Vol.
8, 293–298, January 2002
An Orthotopic Nude Mouse Model of Oral Tongue Squamous
Cell Carcinoma
Jeffrey N. Myers,1 F. Christopher Holsinger,
Samar A. Jasser, B. Nebiyou Bekele, and
Isaiah J. Fidler
Departments of Head and Neck Surgery [J. N. M., F. C. H., S. A. J.],
Biostatistics [B. N. B.], and Cancer Biology [J. N. M., I. J. F.], The
University of Texas M.D. Anderson Cancer Center, Houston, Texas
77030-4009
ABSTRACT
Purpose: Despite advances in our understanding, pre-
vention, and treatment of head and neck squamous cell
carcinoma (SCC), the 5-year survival rates for patients re-
main low. This poor prognosis for head and neck SCC and
SCC of the oral tongue (SCCOT) in particular reflects a
limited understanding of the mechanisms of local and re-
gional metastasis, which accounts for a majority of deaths.
To analyze the molecular and cellular mechanisms of me-
tastasis, we have developed an orthotopic nude mouse model
of SCCOT.
Experimental Design: Nude mice were injected submu-
cosally in the tongue or subcutis with human squamous cell
carcinoma of the oral cavity cell lines Tu159, Tu167, and
MDA1986. The mice were necropsied and examined for the
presence of primary tumors, and regional and systemic
metastases.
Results: For all three of the squamous cell carcinoma of
the oral cavity cell lines, tumors developed more readily in the
orthotopic site, the tongue, than in the ectopic subcutis.
MDA1986 cells were highly tumorigenic, particularly at the
orthotopic site, with as few as 5 ⴛ 103 cells producing tumors
in all of the mice. In contrast, s.c. tumor formation required at
least 1 ⴛ 105 cells. The tumorigenicity observed between those
mice given submucosal inoculation and those mice given s.c.
inoculation (P < 0.0001). Regional metastases initially oc-
curred in <10% of mice. To generate tumor lines of increased
metastatic potential, regional metastases were isolated from
cervical lymph nodes after the development of orthotopic
tongue tumors. Serial passage of these lymph nodes resulted in
a cell line more metastatic than its parental line. When injected
Received 8/6/01; revised 10/5/01; accepted 10/8/01.
The costs of publication of this article were defrayed in part by the
payment of page charges. This article must therefore be hereby marked
advertisement in accordance with 18 U.S.C. Section 1734 solely to
indicate this fact.
1
To whom requests for reprints should be addressed, at Department of
Head and Neck Surgery, The University of Texas M.D. Anderson
Cancer Center, 1515 Holcombe Boulevard, Box 441, Houston, Texas,
77030-4009. Phone: (713) 792-6920; Fax: (713) 794-4662; E-mail:
jmyers@mdanderson.org.
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Clinical Cancer Research 293
into the tongues of mice, these cells metastasized to regional
lymph nodes in 30% of mice and to the lungs in 20%.
Conclusions: In this orthotopic murine model, oral
tongue cancer recapitulates the behavior of human SCCOT,
allowing for detailed studies of its biology and therapy.
Introduction
HNSCC2 remains a significant public health problem (1).
Cancers of the oral cavity and pharynx alone account for
363,000 new cases worldwide and almost 200,000 deaths an-
nually (2). In 2001, these tumors are predicted to account for
30,100 new cancers, representing 3% of all cancers in the
United States, equal in incidence to leukemia and greater than
all of the endocrine tumors. In particular, SCCOT is among the
most common tumors of the head and neck (3). Despite ad-
vances in our understanding, prevention, and treatment, the
5-year survival rates for patients with SCCOT have not im-
proved in the past 25 years and remain ⬃50%. This high
mortality is considerably worse than rates for breast, cervical, or
colorectal cancer (3). This poor prognosis for SCCOT may
reflect a limited understanding of the mechanisms of local and
regional metastasis, which accounts for a majority of deaths.
Regional metastasis strongly correlates with tumor recur-
rence and mortality (4 – 8). Extension of cervical metastases
outside the lymph node capsule (extracapsular spread) is an
even worse clinical indicator (9, 10). In a recent retrospective
review of ⬎250 patients from 1980 –1995 with SCCOT, overall
and disease-specific survival rates for patients with extracapsu-
lar spread were 30% and 50%, respectively (11). Distant me-
tastasis developed in 24.4% of patients with regional metastasis
but in only 3.3% of patients without neck disease.
Despite these abundant data, little is known regarding the
tumor biology of regional metastases of SCCOT. Furthermore,
there is no reliable animal model for study. Therefore, we have
developed an orthotopic model of SCCOT metastases by inject-
ing cell lines from human oral cavity SCC into the tongue of
nude mice. In this model, local tumor growth, and regional and
distant metastases demonstrated a histopathological similarity to
SCCOT primary tumors from patients. Finally, these tumors
recapitulate the regional and distant metastatic patterns seen in
patients with SCCOT.
MATERIALS AND METHODS
Cell Lines
Three lines of human SCCOC (Tu167, Tu159, and
MDA1986), established from freshly resected human tumors
2
The abbreviations used are: HNSCC, head and neck squamous cell
carcinoma; SCC, squamous cell carcinoma; SCCOT, squamous cell
carcinoma of the oral tongue; GFP, green fluorescent protein; SCCOC,
squamous cell carcinoma of the oral cavity.
294 Mouse Model of SCCOT
(12) were obtained from the laboratory of Dr. Gary L. Clayman,
The University of Texas M.D. Anderson Cancer Center. B16-
BL6 melanoma cell lines were used to determine the feasibility
of the orthotopic injection procedure and have been described
previously (13–15).
GFP Transfection of SCCOC Cell Lines and Selection for
GFP Expression
PEGFP-C1 plasmid (Clontech, Palo Alto, CA) was trans-
fected into the cell lines by the CaPO4 method using 2 M CaCl2
and 2 ⫻ Hanks’ balanced, buffered solution (Calbiochem, San
Diego, CA). The cells were plated onto 100-mm dishes at a
density of 1 ⫻ 106 cells/dish. The cultures were incubated after
transfection at 37°C in an incubator under 5% CO2 for 16 h;
cultures were then washed and refed with fresh DMEM (high
glucose) supplemented with 10% fetal bovine serum, L-gluta-
mine, nonessential amino acids, penicillin-streptomycin, sodium
pyruvate, and vitamins. To select clones with resistance to
neomycin, 400 ␮g/ml G418 (Life Technologies, Inc., Gaithers-
burg, MD) in DMEM was added starting 48 h after transfection.
Two weeks later, the G418-resistant clones were expanded, and
2 ⫻ 106 cells/cell line were analyzed on a flow cytometer at the
highest excitation and emission wavelengths to view GFP. The
top 5% of fluorescent cells were isolated and cultivated at 37°C
in a 5% CO2 incubator.
Animal Care and Injections
Male athymic nude C57BL/6 mice, 6 –10 weeks of age, were
purchased from the National Cancer Institute (Bethesda, MD) and
housed in a specific pathogen-free animal facility. The animals
were fed irradiated mouse chow and autoclaved reverse osmosis
treated water. All of the animal procedures were performed in
accordance with a protocol approved by the Institutional Animal
Care and Usage Committee. To determine whether these human
tumor cell lines would grow better when implanted at an orthotopic
site, i.e., a tissue bed most closely resembling the primary site of
origin, we injected SCCOC in both the tongues and flanks of the
experimental animals. All of the mice were anesthetized with
sodium pentobarbital (50 mg/kg body weight). Cells from all four
of the tumor lines (B16-BL6, Tu167, Tu159, and MDA1986) were
then prepared in 50 ␮l of Hanks’ balanced, buffered solution in the
following dilutions: 1 ⫻ 103, 5 ⫻ 104, 1 ⫻ 104, 5 ⫻ 105, 1 ⫻ 106,
and 5 ⫻ 106 cells. A group of five mice were used for each
concentration.
Subcutaneous Flank Injection Technique
In these groups, each mouse underwent s.c. injection of
cells (suspended in a volume of 50 ␮l) directly into the flank
using a 1-ml tuberculin syringe (Hamilton Co., Reno, NV) with
a 30-gauge hypodermic needle. Mice were then examined every
other day for tumor development. When present, tumors were
measured using calipers in cephalad-to-caudad and left-to-right
dimensions. The maximal tumor diameter was recorded.
Orthotopic Sublingual Injection Technique
In these groups, each mouse then underwent submucosal
injection cells (suspended in a volume of 50 ␮l) directly into
anterior tongue using a 1-ml tuberculin syringe (Hamilton Co.)
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with a 30-gauge hypodermic needle. Mice were then examined
every other day for the development of tongue tumors and
weight loss. The mice were killed by CO2 inhalation when they
had lost ⬎ 25% of their preinjection body weight.
In Vivo Selection of Cell Lines for Increased
Metastatic Potential
Using a similar technique, a GFP-expressing Tu167 SC-
COC cell line, Tu167GFP was injected into the tongues of nude
mice. Regional metastases developed, and the mice were killed
using CO2 inhalation. A standard apron-flap incision was made
in the neck of the experimental animal. Lymph nodes were
harvested by aseptic techniques, dissociated mechanically using
a wire mesh sieve, and placed into culture. Cells were grown in
DMEM with 20% fetal bovine serum, sodium pyruvate, nones-
sential amino acids, L-glutamine, and a 2-fold vitamin (Life
Technologies, Inc.). To inhibit fibroblast overgrowth of cultures
and allow selection of tumor cells, 300 ␮g/ml G418 were added
to the cultures. These tumor lines were harvested within three to
five in vitro passages and reinjected into the tongues of athymic
nude mice. These cells were then injected submucosally into the
tongue of several nude mice. The mice were killed after they had
lost 25–30% of their body weight. The cervical lymph nodes
were harvested by sterile dissection techniques and placed in
antibiotic containing medium. The lymph nodes were dissoci-
ated, and tumor cells were cultured for several weeks (as a
monolayer) in a 5% C02 incubator at 37°C until colonies of
epithelial cells could be identified. To select for the growth of
cells that had been transfected previously with GFP and the
neomycin phosphotransferase gene, the cells were then grown in
medium containing G418 (400 ␮g/ml). The neomycin-resistant
cells were expanded and designated as the Tu167G-LN1 cell
line. Tu167G-LN1 cells were then injected into the tongue of a
group of nude mice (n ⫽ 10), which were killed when they lost
⬎25% of initial body weight. Cervical lymph nodes from 2 mice
were sterilely resected and grown in tissue culture to generate
the Tu167LN2 cell line. The cervical lymph nodes of the re-
maining 8 mice and the lungs of all 10 of the mice were fixed
in formalin, embedded in paraffin, and evaluated by light mi-
croscopy after staining with H&E.
Necropsy and Tissue Preparation
After the mice were killed, the tongues, cervical lymph
nodes, and viscera (lungs, liver, and brain) were removed and
placed in formalin solution overnight. Each specimen was em-
bedded in paraffin and sectioned. Sections were stained with
H&E and evaluated by light microscopy for the presence of
regional or distant metastases.
Microscopy
A Leica (model MZ FLIII) fluorescence dissecting ste-
reomicroscope was used to visualize fluorescent metastases. The
microscope was equipped with a 100-W, mercury vapor lamp
power source and fitted with a GFP filter set. Images were
processed using Image Pro Plus (version 4.0; Media Cybernet-
ics, L.P., Silver Spring, MD) and Adobe PhotoShop (version
5.5; Adobe Systems Inc., San Jose, CA).

Statistical Analyses
Tumorigenicity of MDA1986 SCCOC. For this pilot
study, standard maximum likelihood based methods were not
applicable. Exact logistic regression was used to assess both the
tumorigenicity of SCCOC in mice and the effects of orthotopic
(submucosal) versus ectopic (s.c.) tumor development. Four to
five mice were inoculated at each level/dilution combination (56
mice total). The regression model included terms for the dilution
effect (ranging from 1 ⫻ 103 to 5 ⫻ 106 cells) and for the
inoculation location (submucosal versus s.c.). The Ps for the
location effect were evaluated against an ␣ significance level of
0.05.
Selection of Human SCCOC Cell Line Tu167 for
Greater Metastatic Potential. Fisher’s exact test and cross-
tabulation was used to compare the rates of regional and lung
metastasis in 45 mice inoculated with the TU167 cell line versus
10 mice injected with Tu167GLN1 cell line. The p value for this
test was evaluated against an ␣ significance level of 0.05.
All of the statistical computations were performed on a
Dell 600 mHz PC using the SAS statistical system and S-plus
(Cary, NC).
RESULTS
Human SCCOC Cell Lines Form Tumors when In-
jected Submucosally into the Tongues of Nude Mice. The
highly metastatic B16-BL6 melanoma cells, implanted into
the tongues of the syngeneic C57 BL/6 mice, formed tumors in
the tongues of all of the injected mice and yielded high rates of
regional metastases (Fig. 1, A and B), demonstrating the feasi-
bility of this orthotopic (submucosal lingual) injection tech-
nique. Next, we injected three different human SCCOC cell
lines (MDA1986, Tu159, and Tu167) into the tongues of nude
mice. The injection of 5 ⫻ 106 cells produced tongue tumors in
all of the injected mice, regardless of the cell line used. The
orthotopic SCCOC tumors resulting from submucosal injection
of Tu167 (Fig. 1C) were very similar in histological appearance
to that of primary human SCCOT specimens (Fig. 1D), dem-
onstrating keratin pearls and intercellular bridging.
Tumorigenicity in Orthotopic and Ectopic Organs.
For the three human SCCOC cell lines, a spectrum of tumori-
genicity was observed related to tumor cell line, concentration,
and site of injection. These trends are described below.
SCCOC Tu159. Tongue tumors developed in all of the
mice injected with 5 ⫻ 104, 1 ⫻ 105, 5 ⫻ 105, 1 ⫻ 106, and 5 ⫻
106 Tu159 cells. s.c. tumors developed in none of the animals
inoculated with 5 ⫻ 104 cells and in only one of five animals
and three of five animals inoculated with Tu159 at 1 ⫻ 105 and
5 ⫻ 105 cells, respectively. All of the mice injected 1 ⫻ 106 and
5 ⫻ 106 with Tu159 cells developed s.c. tumors. These data
show that the Tu159 cells are more tumorigenic at the orthotopic
site than the ectopic s.c. tissue.
SCCOC Tu167. In the mice injected with Tu167, tongue
tumors were found in all of the animals injected submucosally
with at least 1 ⫻ 105 cells. However, only one of five mice
developed s.c. tumors when injected with 1 ⫻ 105, 5 ⫻ 105, or
1 ⫻ 106 cells, indicating at least a 50-fold difference in the
minimal tumorigenic dose between the tongue and subcutis
(Table 1).
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Clinical Cancer Research 295
SCCOC MDA1986. The MDA1986 cell line was highly
tumorigenic, particularly at the orthotopic site, with as few as
5 ⫻ 103 cells producing tumors in all of the mice. In contrast,
s.c. tumor formation required an injection of at least 1 ⫻ 105
cells. The results of the tumorigenicity study of the MDA1986
cell line are summarized in Table 1. Furthermore, in the exact
logistic regression model the difference in tumorigenicity ob-
served between those mice given submucosal inoculation and
those mice given s.c. inoculation was statistically significant
(P ⬍ 0.0001).
The Metastatic Pathways of Human SCCOT Are
Recapitulated in the Orthotopic Nude Mouse Model of
SCCOT. For mice injected sublingually with the Tu167 tumor
cells, only 4% developed lymph node metastasis, and none of
these animals had detectable pulmonary metastases. Two meth-
ods were used to identify cervical and distant metastasis. Im-
munohistochemical staining with an anticytokeratin antibody
(data not shown) was performed to confirm the epidermoid
character of each metastasis.
Selection of SCCOC Cell Lines for Greater Metastatic
Potential. This initially low rate of regional metastases for
Tu167 provided us with a model system for characterizing the
biological processes and specific molecules critical for the de-
velopment of cervical and systemic metastases. Our objective
was to generate tumor lines of increased metastatic potential by
identifying regional metastases within the small, cervical lymph
nodes of mice after the development of orthotopic tongue
tumors.
At first, it was quite difficult to identify these uncommon
metastases, in part because the unique location of the tongue
leads to rapid weight loss, necessitating early sacrifice. To
identify microscopic regional metastases earlier, the Tu167 cell
line was transfected with GFP, and the resultant cell line, Tu167
tumor cells expressing GFP, was orthotopically implanted into
the tongue of the nude mouse. Visible lingual tumors developed,
and the mice were sacrificed after ⬎25% preinjection weight
loss. At necropsy, fluorescence stereomicroscopy was used to
identify metastatic GFP-transfected tumor cells. This method
readily identified both primary tongue tumors (Fig. 1E) and
submandibular metastasis of GFP-expressing Tu159 (Fig. 1F).
Both of tumors were confirmed histologically (Fig. 1G).
Metastatic tumor cells from lymph nodes containing a
fluorescent tumor were harvested and grown in culture. The
resulting tumor cell line, Tu167G-LN1, was expanded and re-
injected into the tongues of 10 mice. With this new cell line,
cervical (Fig. 1H) and distant metastases (Fig. 1I) were seen
with greater frequency (see Table 2). Using Fisher’s exact test,
there was a statistically significant difference in regional and
lung metastases rates between Tu167 and Tu167G-LN1 (P ⫽
0.0004).
DISCUSSION
Treatment failure in HNSCC results primarily from re-
gional and distant metastasis. To study the mechanism of
metastasis in SCCOT, we have developed a reliable, ortho-
topic nude mouse model. First, we verified the feasibility of
sublingual injection and the development of regional metas-
tases using the pigmented B16-BL6 melanoma cell line in
296 Mouse Model of SCCOT

Fig. 1 A and B, regional metastasis from orthotopic sublingual implantation of B16-BL6 melanoma. The tongues of C57BL/6 black mice were
inoculated with syngeneic B16-BL6 melanoma cells, and mice were killed after 14 days and necropsy performed. An obvious tumor is seen in the
tongue. Bilateral metastases are easily identified by melanotic pigmentation within the cervical lymph nodes. C and D, comparative histopathology
showing the resemblance between orthotopic oral tongue cancer in the nude mouse and human SCCOT. Photomicrographs of (C) the Tu167 human
SCCOC cell line grown orthotopically in the tongue of nude mouse and (D) a human SCCOT tumor. E–G, visualization of orthotopic tongue tumor
and regional metastasis in situ with GFP. Tu159GFP was inoculated submucosally in the tongue of each mouse; the mouse was killed, and the tumor
was visualized under a dissecting fluorescence microscope. The primary tumor in the tongue (E) is readily apparent. An area of fluorescence, noted
in the submandibular region (F), was resected, revealing metastatic tumor on microscopic examination (G; H&E). H, regional metastasis of the
orthotopic TU167G-LN1 SCCOC. Photomicrograph of a cervical lymph node from a nude mouse in which the Tu167G-LN1 line was grown
orthotopically, showing a subcapsular metastatic tumor (H&E). I, distant pulmonary metastasis from the orthotopic TU167G-LN1 SCCOC.
Photomicrograph of a lung from a nude mouse in which the Tu167G-LN1 line was grown orthotopically, revealing distant metastatic tumor (H&E).

syngeneic mice. Next, three different human SCC oral cavity
tumor lines were then injected submucosally into the tongues
of nude mice: Tu167, derived from a primary tumor of the
floor of mouth; Tu159, from a primary SCCOT; and
MDA1986, cultured from a cervical metastasis of human
SCCOT. All of the tumor lines were more tumorigenic in the
orthotopic site than in the subcutis. This nude mouse model
of oral cavity cancer substantiates the orthotopic principle
that tumor cells grow better in their tissue of origin than in an
ectopic (s.c.) site.
The orthotopic nude mouse model also provides an invalu-
able method for in vivo selection and generation of tumor cell
lines of greater regional and distant metastatic potential. Ini-
tially, only 4.4% of mice injected with Tu167 developed cervi-
cal lymph node metastases. After serial passages, the more
metastatic cell line Tu167G-LN1 was identified. When ortho-
topically reimplanted, Tu167G-LN1 cells produced a 30% rate
of regional lymphatic metastasis. Pulmonary metastasis did not
develop in mice injected with Tu167 cells, whereas 20% of the
mice injected with Tu167G-LN1 had metastases in the lungs.

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Table 1 Tumorigenicity of MDA1986 SCCOC in ectopic versus
orthotopic tissues
Groups of 6 –10-week old nude mice were inoculated s.c. along the
flank or submucosally in the tongue with the indicated numbers of cells
of the human oral cavity SCC cell line, MDA1986. Mice were then
observed for tumor formation.
% mice with
MDA1986 (no. % mice with s.c. tongue tumors
cells injected) tumors (ectopic) (orthotopic)
1 ⫻ 103 0 100%
5 ⫻ 104 0 100%
1 ⫻ 104 40% 100%
5 ⫻ 105 100% 100%
1 ⫻ 106 100% 100%
5 ⫻ 106 100% 100%
We are continuing this in vivo passage for several more cycles
to select for cell lines of greater metastatic potential. By devel-
oping SCCOC cell lines with low and high metastatic propen-
sity, it may be possible to identify the molecular basis of
metastasis in SCCOT.
An orthotopic nude mouse model to investigate the cellular
and molecular mechanisms of metastasis in human neoplasia
was first described by Fidler et al. (15, 16) and Killion et al.
(17). Other groups using human solid tumors from a variety of
organ sites have subsequently documented its significance (18,
19). The orthotopic implantation of tumor cells restores the
correct tumor-host interactions, which do not occur when tu-
mors are implanted in ectopic s.c. sites (15). Orthotopic models
can properly evaluate the metastatic propensity of human tu-
mors and select cell lines of varying metastatic potentials. In
addition, an accurate experimental animal model is necessary to
evaluate the efficacy of novel therapies (17, 19).
Previous orthotopic models of oral cancer have met with
limited success. The first report of orthotopic growth of human
oral cavity tumor lines in nude mice was published by Fitch et
al. (20), who aspirated cells from fresh human tumors growing
s.c. in nude mice and injected them into the tongues of nude
mice. These studies showed an equal tumorigenicity in the oral
cavity and the subcutis. Certain SCCOT cell lines that grew
better in the ectopic subcutis of the nude mouse may have
accounted for these results.
An oral cavity model, developed by Dinesman et al. (21),
relied on transcutaneous injection via a submandibular route into
the tissue deep to the mylohyoid muscle beneath the floor of mouth.
Whereas only 5% of these tumors metastasized to regional lymph
nodes, 40% of animals developed pulmonary metastases. Whereas
recent reports using this model have highlighted its potential utility
in quantifying local invasiveness into muscle and bone (22, 23), the
injection of cells transcutaneously into the deep neck muscles
differs from submucosal injection. Tumor seeding and spillage may
predispose these animals to develop hematogenous pulmonary
metastasis, contradicting an orthotopic approach. This may account
for the higher incidence of pulmonary metastasis compared with
regional lymphatic metastasis.
An orthotopic model of cutaneous SCC metastases was
developed by Chen et al. (24), using a transformed skin kerati-
nocyte line that was injected s.c. in syngeneic mice. After
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Clinical Cancer Research 297
Table 2 Selection of human SCCOC cell line Tu167 for greater
metastatic potential
Groups of 6 –10-week-old nude mice were inoculated submuco-
sally in the tongue with 1 ⫻ 106 cells of the SCC cell line Tu167 or
Tu167GLN1. The latter cell line was derived from in vivo passage of
Tu167 in the orthotopic SCCOC tumor model by placement of a cervical
lymph node in culture. Mice were then observed for tumor formation
and subjected to necropsy, and the number of mice with cervical and/or
pulmonary metastases was tabulated.
No. mice with regional No. mice with lung
Cell line metastasis metastasis
Tu167 2/45 0/45
TU167GLN1 3/10 2/10
passage in vivo before harvesting metastatic variants from the
lymph nodes, the resultant cell lines expressed higher levels of
chemokine growth-regulated oncogene-␣, interleukin-8, and nu-
clear factor ␬B than the parental cells (25). Our mouse model
offers an ideal and analogous experimental system to study the
molecular basis of metastasis in mucosal HNSCC.
Orthotopic nude mouse models provide invaluable insights
into metastasis, both at the molecular and cellular level, and are
a vital first step in identifying safe and effective new therapies.
However, nude mouse models have limitations. Data acquired
should be additionally substantiated with complementary stud-
ies with immunocompetent models and analysis of archival
human, oral tumor specimens. All of these studies provide
critical information necessary for preclinical assessment of new
drugs to fight head and neck cancer.
In summary, we have developed a model of SCCOT to
study regional lymph node metastases and distant visceral
metastases. This model should facilitate in vivo studies of the
systemic cellular and molecular mechanisms of tumorigenic-
ity, growth, and metastasis in SCCOT. Whereas no animal
model can correlate directly with the human metastatic proc-
ess, this orthotopic animal model will play an important role
in the evaluation of novel therapies for the treatment of
HNSCC.
ACKNOWLEDGMENTS
We thank Michael Wilson, Carol Oborn, and Joseph Douglas for
their technical assistance.
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An Orthotopic Nude Mouse Model of Oral Tongue Squamous
Cell Carcinoma
Jeffrey N. Myers, F. Christopher Holsinger, Samar A. Jasser, et al.

Clin Cancer Res 2002;8:293-298.

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