AD, Cane S, Bellone S, et al. The serine protease stratum
corneum chymotryptic enzyme (kallikrein 7) is highly overex-
pressed in squamous cervical cancer cells. Gynecol Oncol
2004;94:283288.
31. Dong Y, Tan OL, Loessner D, et al. Kallikrein-related pepti-
dase 7 promotes multicellular aggregation via the alph(5)-
beta(1) integrin pathway and paclitaxel chemoresistance in
serous epithelial ovarian carcinoma. Cancer Res 2010;70:2624
2633.
HEAD & NECKDOI 10.1002/hed April 2011