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The Prostate 31:168–174 (1997)
Orthotopic Implantation of Human Prostate
Cancer Cell Lines: A Clinically Relevant Animal
Model for Metastatic Prostate Cancer
Klaus Rembrink,1* Johannes C. Romijn,1 Theo H. van der Kwast,2
Herbert Rübben,3 and Fritz H. Schröder1
1
Institute of Urology, Erasmus University Rotterdam, Rotterdam, The Netherlands
Institute of Pathology, Erasmus University Rotterdam, Rotterdam, The Netherlands
3
University of Essen Medical School, Essen, Germany
2
BACKGROUND. To study the metastatic behavior of human prostate cancer cell lines, orthotopic injection in nude mice was performed.
METHODS. Local tumor growth, metastasis formation, prostate-specific antigen, and androgen receptor expression were examined.
RESULTS. Hormone-independent cell lines (PC-3, PC-3-125-1L, and TSU-Pr1) were highly
tumorigenic and had a higher rate of lymph node metastasis after orthotopic than after
subcutaneous implantation. PC-3 cell lines also consistently metastasized to the lungs. The
androgen-sensitive LNCaP cell line showed local growth in 7 of 10, and lymph node metastasis in 4 animals. Significant serum PSA levels and strong receptor expression in primary and
metastatic tumor tissues were observed.
CONCLUSIONS. These results demonstrates that orthotopic implantation of human prostate
cancer cell lines, including LNCaP, reproducibly leads to metastasis formation in nude mice.
Prostate 31:168−174, 1997.
© 1997 Wiley-Liss, Inc.
KEY WORDS:
prostate cancer; orthotopic; metastasis; androgen receptor; experimental
animal
INTRODUCTION
Prostatic carcinoma has become the most frequent
cancer in men in Western society during the last decade [1]: 25–40% of all patients already show metastatic disease at the time of diagnosis. At this stage, a
cure cannot be achieved by any treatment modality
and new approaches are needed. A first step towards
more promising therapeutic strategies would be a better understanding of the metastatic process in general.
This can only be studied in detail by the availability of
valid experimental metastatic model systems for prostate cancer.
Metastases of human prostate cancer after subcutaneous (s.c.) implantation in nude mice have only occasionally been reported [2]. Injection into the spleen
of nude mice resulted in a slightly enhanced rate of
metastasis [3,4]. PC-3 cells injected into the tail vein
© 1997 Wiley-Liss, Inc.
while the vena cava inferior was clamped led to tumor
growth in lumbar vertebras, pelvis, and femur [5].
More realistic models of human prostate cancer can
be achieved by orthotopic (o.t.) implantation [6].
Within the last several years this approach has been
used with a variety of other tumors [7]. Recently, two
papers concerning o.t. implantation of human prostate
cancer cell lines were published. Fu et al. [8] implanted
PC-3 and DU145 tumor fragments, obtained after s.c.
tumor growth. Stephenson et al. [9] injected suspensions of PC-3M and LNCaP cells into the nude mouse
prostate. Metastases were found in almost all animals
*Correspondence to: Klaus Rembrink, M.D., University of Essen
Medical School, Institute of Urology, Division of Experimental Oncology, Hufelandstr. 55, 45122 Essen, Germany.
Received 29 November 1995; Accepted 19 March 1996
Orthotopic TX of Prostate Cancer Cells
169
Fig. 1. A: Nude mouse in supine position. Lower abdomen is opened, and the seminal vesicles (SV) are carefully retracted with a sterile
cotton swab. A 30-gauge needle is inserted into the right dorsal lobe of the prostate. B: Whole-mount section of a very large prostatic
tumor (T) grown after orthotopic implantation of 1 × 106 PC-3-125-1L cells. Adjacent to the tumor mass, the empty bladder (B) as well
as the right and left seminal vesicles (SV), partly filled with semen, can be seen.
implanted with PC-3M cells, but in none of the mice
with LNCaP cells. In spite of this negative finding, we
included LNCaP in our studies, because it is the only
hormone-sensitive in vitro cell line presenting the human androgen receptor and secreting prostate-specific
antigen. The aims of this study were: 1) to determine
tumor incidence and development of metastases in
nude mice injected s.c. and o.t. with hormoneindependent and hormone-responsive prostate cancer
cell lines, and 2) to evaluate the expression of marker
proteins like prostate-specific antigen (PSA) and human androgen receptor (hAR) in tumors growing at
different anatomical sites.
MATERIALS AND METHODS
Prostate Cancer Cell Lines
Four human prostate cancer cell lines (PC3, PC3125-1L, TSU-Pr1, and LNCaP-FGC) were used in this
study. The PC-3 cell line, established by Kaighn et al.
[10] from a human bone metastasis, was obtained
from Flow Laboratories (Irvine, Scotland). The
LNCaP-FGC, further referred to as LNCaP, was originally provided by Dr. J. Horoszewicz [11]. The TSUPr1 cell line, established by Iiuzmi et al. [12] from a
lymph node metastasis, was obtained from Dr. Jack
Schalken (University of Nijmegen, Nijmegen, The
Netherlands). The PC3-125-1L cell line was a kind gift
from Dr. J.L. Ware, (Medical College of Virginia/VCU,
Richmond, VA) isolated from a spontaneous lung metastasis of the PC-3 cell line [2].
Cell Culture
TSU-Pr1 and LNCaP cells were maintained in
RPMI-1640 (GIBCO, Burlington, Ontario) + 10% or
7.5% fetal calf serum (FCS), respectively. PC-3 and
PC3-125-1L cells were cultured in Minimal Essential
Medium (GIBCO) + 10% FCS. All media were supplemented with glutamine and penicillin/streptomycin.
Cell cultures were maintained at 37°C in a humidified
incubator in an atmosphere of 5% CO2 in air. At near
170
Rembrink et al.
confluence, cells were washed with PBS, harvested
with trypsin/EDTA, solution and finally suspended in
Ca++- and Mg++-free Hank’s balanced salt solution.
Only single-cell suspensions of >90% viability (Trypan
blue exclusion) were used.
Animals
Athymic male nude mice (NMRI nu/nu, 8–12
weeks old, from the breeding colony at the Erasmus
Experimental Animal Center, Erasmus, The Netherlands) were kept in laminar-flow cage racks under
standardized environmental conditions throughout
the experiments. Sterilized food (RMS-GH, obtained
from Hope Farms, Woerden, The Netherlands) and
acidified drinking water were supplied ad libitum.
Tumor Cell Injection
The mice were anesthesized with tribromoethanol
[13] and placed in supine position. A midline incision
was made in the lower abdomen. Tumor cell suspensions (20 ml) were injected into the dorsal mouse prostate using a 30-gauge needle and a 50-ml glass syringe
(Hamilton Bonaduz AG, Switzerland). A slight elevation of either the right or the left dorsal prostate capsule was considered indicative of correct deposition of
tumor cells (Fig. 1A). The abdomen was closed by
single-stitch sutures using 5-0 silk. Subcutaneous injections (1 × 106 cells) were made unilaterally in the
area of the right shoulder.
Autopsy and Histology
The mice were sacrificed by CO2 at various times
(27–91 days) after tumor cell injection. After careful
inspection of gross tumor growth the prostate, bladder, and seminal vesicles were taken out as one unit.
Iliac and preaortic lymph nodes (in cases of o.t. injection) as well as (ipsi- and contralateral) axillary nodes
(after s.c. injection), lungs, liver, and spine were collected and fixed in 3.7% formalin. After paraffinembedding, specimens were serially sectioned and
stained with hematoxylin and eosin. The human origin of the tumors and metastases was confirmed by
bisbenzimid staining [14].
Prostate-Specific Antigen Serum Analysis
Immediately after sacrifice, 1 ml blood was obtained by heart puncture from each mouse and clotted
in Eppendorf tubes in the refrigerator overnight. The
serum was separated and stored at −20°C. Measurement of human PSA was carried out with an automated microparticle-based enzyme immunoassay
TABLE I. Tumorigenicity and Metastases of Three
Human Hormone-Independent Prostate Cancer Cell
Lines After Subcutaneous Implantation of 1 × 106
Tumor Cells
Cell lines
Days postinjection
Mean tumor weight
Range
Tumorigenicity
Metastases
Lymph nodes
Lungs
Liver or bone
Side effects
PC-3
PC-3-125-1L
TSU-Pr1
45
1,148 mg
470–2,190
5/5
Yes
3/5
1/5
None
None
43
1,386 mg
380–2,130
5/5
Yes
4/5
4/5
None
None
43
1,134 mg
370–1,780
5/5
None
0/5
0/5
None
None
(IMx-MEIA; Abbott Diagnostics, Wiesbaden-Delkenheim, Germany). Sensitivity of this assay was 0.07 ng/
ml. PSA concentration was expressed as nanograms of
PSA per ml of serum.
Immunohistochemistry
Immunostaining with MAb F39.4 (obtained from
the Institute of Pathology, Erasmus University, Rotterdam) for hAR was performed on formalin-fixed 5-mmthick slides, as described previously [15].
RESULTS
Subcutaneous Implantation
With hormone-independent cell lines, subcutaneous implantation always resulted in tumor take in murine hosts. Mean tumor weights were very similar for
the three cell lines (Table I). The tumors were grossly
necrotic, partly surrounded by a solid capsule, and
they demonstrated invasiveness (muscle, lymph
nodes). In about 50% of these tumors, tumorinfiltrated lymph nodes were seen adjacent to the primary tumor, suggesting direct growth infiltration. The
PC-3 cell line showed a metastatic involvement of the
axillary lymph nodes in 3 of 5 mice. The more aggressive subline PC-3-125-1L produced positive nodes in 4
of 5 animals. The lungs were affected in one animal
injected with PC-3 and in 4 animals with PC-3-125-1L
cells. In the mouse with PC-3 lung metastases, only a
few tumor-cell colonies were detected, whereas the
animals inoculated with PC-3-125-1L presented disseminated tumor growth all over their lungs. In contrast, TSU-Pr1 showed no metastatic spread at all after
s.c. injection.
In none of the animals injected with either cell line
were contralateral lymph node metastases detected.
Growth and metastasis of s.c.-injected LNCaP cells
were not tested in parallel, but previous experiments
Orthotopic TX of Prostate Cancer Cells
TABLE II. Dose Dependence of Tumor Take and
Metastasis After Orthotopic Implantation of
TSU-Pr1 Cells
Cell no.
1.25 × 104
2.5 × 104
5 × 104
1.25 × 105
2.5 × 105
5 × 105
1 × 106
2 × 106
Days
Take rate
LN-Metb
40
40
40
27
27
27
27
27
1/3
2/3
1/1a
3/3
3/3
3/3
2/2a
3/3
1/3
2/3
1/1
2/3
0/3
1/3
1/2
2/3
TABLE III. Tumorigenicity and Metastatic Capacity of
Three Human Hormone-Independent Prostate Cancer
Cell Lines After Orthotopic Implantation of 1 × 106
Tumor Cells
Cell lines
a
One and two animals were lost in the first week after injection,
respectively.
b
Lymph node metastasis.
consistently revealed a take rate of about 10%, with no
sign of metastatic spread to the lymph nodes or other
sites.
Orthotopic Implantation
In an attempt to determine the appropriate amount
of tumor cells for o.t. inoculation, we varied the TSUPr1 cell number from 1.25 × 104 to 2 × 106 cells. Tumorigenicity was 100% in all groups except for the
two lowest cell doses. The extent of metastasis was
variable (Table II). No lymph node metastases were
detected after injection of 2.5 × 105 cells, but at a dose
of 2 × 106 cells, metastases were found in 2 of 3 animals. In none of the animals within this experiment
was any sign of metastatic spread to visceral organs
apparent.
Based on these results, we decided to inject a dose
of 1 × 106 tumor cells in further experiments. With all
cell lines used, local tumor growth was observed after
o.t. implantation. Growth rate and growth patterns,
however, appeared to be quite different for the various cell lines. As shown in Table III, intraprostatic
tumor growth was reflected by a considerable weight
increase of the urogenital organs. Local tumors with a
size up to 15 mm in diameter were detected (Fig. 1B).
Invasion into the blood vessels, urethra, bladder, and
seminal vesicles occurred in almost all animals injected with the hormone-independent cell lines (Fig.
2A), causing symptoms like urinary retention, hydronephrosis, and kidney insufficiency, often leading to
early death of the animal. After o.t. injection, positive
lymph nodes were always located in the retroperitoneal space. Metastases were easily detectable macroscopically due to their large size (up to 5 mm) and
induration. On the other hand, small tumor-cell nests
in the subcapsular space of the lymph node were only
171
Days postinjection
Mean weight
P + SV + Ba
Range
Tumorigenicity
Metastases
Lymph nodes
Lungs
Liver or bone
Side effects
PC-3
PC-3-125-1L
TSU-Pr1
43
48
32
782 mg
590–990
5/5
Yes
5/5
5/5
None
Yesb
1,306 mg
630–1,820
5/5
Yes
5/5
5/5
None
Yesc
1,122 mg
830–1,360
5/5
Yes
2/5
0/5
None
Yesd
a
P + SV + B, prostate + seminal vesicles + bladder. Normal
average weight is 350–450 mg.
b
Two mice showed urinary retention; one mouse died on
day 41.
c
Four mice had urinary retention in combination with bilateral
kidney insufficiency.
d
One mouse had urinary retention; two mice died on days 27
and 32.
found after microscopic examination (Fig. 2B). Lung
metastases were rarely diagnosed by macroscopic inspection. Microscopic evaluation mostly showed small
tumor-cell aggregations distributed all through the
lung (Fig. 3A). Lung metastases were always associated with the presence of preaortic lymph node metastases.
Tumor growth after inoculation with PC-3 cells was
always accompanied by local invasion, preaortic
lymph node metastases, and microscopic spread to the
lungs (Fig. 3A). With PC-3-125-1L, thought to be a
more aggressive subline of PC-3, we achieved similar
results (Table III). With TSU-Pr1 cells, the take rate
was 100%, but lymph node metastases were noticed in
only 40% of the animals. Spread of tumor cells to the
lungs never occurred.
With the LNCaP cell line, a take rate of 70% was
obtained 89–91 days after o.t. inoculation. LNCaP tumors expanded by dislodging surrounding structures
rather than invading them. Angioinvasive growth was
occasionally seen. Typical for LNCaP was the presence of grossly hemorrhagic areas within tumors. Tumor growth was observed in 7 mice o.t.-implanted
with LNCaP cells. Four of these showed metastatic
spread to the preaortic nodes.
PSA Measurements and Immunohistochemical
Staining of hAR
PSA was undetectable in the sera of nude mice not
implanted with tumor cells. As expected, we could not
172
Rembrink et al.
Fig. 2. A: Mouse prostate 43 days after intraprostatic injection of 1 × 106 PC-3 cells. Note invasion into seminal vesicles (arrows) (original
magnification × 10). B: Subcapsular metastasis of PC-3 cells in a preaortic lymph node. Below, normal mouse lymphocytes are seen. Human
tumor cells are dislodging the regular lymphatic population (original magnification × 20).
Fig. 3. A: Lung metastasis (arrow) 48 days after orthotopic implantation of PC-3-125-1L cells (original magnification × 20). B: Immunohistochemical staining of human
androgen receptor in a metastatic lymph node of the hormone-sensitive LNCaP-FGC line, 89 days after intraprostatic inoculation. Note large, darkly stained spots representing human tumor-cell nuclei; in contrast, smaller mouse
lymphocytes (top, left) are not stained (original magnification × 40).
detect PSA in the serum of mice inoculated with any
of the hormone-independent tumor cell lines. In contrast, 7 of 10 mice implanted o.t. with LNCaP cells
were found to be PSA-positive. The PSA level ranged
from 3.7–219.0 ng/ml. The highest PSA level was
found in the animal with the largest tumor mass. In
the 3 animals without macroscopic or microscopic LNCaP tumor growth, the PSA concentration was below
detection level (Table IV).
Tumors and metastases of PC-3, PC-3-125-1L, and
TSU-Pr1 did not show detectable hAR expression. In
contrast, the tumors formed by the LNCaP cells were
hAR-positive in all cases, including lymph node metastases (Fig. 3B).
DISCUSSION
The limited availability of metastatic in vivo models
is still a barrier to increasing our knowledge of the
Orthotopic TX of Prostate Cancer Cells
TABLE IV. Tumor Growth, Metastases, and PSA Levels
After Orthotopic Implantation of 1 × 106 Tumor Cells
of the Hormone-Sensitive LNCaP-FGC Line
No.
Days
P + SV + Ba
Growth
LN-Metb
PSA
(ng/ml)
217
218
219
220
221
397
398
399
401
402
89
89
89
89
89
91
91
91
91
91
390 mg
460 mg
910 mg
1,130 mg
570 mg
1,180 mg
980 mg
660 mg
370 mg
470 mg
Yes
No
Yes
Yes
No
Yes
Yes
Yes
No
Yes
Yes
No
No
Yes
No
Yes
Yes
No
No
No
3.7
<0.07
148.0
190.0
<0.07
219.0
164.0
21.0
<0.07
14.7
a
P + SV + B, prostate + seminal vesicles + bladder. Normal
average weight is 350–450 mg.
b
Lymph node metastasis.
process leading to progression and metastasis, especially in the field of prostate cancer. Among other
techniques, subcutaneous, intravenous, or intraperitoneal injections of human cancer cells into nude mice
were the most commonly used in prostate cancer research, but metastases rarely occurred [2]. Implantation into other sites, such as seminal vesicles or spleen,
has resulted in local tumor growth only and in formation of liver metastases, respectively. But these approaches must be considered less physiologically relevant than o.t. implantation [3,16].
The technique of o.t. implantation has already been
shown to be effective in other fields, always resulting
in rapid local tumor growth and often also in metastatic disease [17]. In line with the hypothesis of Paget
[18], stating that tumor growth and metastasis require
the presence of a suitable microenvironment, o.t. implantation might be expected to result in higher tumorigenicity and metastases rates. Recently, two papers concerning orthotopic implantation of human
prostate cancer cell lines were published [8,9]. Our
data confirmed the findings of both groups. Except for
LNCaP, all cell lines produced a tumorigenicity of
100% irrespective of injection site, using 1 × 106 cells.
As shown with TSU-Pr1, as few as 1.25 × 104 cells are
able to develop tumors rapidly after o.t. injection. In
general, metastatic behavior was increased after o.t.
implantation as compared to s.c. injection. As an exception, PC-3-125-1L (thought to be more aggressive
than the parental PC-3) revealed nearly no difference
in this respect. Apparently PC-3-125-1L had already
lost the requirement for specific prostatic factors that
allowed the parental PC-3 and other cell lines to produce metastatic disease. An unexpected finding was
the relatively low metastatic ability of TSU-Pr1 cells, in
173
spite of aggressive local tumor growth. Only 32 days
after o.t. implantation the animals had to be sacrificed,
because almost all of them demonstrated serious illness due to local invasiveness and large tumor size.
Still, relatively few metastases were formed compared
to PC-3 and PC-3-125-1L. Clearly, rapid local tumor
growth and metastatic behavior are not necessarily
related. A similar observation was made after o.t. implantation of the in vivo prostate cancer cell line PC133 (data not shown), which produced very large local
tumors (>13 mm) in all animals but never caused
metastatic disease.
The relevance of the prostate environment for
growth and metastasis of prostate cancer cells is illustrated best by the LNCaP cell line. After s.c. injection
no growth occurred at all, but following o.t. inoculation 7 of 10 animals developed tumors. This result is
similar to the findings of Stephenson et al. [9]. However, no lymphatic spread was noted by these authors
after 40 days, whereas we demonstrated positive
lymph nodes 89–91 days after implantation. To our
knowledge, this is the first report of lymph node metastases formed after injection of LNCaP cells into
nude mice. The metastases found were only occupying a small part in the subcapsular space and had not
yet dislodged all the lymphatic tissue. We believe that,
in addition to lymph node metastasis formation, metastatic spread to the lungs and the bone can also occur
with LNCaP cells, given more time to develop.
LNCaP is the only in vitro cell line expressing
important markers such as PSA and hAR. PSA measurements were positive in all mice exhibiting local
LNCaP tumor growth. The amount of PSA seems to be
a function of the local prostate tumor volume and is
not related to the development of lymph node metastases. PSA is probably a very sensitive tumor marker
in the mouse, because in earlier experiments (not
shown here), significant levels of PSA (up to 1.12 ng/
ml) were measured in mice in which tumors were not
(yet) detectable by histological means.
The immunohistochemical examination for hAR revealed a strong expression pattern not only in the primary tumor but also in the metastatic lymph nodes.
There was no apparent loss of hAR during the metastatic process. In contrast to former assumptions, recent examinations with human material have proven
that hAR is not necessarily lost in metastatic disease,
even in patients presenting hormone-resistant disease
[19]. The unequivocal demonstration of hAR in lymph
node metastases confirms the relevance of the o.t.
model system and the importance of the LNCaP cell
line for the imitation of human metastatic prostate
cancer.
174
Rembrink et al.
CONCLUSIONS
In conclusion, o.t. implantation of human prostate
cancer cells into nude mice leads to the reproducible
formation of lymph node and lung metastases, and is
superior to the s.c. technique. Whereas the implantation site seems to be less critical for the development
of metastatic disease from aggressive cell lines such as
PC-3-125-1L, growth and subsequent metastasis of
other prostate cancer cell lines, such as LNCaP, appear
to be strongly stimulated in a prostatic environment.
During the metastatic process there was no loss of
hAR. Our data show that o.t. implantation with human prostate cancer cell lines, especially LNCaP, is a
very promising approach for studying the mechanisms of human prostate cancer progression and metastasis. Further experiments will be aimed at the isolation of a more metastatic LNCaP subline, still expressing important markers such as PSA and hAR.
6.
7.
8.
9.
10.
11.
12.
ACKNOWLEDGMENTS
13.
We gratefully acknowledge the work of Mrs. Coby
Peekstok in excellently processing all histologies, and
of Dr. M.A. Blankenstein in PSA measurements. K.R.
is the recipient of a research grant of the Dr. Mildred
Scheel Stiftung für Krebsforschung (German Cancer
Foundation).
14.
15.
REFERENCES
1. Boring CC, Squires TS, Tong T: Cancer statistics, 1994. CA 44:7,
1994.
2. Ware JL, Paulson DF, Mickey GH, Webbs KS: Spontaneous metastasis of cells of the human prostate carcinoma cell line PC-3 in
athymic nude mice. J Urol 128:1064–1067, 1982.
3. Shevrin D, Grony K, Kukreja S: Patterns of metastasis by the
human prostate cancer cell line PC-3 in athymic nude mice.
Prostate 15:187–194, 1989.
4. Sherwood E, Ford J, Lee C, Kozlowski J: Therapeutic efficacy of
recombinant tumor necrosis factor alpha in an experimental
model of human prostatic carcinoma. J Biol Response Mod 9:
44–52, 1990.
5. Shevrin D, Kukreja S, Ghosh L, Lad T: Development of skeletal
16.
17.
18.
19.
metastases by human prostate cancer in athymic nude mice.
Clin Exp Metastasis 6:401–409, 1988.
Hoffman RM: Orthotopic is orthodox: Why are orthotopictransplant metastatic models different from all other models? J
Cell Biochem 56:1–3, 1994.
Kubota T: Metastatic models of human cancer xenografted in
the nude mouse: The importance of orthotopic transplantation.
J Cell Biochem 56:4–8, 1994.
Fu X, Herrera H, Hoffman RM: Orthotopic growth and metastasis of human prostate carcinoma in nude mice after transplantation of histological intact tissue. Int J Cancer 52:987–990, 1992.
Stephenson RA, Dinney CN, Gohji K, Ordonez NG, Killion JJ,
Fidler IJ: Metastatic model for human prostate cancer using orthotopic implantation in nude mice. JNCI 84:951–957, 1992.
Kaighn ME, Narayan KS, Ohnuki Y, Lechner J, Jones LW: Establishment of a human prostate cancer cell line (PC-3). Invest
Urol 17:16–23, 1979.
Horoszewicz JS, Leong SS, Kawinski E, Karr JP, Rosenthal H,
Ming-Chu T, Mirand EA, Murphy GP: LNCaP model of human
prostatic carcinoma. Cancer Res 43:1809–1818, 1983.
Iizumi T, Yazuki T, Kanoh S, Kondo I, Koiso K: Establishment of
a new prostatic carcinoma cell line (TSU-Pr1). J Urol 137:1304–
1306, 1987.
Papaioannou VE, Fox JG: Efficacy of tribrimoethanol anesthesia
in mice. Lab Anim Sci 43:189–192, 1993.
Rygaard K: A rapid method for identification of murine cells in
human malignant tumours grown in nude mice. In Rygaard J,
Brunner N, Graen N, Spang-Thompson M (eds): ‘‘ImmuneDeficient Animals in Biomedical Research.’’ Basel: Karger, 1987,
pp 268–272.
Janssen PJ, Brinkmann AO, Boersma WJ, van der Kwast TH:
Immunohistochemical detection of the androgen receptor with
monoclonal antibody F39.4 in routinely processed, paraffinembedded human tissues after microwave pre-treatment. J Histochem Cytochem 42:1169–1175, 1994.
Kozlowski JM, Fidler IJ, Campbell D: Metastatic behavior of
human tumor cell lines grown in nude mouse. Cancer Res 44:
3522–3529, 1984.
Fidler IJ: Rationale and methods for the use of nude mice to
study the biology and therapy of human cancer metastasis. Cancer Metastasis Rev 5:29–49, 1986.
Paget S: The distribution of secondary growths in cancer of the
breast. Cancer Metastasis Rev 1:571–573, 1989.
Ruizeveld de Winter JA, Janssen PJA, Sleddens HMEB, VerleunMooijman MCT, Trapmann J, Brinkmann AO, Santerse AB, Schröder FH, van der Kwast TH: Androgen receptor status in localized and locally progressive hormone refractory human prostate cancer. Am J Pathol 144:735–746, 1994.
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