The Prostate 43:263–271 (2000)

Use of Nude Mouse Xenograft Models in

Prostate Cancer Research
Wytske M. van Weerden* and Johannes C. Romijn
Department of Experimental Urology, Josephine Nefkens Institute, Erasmus University
Rotterdam, The Netherlands

BACKGROUND. Our understanding of the mechanisms of (progressive) growth of prostatic

cancer has been largely obtained through the study of experimental animal models. To be able
to validate new concepts, representative model systems of human origin that mimic the
clinical process of the disease in patients are essential. Unfortunately, the limited number of
human prostate tumor models has considerably hampered research.
METHODS. Various research groups have put much effort in the development of human
prostate tumor xenograft models, and large numbers of clinical prostate tumors were hetero-
transplanted in immune-deficient host animals. This huge effort has resulted in a number of
tumor lines which are reviewed here.
RESULTS. Up to now, approximately 25 xenograft models of human prostate cancer have
been established and reported in the literature. The available xenografts seem to represent the
various stages of clinical prostate cancer, such as early progression and transition from an-
drogen-dependent to androgen-independent growth. In addition, recent efforts are concen-
trating on the establishment of in vitro cell lines from these xenografts as well as on the
development of (bone) metastatic variants.
CONCLUSIONS. Xenograft models are important for elucidating regulatory pathways of
tumor growth and progression and are indispensible for testing of new treatment modalities.
Prostate 43:263–271, 2000. © 2000 Wiley-Liss, Inc.

KEY WORDS: xenograft models; human prostate cancer; immunodeficient mice

INTRODUCTION in culture or in nude mice. Also, animal models of

With the increasing life expectancy of men, prostat-
ic cancer has become a major medical problem. Al-
though (advanced) prostate cancer is in most cases
initially treatable by antiandrogen therapy, the indefi-
nite escape from hormonal control and consequently
the relapse in tumor growth is one of the main clinical
problems [1]. In addition, metastatic spread of pros-
tate cancer towards bone is the main cause of morbid-
ity among prostate cancer patients [2]. To improve the
present treatment modalities, new therapeutic targets,
related to these issues, have to be identified. Repre-
sentative animal models are needed to develop and
test new concepts of properties and prostate cancer
growth regulation.
Research on human prostate cancer has for many
years been hampered by the very limited availability
of experimental model systems. The most important
reason for the lack of representative models is the very
poor growth potential of human prostate tumor tissue
prostatic cancer are hardly available since other mam-
mal species (e.g., laboratory animals), in contrast to
men, do not, or only rather poorly, develop prostatic
cancer spontaneously upon aging [3]. The spontane-
ous occurrence of prostate carcinoma in aging dogs
and ACI/Seg and Lobund-Wistar rats has been used
in model sytems. In addition, different hormonally
and/or chemically induced rodent model systems
have been developed successfully. In recent years, a
number of transgenic mouse models of prostate cancer
have been established [4]. Although these model sys-
Grant sponsor: Dutch Cancer Society; Grant number: EUR97-1479;
Grant sponsor: CapCURE.
*Correspondence to: W.M. van Weerden, Ph.D., Experimental Urol-
ogy, BE 355A, Josephine Nefkens Institute, Erasmus University,
P.O. Box 1738, 3000DR Rotterdam, The Netherlands.
E-mail: vanweerden@uro.fgg.eur.nl
Received 7 December 1999; Accepted 20 December 1999

© 2000 Wiley-Liss, Inc.

264 van Weerden et al.
TABLE I. Transplantation of Human Prostate Tumor Tissue in Athymic Nude Mice*
Mouse
strain Year PC LN
BALB/c 1988 2 3
1989 12 4 0
1990 7 3
Total 21 (60%) 10 (29%)
NMRI 1990 3 0
1991 4 2
Total 7 (44%) 2 (13%)
*PC, primary prostate carcinoma; LN, lymph node metastasis; TURP, transurethral resection of the prostate; DM, distant metastasis
(scrotal skin metastasis).
tems all contribute to our understanding of the fun-
damental aspects of prostate cancer biology, the cor-
relation to the clinical situation remains to be
determined. Representative model systems of human
origin are essential for validation of the concepts de-
veloped from these animal models.
MATERIALS AND METHODS
The introduction of the mouse mutant nude as a
host for heterotransplantation of human cancer tissue
opened a new era of cancer research. The mouse mu-
tant nude was first reported by Isaacson and Cattan-
ach in 1962 [5]. As a consequence of the absence of a
functional thymus, the mouse mutant nude had a de-
ficient cell-mediated immune response [6]. Although
humoral antibody formation is only slightly impaired
and the activity of natural killer (NK) cells is actually
increased in these immunosuppressed animals, sub-
cutaneous grafting of human malignant tissue readily
resulted in tumor development transplantation [7,8].
Unfortunately, this proved to be problematic for all
tumor types; however, many of the efforts to develop
human prostate xenograft models appeared to be un-
successful.
The Rotterdam Experience
It was only in 1977 that the first transplantable hu-
man prostate xenograft, PC-82, was established. The
Experimental Urology Research Group of the Depart-
ment of Urology of Erasmus University Rotterdam,
headed by Prof. Dr. Schröder, started in the 1970s with
attempts to develop human prostate cancer xenograft
models, using their own breeding colony of athymic
nude mice of BALB/c background. Human prostate
cancer tissue was directly transported from the clinic
into the laboratory. After confirmation of tumor tissue
by the pathologist, small tumor fragments were trans-
planted subcutaneously in both shoulders of male
TURP DM Total Take
3 0 8 0
0 16 0
1 0 11 1
4 (11%) 0 35 (100%) 1 (3%)
1 0 4 1
5 1 12 5
6 (38%) 1 (5%) 16 (100%) 6 (38%)
and/or female nude mice. In 1977, these efforts re-
sulted in the first transplantable human prostatic car-
cinoma reported, the androgen-responsive PC-82 tu-
mor model [9]. In subsequent years, over 150
specimens of prostatic carcinomas were transplanted,
which finally resulted in another two permanently
growing, androgen-independent xenografts, PC-133
and PC-135, established in 1981 and 1982, respectively.
The very low success rate of approximately 5% was a
figure that was also recognized by other investigators
in the field at that time. In 1981, Hoehn et al. were able
to establish a second androgen-responsive human
prostate xenograft, PC-EW, from a metastatic lesion in
lymph nodes [10]. This tumor model was studied in
Rotterdam extensively, and showed many character-
istics of the PC-82 tumor [11]. Unfortunately, this tu-
mor was found in later years to be infected with the
mouse hepatitis virus and, for that reason, was re-
moved from our panel of available tumor models.
We continued our efforts to establish additional xe-
nograft models with a relatively small number of cases
per year without much success. In 1990–1992, we were
able to add an additional seven new xenograft models
to our existing panel in a relatively short period of
time [12]. To explain this dramatically enhanced suc-
cess rate, we compared the various factors that were
thought to interfere with the take rate of heterotrans-
planted tumor tissue. First of all, a comparison of the
origin of patient material was made between the dif-
ferent years (Table I). Tumor material was obtained
from primary prostate tumors of untreated patients
undergoing radical prostatectomy, positive lymph
nodes of untreated patients, transurethral resections of
the prostate (TURP) of hormonally-treated, relapsed
patients, and finally, occasionally, a distant metastatic
lesion. The number of transplantations varied consid-
erably over the years, but was never quite large and
was certainly not highly different in those successful
years. Also, the origin of the tumor material did not
show a large variation, although the number of trans-
 Xenograft Models in Prostate Cancer Research 265

TABLE II. Human Prostate Cancer Xenograft Models*

Tumor Nu/nu mouse Reference

model Origin Matrigel AD/AI AR PSA no.

CWR22 Prostate ?+M + Mutant + 26

LAPC-3 Prostate SCID + M − Wt + 30
LAPC-4 Lymph node SCID + M + Wt + 30
LAPC-9 Bone SCID + M + Wt + 31
LuCaP 23.1 Lymph node BALB/c + N/A + 33
LuCaP 23.8 Lymph node BALB/c + N/A + 33
LuCaP 23.12 Liver BALB/c + N/A + 33
MDA Pca-31 Liver BALB/c N/A + + 29
MDA Pca40 Liver BALB/c N/A − − 29
MDA Pca-43 Adrenal BALB/c N/A + + 29
MDA Pca-44 Skin BALB/c N/A − − 29
PC-82 Prostate BALB/c + Wt + 9
PC-133 Bone BALB/c − − −
PC-135 Prostate BALB/c − − −
PC-295 Lymph node NMRI + Wt + 12
PC-310 Prostate NMRI + Wt + 12
PC-324 TURP NMRI − − − 12
PC-329 Prostate NMRI + Wt + 12
PC-339 TURP NMRI − − − 12
PC-346 TURP NMRI + Wt + 12
PC-346I NMRI − Mutant +
PC-346B NMRI + Wt +
PC-346BI NMRI − Wt +
PC-374 Skin NMRI − Wt + 12

*M, Matrigel; TURP, transurethral resection of the prostate; AD, androgen dependence (+); AI, androgen independence (−); AR, human
androgen receptor; PSA, prostate-specific antigen; Wt, wild-type receptor; N/A, not available. PC-346 was derived from an nonpro-
gressed TURP patient; three sublines have been derived from this xenograft. PC-374 is still sensitive to androgen treatment.

planted lymph node metastases decreased in the years
1990 and 1991, while the number of TURP specimens
increased from 11% to 38% of the total number of
transplanted patient material in these years. In our
opinion, these relatively small variations cannot ac-
count for the increase in success rate from 3% to 38%.
In these years, the use of reconstituted basement
membrane (Matrigel) was introduced for its growth-
promoting effect when coinjected with tumor cells
[13,14]. In contrast to other research groups, we de-
cided not to use this method for our xenograft devel-
opment project. We felt that the effects of Matrigel
treatment on the original patient material could be
quite considerable, especially with regard to differen-
tiation, as reported by others [15,16]. Thus, the patient
tumor tissue was simply cut into small fragments and
transplanted subcutaneously in both shoulders of athy-
mic nude mice without any pretreatment. Also, in sub-
sequent propagation of xenografts we never made use of
Matrigel, and all tumors were serially passaged by sub-
cutaneous transplantation of small tumor fragments.
The host animals are the second source of factors
that might influence the take rate of xenotransplanted
tissue. The animals used were either intact males or
testosterone- or dihydrotestosterone-supplemented
male or female mice. Neither of these hormonal con-
ditions seemed, however, to have any effect on the
take rate of tumors. What might have been an impor-
tant factor in this success was the change from the
BALB/c athymic nude mouse as host animals for het-
erotransplantation to the NMRI athymic nude. The
NMRI mouse strain was developed in the Naval Ma-
rine Research Institute (Bethesda, MD), and was intro-
duced in 1960 into Europe [17]. In Hamburg, Wagner
et al. reported on the increased succes rate of hetero-
transplantations in these animals [18]. The use of
NMRI nude mice was found to be rather successful in
our hands, indeed, but no data are available on com-
parisons of xenotransplantations in both mouse
strains to really prove this point statistically. More-
over, once the new xenograft was established in NMRI
nude mice, we could serially passage the xenograft in
BALB/c nude mice as well, suggesting that if the
NMRI mouse was important at all, it was only during
the first passage in the mouse. It is known that the
NMRI nude mouse is relatively sensitive to (trans-
266 van Weerden et al.
plantation-induced) sarcomas, and other tumors (lym-
phomas) are relatively frequently formed in NMRI
mice in response to transplantation [19,20]. It is, there-
fore, essential to check the histology of the serially
transplanted xenograft with every mouse passage.
Unfortunately, and mainly due to our success, we
were not able to continue our efforts to develop new
xenograft models for practical as well as financial rea-
sons. Meanwhile other groups, mainly in the USA,
also started to increase their efforts to establish new
prostate xenograft models. This has resulted in a con-
siderable number of interesting models, of which the
important aspects will be highlighted here (Table II).
RESULTS
The Rotterdam PC-Models
In Table II, some important characteristics of the
whole panel of human prostate xenograft models are
summarized [12]. The present panel comprises 10 xe-
nograft models of 9 different patients. The PC-346 and
PC-346B tumors are derived from different TURP
specimens of the same patient. Growth characteristics
as well as tissue-specific markers are differently ex-
pressed between PC-346 and PC-346B, which have
been established in exactly the same way but on sepa-
rate animals. Their differences reflect the obvious
heterogeneity of the original patient material. Unfor-
tunately, we recently lost the strictly androgen-
dependent PC-329 model. This tumor model had
much of the characteristics well-known for the PC-82
tumor model. However, it was an extremely slow-
growing tumor (tumor doubling time of more than 20
days), which developed only after 4–5 months follow-
ing transplantation.
All xenograft models have been propagated into
intact male mice from their establishment onwards.
The passage numbers of the different xenograft lines
range from 25–70, depending on their growth rate.
Interestingly, most initial characteristics have been
preserved rather well, and no significant changes have
been observed in their growth behavior as well as
their androgen responsiveness. We have three models,
PC-82, PC-295, and PC-310, that are strictly dependent
on androgens for their development and growth: these
tumors do not develop in castrated males, and cas-
tration of tumor-bearing animals results in tumor re-
gression without tumor relapse [21–23]. PC-295 and
PC-310 are interesting model systems, since neuroen-
docrine differentiation is induced concomitantly with
the regression in tumor volume upon androgen deple-
tion [23,24]. The PC-346 model, originating from an
untreated patient, harbors androgen-independent fea-
tures, but is androgen-responsive. Tumors do develop
in castrated males with a prolonged lag phase and
reduced growth rate, but growth of this tumor is
clearly stimulated by the addition of androgen. Cas-
tration of tumor-bearing animals consistently results
in a variable response, ranging from tumor regression
to tumor retardation to continued tumor. As in pros-
tate cancer patients, all regressed or retarded PC-346
tumors will ultimately continue to grow. Meanwhile,
several sublines have been developed from both PC-
346 and PC-346B, which now constitute an interesting
subpanel, with differences in androgen responsive-
ness representing the variations in progressive pros-
tate cancers.
The PC-374 xenograft, like PC-346, is androgen-
responsive, but not androgen-dependent: tumors de-
velop in castrated male mice and continue to grow
after androgen ablation, but at a slower growth rate.
Also from this xenograft, a truly androgen-indepen-
dent subline could be established. Finally, the PC-133,
PC-135, PC-324, and PC-339 xenografts are the truly
androgen-independent representatives of prostatic
cancer. These tumors develop identically in intact or
castrated males, and their growth rate does not change
upon testosterone depletion or supplementation.
In all xenograft models except for PC-133, high lev-
els, or in case of the androgen-independent tumors,
low levels of hAR protein were detected. All xeno-
grafts but one have the wild-type hAR, as analyzed by
single-stranded conformation polymorphism (SSCP)
of exons 2–8 of the hAR gene. Interestingly, in one
androgen-independent subline of the PC-346 tumor, a
mutation was detected in the ligand-binding domain
of hAR (T877A), which was identical to the mutation
known to be present in the LNCaP human prostate
cancer cell line. As is the case in the LNCaP cell line,
estrogens indeed stimulated growth of this AR-
mutated PC-346 subline, whereas estrogen-induced
growth was not observed in the wild-type PC-346 xe-
nograft (results not shown).
The CWR Series of Xenografts
In 1993, Pretlow et al. [25] first reported on their
efforts to develop new human prostate tumor xeno-
grafts using Matrigel. The CWR series of tumors was
serially propagated as cell suspensions mixed with
Matrigel. The first reported tumor, CWR22, originated
from a primary tumor and showed secretion of PSA
and was said to be androgen-responsive [26]. Later
reports showed androgen response after castration of
tumor-bearing mice. Relapse of tumor growth in the
absence of androgen is observed in approximately 25–
50% of the animals within 3–10 months of castration.
Interestingly, a reduction in circulating PSA levels
 Xenograft Models in Prostate Cancer Research
preceded this relapse in tumor growth. Relapsed an-
drogen-independent xenografts (CWR22R) could be
established from CWR22 [27]. Recently, the character-
ization of hAR of the CWR22 tumor was reported,
revealing that this tumor harbored a missense muta-
tion in the ligand-binding domain of the hAR gene
(H874Y) [28]. The mutant AR was transcriptionally
active upon stimulation by T and DHT. However, the
adrenal androgen dehydroepiandrosterone (DHEA),
as well as estradiol, progesterone, and the antiandro-
gen hydroxyflutamide, were able to activate the mu-
tant AR. The change in ligand specificity of the gene
could possibly affect tumor progression by making the
mutant AR more susceptible to other growth-
stimulatory factors. Androgen receptor mutations are
only found in a minority of (relapsed) prostate cancer
patients, and it is therefore questionable whether the
response pattern reported for CWR22 also applies to
the prostate cancer patient population in general.
In a recent review of model systems of prostate
cancer, three other CWR xenografts (CWR21, CWR31,
and CWR91) were listed, of which no reports are yet
available [29].
The LAPC Series of Xenografts
In 1997, investigators from the Department of
Medicine, UCLA School of Medicine, first described
their achievements in establishing human prostate
cancer xenografts, using SCID mice injected with tu-
mor cell suspensions mixed with Matrigel [30]. They
were able to establish two serially transplantable mod-
els (LAPC-3 and LAPC-4) out of eight attempts with
tumors from different patients with advanced disease.
The LAPC-3 tumor is independent of androgen for its
growth, whereas LAPC-4 shows androgen-responsive
growth: growth is inhibited after castration, followed
by a relapse of growth. However, androgen-
independent LAPC-4 tumors (LAPC-4 AI) do develop
in castrated male mice, although with a prolonged lag
phase. Both xenografts, LAPC-3 and LAPC-4, have a
wild-type hAR and secrete PSA. Also, micrometasta-
ses were detected. More recently, a new xenograft
model, LAPC-9, was presented by the same group
[31]. This tumor is also androgen-responsive and ex-
presses the wild-type hAR. Tumors cease to grow fol-
lowing castration of tumor-bearing mice and remain
dormant for at least 6 months, after which finally
spontaneous, androgen-independent tumors reap-
pear. Studies, similar to the ones performed by us in
the past using the PC-82 tumor [21,32], showed the
survival capacity and androgen responsiveness of
long-term androgen-ablated tumor cells. However, in
contrast to the PC-82 tumor, LAPC-9 tumors showed
androgen-independent growth when tumor cell sus-
267
pensions were injected in female mice, although less
frequently than LAPC-4 [31].
The LuCaP Series of Xenografts
In 1996, Ellis et al. [33] reported on a new androgen-
sensitive prostate tumor xenograft model, designated
LuCaP 23. The tumor originated from lymph nodes
(LuCaP 23.1 and LuCaP 23.8) and liver (LuCaP 23.12)
metastases of a patient with late-stage, hormone-
refractory disease. Tumors were passaged by trans-
planting small fragments of tumor subcutaneously in
BALB/c athymic nude mice. All three xenograft mod-
els secreted PSA. None of the tumors was strictly an-
drogen-dependent, since tumors could be established
in 20–50% of castrated male mice. Interestingly, no
tumors developed in female mice. All LuCaP 23 sub-
lines showed a more or less similar response pattern to
castration, represented by three categories of respond-
ers: minimal responders (showing continued growth),
intermediate responders (showing a small decline in
growth followed immediately by a spontaneous
growth relapse), and prolonged responders (showing
long-term cessation of growth without spontaneous
growth relapse) [33]. This response pattern in many
ways reflects the castration-induced response ob-
served in the PC-346 xenograft.
The MDA Series of Xenografts
Very recently, a set of newly established xenograft
models was introduced in a review article on model
systems of prostate cancer [29]. These xenografts were
developed at the M.D. Anderson Cancer Center
(Houston, TX), and originated from liver metastases
(MDA PCa 31 and 40), an adrenal metastasis (MDA
PCa 43), and a skin metastasis (MDA PCa 44). MDA
PCa 31 and MDA PCa 43 were reported to express AR
and secrete PSA. No further information on the char-
acteristics of these xenografts is yet available.
DISCUSSION
In Vitro Cell Lines From Xenograft Models
In vivo models are essential for the study of the
biological behavior of tumor tissue in its natural (or-
gan) environment that cannot easily be mimicked in
an in vitro setting. Important physiological processes
that are lacking in vitro include three-dimensional
structure, angiogenesis (presence or lack of circulating
activating/inhibiting factors), stromal interaction
(stroma-derived activating/inhibiting factors) influ-
encing tumor development and tumor growth, and
finally, metastatic spread to other tissues. Neverthe-
268 van Weerden et al.
less, the derived in vitro cell lines greatly enhance the
utility of xenografts, as they open many possibilities
for research in a well-controlled system.
For many years, a number of research groups tried
to develop in vitro cell lines for human prostate can-
cer, either by direct culture of patient material or by
cultures of xenograft material. The number of cell lines
established directly from patient material is limited,
and most of them represent the androgen-
independent phenotype of prostate cancer. The LN-
CaP cell line was, until recently, the only androgen-
sensitive human prostate cancer cell line available
[34]. LNCaP cells are known to have a mutated andro-
gen receptor (T877A) which renders these cells re-
sponsive to all kinds of steroids, including antiandro-
gens, which limits their general applicability [35].
Although its tumorigenicity proved rather poor in
nude mice, the LNCaP cell line has been used to create
LNCaP sublines which can be grown in vivo after sub-
cutaneous or orthotopic inoculation [36–38].
Although numerous efforts have been made to gen-
erate in vitro cell lines from various xenograft models,
this has only been succesful in one cell line up to now.
We were able to establish a permanent in vitro cell line
from PC-346, which we designated PC-346C.
Strangely enough, we were not able to establish a per-
manent in vitro cell line from its related xenograft PC-
346B. The PC-346C cell line secretes high levels of PSA
and expresses normal hAR. It hardly grows under an-
drogen-depleted conditions and, like its xenograft
counterpart, mimicks initial androgen-responsiveness
but eventually relapses into androgen independence.
This model system is presently being used to study the
mechanisms of tumor progression both in vivo and in
vitro.
Other efforts from our group have so far resulted in
cultures from PC-295 and PC-310, with a limited pas-
sage capacity of up to five passages. These cultures
were shown to differentiate towards a neuroendocrine
phenotype upon androgen depletion, as was also ob-
served in vivo, and they are, therefore, interesting
models to study the role of neuroendocrine cells in
prostate cancer [23,24].
Recently, two cell lines were developed by the
MDA Group from a single bone metastasis of a hor-
mone-refractory prostate cancer patient [39]. Al-
though from the same patient, the cell lines, MDA PCa
2a and MDA PCa 2b, show different phenotypes with
regard to tumor growth and androgen sensitivity, and
have different genetic characteristics. They both are
androgen-sensitive and secrete PSA. Recently, two
mutations in the ligand-binding domain of the AR
gene of the MDA PCa 2a cell line were described. One
is the LNCaP mutation (T877A), and the other is a
missense mutation (L701H) which is not present in
LNCaP cells. These mutations are shown to reduce the
affinity for androgens such as DHT and R1881, but to
alter the ligand specificity for other nonandrogenic
steroids. Whether these mutations are also present in
their sister cell line MDA PCa 2b has not been re-
ported yet [40].
Metastatic Potential of Xenograft Models
An important factor of morbidity in the prostate
cancer patient population is the preferential metastatic
spread of tumor cells to lymph nodes and bone [41].
Not much is known of the mechanisms involved in
prostate tumor metastasis, mainly because of the lack
of a suitable model system. Subcutaneous, intrave-
nous, or intraperitoneal injections of prostate tumor
cells in the nude mouse have rarely resulted in metas-
tases. In 1992, the technique of orthotopic transplan-
tion was reintroduced as a means of inducing sponta-
neous metastasis originating from the prostate [42,43].
This has resulted in an increased effort to develop
metastatic model systems with a special focus on bone
metastasis. So far, however, results have been very
limited.
All xenograft models established in our laboratory
are being investigated for their metastatic capability
by orthotopic transplantation of tumor fragments in
the dorsolateral lobe of the nude mouse prostate [44].
Orthotopic injection of PC-346C cells into the dorso-
lateral lobe of the mouse prostate results in large tu-
mors within the dorsolateral prostate lobe, without
infiltration of tumor into the seminal vesicles. Animals
die due to obstruction of the urethra within 1 month
after injection of 1 × 106 tumor cells. No gross metas-
tases have been detected yet, although tumor cells
have been identified and recultured from lymph
nodes and lung. These sublines are presently being
tested for their putative increased metastatic potential.
Like many of our colleagues in the field, we are
working hard to develop a spontaneous bone meta-
static model. So far, no spontaneous bone metastasis
has been found. PC-346C cells were able to induce
tumors when injected directly in the mouse tibia bone.
Pettaway et al. [38] reported on the generation of vari-
ants of PC-3 and LNCaP with an increased metastatic
potential, using orthotopic transplantation. Orthotopi-
cally injected metastatic variants of PC-3 cells pro-
duced tumors in lung and bone. Increased incidence
of lymph node metastases was observed with LNCaP
variants. Thalmann et al. [45] were able to select a
LNCaP subline, C4-2, with increased metastatic poten-
tial to lymph nodes and bone, with an incidence of
11–50% upon subcutaneous and orthotopic injection
in nude mice. These model systems are so far the only
models showing prostate tumor metastasis to bone.
Recently, Nemeth et al. [46] reported on a new
 Xenograft Models in Prostate Cancer Research
mouse model for prostate bone metastasis. Human fe-
tal bone was transplanted subcutaneously in nude
mice, and human prostate tumor cells injected intra-
veneously “homed” to the transplanted bone and
formed tumors. Interestingly, they reported that no
tumors developed when murine fetal bone was used,
suggesting that homing of human prostate cancer cells
to bone is human-specific. This observation contra-
dicts the observed metastatic spread of variants of
PC-3 and LNCaP cell lines to bone, as generated by
other investigators [38,46]. However, if these cell lines
are the exceptions to the rule and homing to bone is, in
general, human-specific, then the utility of the (im-
mune-deficient) mouse model may well have reached
its limits as an appropriate model system for the study
of metastatic prostate cancer.
CONCLUSIONS AND FUTURE PERSPECTIVES
In recent years, a considerable number of human
prostate tumor xenografts have been established, as
can be concluded from this review. The different xe-
nograft models represent the various aspects of hu-
man prostate cancer and are excellent tools to study
the various stages of prostate cancer progression.
However, the major drawback of these models is their
lack of spontaneous metastasis to mouse organs such
as bone. Consequently, one should carefully select xe-
nografts that are relevant with regard to the study
objective and select only those that represent the stage
of disease under study. This applies to both funda-
mental as well as preclinical studies.
Besides the significance of each xenograft for the
study of specific issues of human prostate cancer, the
panel of xenografts is important, as it represent the
whole history of clinical behavior of human prostate
cancer from androgen-dependence towards hormone-
refractory disease. The panel offers the possibility to
compare expression of (unknown), genes which will
give insights into the pathways involved in the tran-
sition of human prostate cancer through its various
phenotypic stages [47–50]. This knowledge may ulti-
mately result in new targets and new treatment mo-
dalities.
Finally, similar to the relevance of a broad panel of
xenograft models, it is essential to establish various
metastatic sublines, which preferably follow the pref-
erential spread to lymph nodes and bone as observed
in the patient. Such metastatic model systems will en-
able us to study the requirements for tumor cells to
metastasize and grow in bone, which, hopefully, will
lead to therapies targeting this process.
ACKNOWLEDGMENTS
The authors are indebted to Sigrun Erkens and Cor-
rina M.A. de Ridder for their technical support.
269
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