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Laboratories for Reproductive Biology Departments of Pediatrics (J.-A.T., K.G.H., C.W.G., D.-Y.Z., M.S., E.M.W., F.S.F.), Biochemistry and Biophysics (E.M.W.), Surgery (Y.S., J.L.M.), and Pathology (J.L.M.) and The Lineberger Comprehensive Cancer Center (E.M.W., F.S.F., J.L.M.) School of Medicine The University of North Carolina Chapel Hill, North Carolina 27599 Department of Urology and Medicine (P.G., R.dW.) University of California Davis Medical Center Sacramento, California 95817 Department of Medicine (S.E.H.) The University of Texas Health Science Center San Antonio, Texas 78284-7756 Department of Pathology (T.G.P.) School of Medicine Case Western Reserve University Cleveland, Ohio 44160
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ABSTRACT |
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INTRODUCTION |
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TOPABSTRACTINTRODUCTIONRESULTSDISCUSSIONMATERIALS AND METHODSREFERENCES |
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The first AR missense mutation identified in human prostate cancer was in the LNCaP cell line (1, 2, 3, 4). Subsequently, amino acid substitutions were reported in organ-confined as well as metastatic tumors (5, 6, 7, 8, 9, 10, 11). Of the several AR mutants in prostate cancer that have been characterized thus far, the majority are functional and can mediate androgen-induced transactivation, in contrast to loss of function germ-line AR mutations that cause androgen insensitivity.
CWR22 is an androgen-dependent human prostate cancer xenograft propagated in male nude mice (12, 13). Upon androgen withdrawal, CWR22 prostate-specific antigen mRNA and protein decrease rapidly, cells undergo apoptosis (13, 14), and tumors regress in size; but after several months they recur in the absence of testicular androgens in a manner characteristic of human prostate cancers (13, 14). In this report a mutant AR with altered ligand specificity is identified and shown to be expressed in CWR22 epithelial cells. Ligand-dependent activation of the mutant AR in transient cotransfection assays is compared with that of the mutant AR in LNCaP cells. The CWR22 AR mutant is transcriptionally active in response to testicular androgen like wild type AR but differs from wild type in that it is also activated by the adrenal androgen, dehydroepiandrosterone, the antiandrogen, hydroxyflutamide, and by estradiol and progesterone.
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RESULTS |
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TOPABSTRACTINTRODUCTIONRESULTSDISCUSSIONMATERIALS AND METHODSREFERENCES |
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). A preimmune control panel (Fig. 1C) indicated that
staining in Fig. 1B is AR specific. Hematoxylin-eosin staining (Fig. 1A) illustrates the predominance of epithelial cells with lack of
surrounding stroma. Northern blot analysis of CWR22 total RNA revealed
a 9.6-kb AR mRNA (Fig. 2A) similar in size to the AR
mRNA expressed in benign prostatic hyperplasia tissue (15). In an
RT-PCR assay, expression of AR and prostate-specific antigen (PSA)
mRNAs in CWR22 was similar to that in the androgen-responsive LNCaP
cell line (Fig. 2B) and contrasted with the undetectable or very low
levels in the androgen- unresponsive cell lines, DU145, PC3, and
TSU-PR1. These results are consistent with the androgen-responsive and
relatively differentiated state of CWR22. In mice bearing CWR22
xenografts, Wainstein et al. (13) observed that serum PSA
concentrations correlate with tumor size, and blood levels of PSA
decrease after androgen withdrawal. More recently, Gregory et
al. (14) demonstrated a decrease in CWR22 PSA mRNA levels within
48 h after androgen withdrawal.
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),
suggesting a mismatch in the heteroduplex of wild type and CWR22 DNA.
No similar indication of an AR gene mutation was detected in exons A
through G. Automated sequencing of exon H DNA detected a C to T
mutation at position 4046, numbered according to Lubahn et
al. (16), that changed codon 874 from CAT coding for histidine to
TAT coding for tyrosine (Fig. 3, A and C). The mutation was detected
also by RT-PCR from CWR22 total RNA and manual sequencing of the cDNA
region encoded by exon H. Substitution of T for C 4048 altered a
SfaNI recognition sequence (GCATC to GTATC).
SfaNI did not cut exon H DNA from the original CWR22
xenograft, indicating that the AR mutation was likely present in the
tumor before transplantation. In contrast, SfaNI cut AR exon
H DNA amplified from paraffin-embedded normal testicular tissue of the
same patient, indicating absence of the C to T mutation (data not
shown). Thus, H874Y was a somatic mutation in the prostate tumor not
present in germline DNA.
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-ZAPII
cloning vector (2, 3). AR mutations were constructed in the expression
vector, pCMVhAR (18, 19), and cotransfected into CV1 cells together
with a mouse mammary tumor virus long terminal repeat-luciferase
reporter vector.
CWR22 AR mutant H874Y induced higher luciferase activity than wild type
AR (P < 0.01) at 0.01 nM testosterone (T),
a concentration close to the physiological range of free T in human
blood (20) (Fig. 4). The response of H874Y to
dihydrotestosterone (DHT) at 0.01 nM was similar to that of
wild type AR and about 2-fold more active (P < 0.001)
than LNCaP mutant T877A. Activity induced by 1 nM
androstenedione was similar with H874Y or wild type AR and slightly
higher (P = 0.001) than T877A (Fig. 4). Little
transactivation was observed at lower concentrations of
androstenedione. Because androstenedione has low affinity for the wild
type AR (6), its induction of transcriptional activity with both mutant
and wild type ARs likely resulted from conversion to T and DHT in CV1
cells. Relative responses of wild type and mutant ARs to T and DHT and
to androstenedione were similar.
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). At 100 nM
DHEA, transcriptional activity with H874Y was similar to that of T877A.
DHEA had little agonist activity with wild type AR up to 100
nM, indicating there was minimal conversion of DHEA to T or
DHT in CV1 cells. This concentration of DHEA is similar to that
reported in human prostate tissue as discussed below.
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). Induction of transcription with T877A was also
greater (P < 0.001) than with wild type AR at 1
nM estradiol but was only about half that of H874Y. The
physiological level of total serum estradiol in the adult male is
approximately 0.1 nM whereas free estradiol is 1–2% of
the total (20). There was little increase in transcription at 0.01
nM estradiol suggesting H874Y would not be activated in the
adult male; however, aromatase activity in human prostate may cause
tissue levels to be higher than in serum. The CWR22 mutant AR
transcriptional response to 1 nM progesterone was greater
(P < 0.01) than that of the wild type AR but less
(P < 0.01) than the LNCaP mutant (Fig. 6). Total serum
progesterone concentration in adult males, 
1 nM, is
similar to that in females during the follicular phase of the menstrual
cycle (20) and may be sufficient to activate a mutant AR in
prostate.
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). However, at 10 nM hydroxyflutamide,
H874Y and T877A transcriptional activities were 4- and 6-fold greater
(P < 0.001) than that of wild type AR (Fig. 6
).
Maximal luciferase activities stimulated by DHEA and hydroxyflutamide
with the mutant ARs were 15–20% of activities induced by T and
DHT.
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DISCUSSION |
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TOPABSTRACTINTRODUCTIONRESULTSDISCUSSIONMATERIALS AND METHODSREFERENCES |
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DHEA-induced transactivation by H874Y and T877A was highest with 100
nM DHEA, a concentration likely maintained in human
prostate tissue by adrenal secretion of DHEA and DHEA-sulfate. Total
serum DHEA in adult males (mean, 25 nM) is within the
range of total serum testosterone. However, relatively more serum DHEA
should be available to prostate cells because it circulates loosely
bound to albumin (20). Serum DHEA-sulfate can be 300–500 times the
concentration of DHEA, and the sulfatase present in human prostate
converts DHEA-sulfate to DHEA (21). Mean concentrations of DHEA-sulfate
and DHEA in nonhyperplastic tissue specimens obtained by total
prostatectomy were 300 and 90 pmol/mg DNA, respectively (equivalent
to 300 and 90 nM, assuming 1 mg DNA/g tissue) (21). In a
patient undergoing androgen withdrawal therapy by castration or LH
suppression, DHEA could be in a concentration range sufficient to
activate mutant ARs such as H874Y, T877A, or the AR mutant V715M
identified in a prostate cancer bone marrow metastasis by Culig
et al. (6). In human prostate, low levels of
4,5 isomerase activity limit DHEA conversion to
4 androgens (21). However, small amounts of DHT were
formed from DHEA and DHEA-sulfate in human benign prostatic hyperplasia
tissue (22).
Estradiol derived from peripheral conversion of adrenal androstenedione is a potential agonist for mutant ARs in prostate cancer. Estradiol induced an increase in luciferase activity with both the CWR22 and LNCaP mutants at a concentration of 10-9 M, less than the amount required for half-maximal stimulation of LNCaP cell growth (10-8 M). In our transcription assays, estradiol was a greater agonist with the CWR22 mutant AR than with the LNCaP mutant. However, CWR22 does not grow in female nude mice, indicating growth stimulation by H874Y is not activated by female mouse levels of estradiol. It was assumed the LNCaP mutant AR mediates the growth-stimulating effect of estradiol on LNCaP cells (2, 3, 4). However, a recent report demonstrated that LNCaP cells also contain estrogen receptors based on ligand binding and immunocytochemical assays (23). The antiestrogen ICI-182,780 inhibited estradiol stimulation of growth but did not inhibit growth induced by DHT, suggesting the growth effect of estradiol was mediated by estrogen receptors rather than by the T877A mutant AR. It remains to be determined whether higher concentrations of estradiol stimulate CWR22 tumor growth by way of the mutant AR. Estrogen receptors were not detected by immunostaining CWR22 tumor cells with three different antibodies to the human estrogen receptor, a finding in agreement with other immunohistochemical studies showing weak or undetectable staining of estrogen receptors in benign prostatic hyperplasia or prostate cancer epithelial cells (24). It is not yet established whether the human homolog of rat prostate estrogen receptor ß (25) is expressed in CWR22.
Hydroxyflutamide is the active metabolite of flutamide, an androgen antagonist used in the treatment of prostate cancer. Both H874Y and T877A mediated increased transcriptional activity at 10 nM hydroxyflutamide whereas there was no effect with wild type AR. Another AR mutant in human prostate cancer, V715M, was unresponsive to 100 nM hydroxyflutamide (6) but responded 2-fold greater than wild type AR to micromolar concentrations as did the prostate cancer AR mutant V730M (26). In the absence of androgen, hydroxyflutamide is an agonist with wild type AR at concentrations of 1 µM or higher (27, 28). As noted by Wong et al. (27) and by Kemppainen and Wilson (29), this could be of clinical importance in the regression of prostate cancer observed in some patients whose flutamide treatment has been withdrawn (30, 31). Prostate cancer patients treated with high-dose flutamide can have plasma levels of hydroxyflutamide greater than 1 µM. During androgen withdrawal therapy, hydroxyflutamide could function as an agonist when concentrations greatly exceed the concentration of androgen, although its affinity for wild type AR is at least 1 order of magnitude less than that of DHT (6, 27). AR mutants such as H874Y and T877A with enhanced responsiveness to hydroxyflutamide would be more active than wild type AR in mediating growth effects of hydroxyflutamide on prostate cancer cells. Half-maximal stimulation of LNCaP cell growth in culture was achieved with 10-8 M hydroxyflutamide (data not shown), in agreement with other studies (32, 33, 34). In general, the relative growth-stimulating activity of an AR agonist correlates with its binding affinity for AR and induction of AR-mediated transactivation in transient cotransfection assays (2, 3, 4, 32, 33, 34, 35, 36, 37).
Of the different AR missense mutations reported in clinical prostate
cancer (reviewed in Refs. 11 and 38 and B. Gottlieb, Androgen Receptor
Gene Mutations Database), most that were characterized functionally
retain androgen-dependent transcriptional activity. V730M AR from a
stage B, organ-confined prostate cancer had no loss of androgen binding
or transcriptional activity (5, 39). The LNCaP cell line containing AR
mutant T877A was derived from a lymph node metastasis. Three additional
mutations, V715M, T877S, and H874Y, were identified in bone metastases
(6, 7). Each of the mutants exhibited essentially normal
transactivation in response to DHT. The V715M mutant showed no steroid
binding abnormality but had increased transcriptional activity relative
to wild type AR in response to the adrenal androgens, DHEA and
androstenedione (6). Mutants H874Y and T877S were reported to have
enhanced transcriptional activity with progesterone and estradiol at
concentrations of 10 and 100 nM (7). Mutant Q919R at the
carboxyl terminus of AR remained functional in response to DHT, with
only partial loss of activity (40). The only loss of function mutations
reported in association with prostate cancer are a single base change
that introduced a stop codon (Trp 794 to stop) in the steroid-binding
domain (9) and C619Y in exon C close to the DNA-binding domain (40). By
analogy with the crystal structures of the thyroid hormone receptor and
retinoic acid receptor 
ligand-binding domains (41, 42), AR ligands
are buried in a hydrophobic pocket formed by several 
-helices folded
in a way such that widely separated amino acids are brought together to
form the pocket. AR amino acids 874 and 877 are located within a
conserved heptad repeat of hydrophobic residues (38, 43) in -helix
11 (41).
Since AR is required for androgen-dependent growth of prostate cancer cells, tumors may select for AR mutants that retain activity. Most prostate cancers that relapse during androgen withdrawal therapy express AR at levels similar to androgen-dependent prostate cancer (Refs. 44 and 45 and J. L. Mohler, unpublished). Functional AR mutants with enhanced responsiveness to adrenal androgens and/or antiandrogens could have an active role in the survival and recurrent growth of prostate cancer cells. Moreover, the altered spectrum of ligand responsiveness may potentiate ligand-independent activation of AR in relapsed prostate cancers with increased kinase activities (46, 47, 48).
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MATERIALS AND METHODS |
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TOPABSTRACTINTRODUCTIONRESULTSDISCUSSIONMATERIALS AND METHODSREFERENCES |
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Human Prostate Cancer Tissue (CWR22)
CWR22 is an androgen-dependent, human prostatic carcinoma
xenograft established from a transurethral resection and grown
subcutaneously in male athymic nude mice, each containing a 12.5-mg
testosterone pellet (Innovative Research, Sarasota, FL) implanted
subcutaneously (12). The tumor was stage D with osseous metastases,
Gleason grade 9. After removal of CWR22 xenografts from nude mice,
tumor tissue was frozen immediately in liquid nitrogen and stored at
-80 C.
Immunocytochemistry
Small pieces of fresh tumor tissue (1 cm2 x 0.3
cm thick) were frozen in isopentane precooled in liquid nitrogen on
brass mounts using minced rat liver cushions as an adhesive. Tissue was
stored in liquid nitrogen until sectioned at 6 µm thickness using a
cryostat at -31 C. Preparation of antibody AR-52 and the method for
immunostaining were as described previously (15, 49, 50).
Nucleic Acid Extraction and Northern Hybridization of Human
AR
Frozen CWR22 tumor tissue was thawed, finely minced with
scalpels, digested with proteinase K at 37 C overnight, and DNA
extracted with phenol-chloroform. RNA was removed by RNase digestion
and phenol-chloroform extraction. For extraction of normal testicular
tissue DNA from the same patient, 6 µm thick sections were cut from
paraffin blocks and deparaffinized in xylene. Tissue was digested with
proteinase K, and DNA was isolated by NaCl deproteinization (10). Total
RNA was extracted by a single-step method using acid-guanidinium
thiocyanate-phenol-chloroform (51). Northern hybridization of AR mRNA
was as described by Quarmby et al. (52).
Cell Lines
The human prostate tumor cell lines LNCaP, DU145, and PC-3 were
obtained from the American Type Culture Collection (Rockville, MD) and
maintained according to their instructions. The TSU-Pr1 cell line was
generously provided by Dr. Edward Gelmann (Georgetown University School
of Medicine, Washington DC) and maintained in DMEM supplemented with
10% FBS. Cultures were harvested for RNA analysis at approximately
80% confluence.
RT-PCR Analysis of Human AR and PSA mRNA
RT-PCR analysis of AR mRNA was done essentially as
previously described (45). Briefly, total RNA was extracted from the
cell lines and xenografts using guanidinium isothiocyanate and CsCl
density gradient centrifugation. Total RNA was transcribed to cDNA
using RT primed with random hexamers. Complementary DNA equivalent to
25 ng of the initial total RNA was subjected to RT-PCR using the
following oligonucleotides as primers for the AR
transcripts: PG45 (upstream, sense) 5'-CCTGATCTGTGGAGATGAAGCTTC-3' and
PG46 (downstream, antisense) 5'-TGTCGTGTCCAGCACACACTACAC-3'. These
primers are located in exons B and D, respectively, and create a 495-bp
PCR product from cDNA. The oligonucleotide PG42
(5'-TGGGAGCCCGGAAGCTGAAGAAAC-3') (spanning the junction of exons C and
D) is internally positioned to the two primers and served as the probe
for detecting the products in Southern blotting. PCR was run for 36
cycles under the conditions of 95 C (30 sec), 60 C (30 sec), and 72 C
(30 sec). The 36 cycles were followed by a 10-min extension phase at 72
C. Southern blotting of the PCR products was done as previously
reported using a nonisotopic enzyme chemiluminescent detection system
(53). For analysis of expression of PSA transcripts, the primers
PSA-494 (upstream, sense) 5'-TACCCACTGCATCAGGAACA-3' and PSA-894
(downstream, antisense) 5'-GTCCAGCGTCCAGCACACAG-3' were used to create
a 421-bp product (54). PCR reaction parameters were 94 C for 15 sec, 60
C for 15 sec, and 72 C for 45 sec for 35 cycles followed by a 10-min
extension at 72 C. The oligonucleotide used as the probe for Southern
blotting of PSA RT-PCR products was PSA-559
5'-ACACAGGCCAGGTATTTCAG-3'. Sample loadings were adjusted to equivalent
levels of c-N-ras as previously described and validated by Fishman
et al. (55)
Analysis of the AR Gene in CWR22
Amplification of AR gene DNA.
Exons B through H were amplified individually using GC-clamped intron
primers. Due to its large size, exon A was amplified in four fragments
using exon primers (16, 17). PCR was performed for 30 cycles of
denaturation (95 C for 1 min), annealing (60 C for 1 min), and
extension (72 C for 2 min). In each PCR a negative control containing
no DNA was included. PCR product size was verified by electrophoresis
on 1% agarose gels in Tris-borate buffer containing 2 mM
EDTA, 50 mM Tris base, 50 mM boric acid, pH 8.0
(TBE), and ethidium bromide staining.
Denaturing Gradient Gel Electrophoresis.
Electrophore-sis of amplified AR was performed on denaturing
gradient gels as described (17). Heteroduplexes for each exon were
formed by mixing equal amounts of test sample and wild type PCR
products. DNA was denatured at 95 C and reannealed by slow cooling to
room temperature.
RT-PCR for Splicing Errors.
Total RNA (1 µg) was reverse transcribed into cDNA using AMV RT and
oligo (dT)20 primer. Primers for PCR were chosen to amplify
all cDNA splice sites from exons A through H, a region of 1296 bp.
Amplified AR cDNA fragments were electrophoresed in 1% agarose gels in
TBE and stained with ethidium bromide.
Restriction Site Analysis.
Approximately 200 ng of PCR product of exon H amplification were
incubated with 2 U of the restriction enzyme SfaNI in a
total volume of 20 µl of NE Buffer 3 for 16 h as recommended by
New England Biolabs (Beverley, MA). After incubation, the enzyme was
heat inactivated and the DNA analyzed by electrophoresis in a 1%
agarose gel in TBE buffer.
Subcloning and Sequencing the CWR22 Mutation
An aliquot of the PCR reaction used in denaturing gradient gel
electrophoresis was chloroform extracted and ethanol precipitated. The
DNA ends were made blunt with Klenow fragment (Promega, Madison WI) and
phosphorylated with T4 kinase (Promega). These fragments were cloned
into pBluescript II SK- (Stratagene, La Jolla, CA) cut with
EcoRV (Promega), and dephosphorylated with calf intestinal
phosphatase (Boehringer Mannheim, Indianapolis, IN).
DNA sequence was obtained from four individual clones using double-stranded sequencing with Taq Dideoxy Terminator and an Applied Biosystems 373A automated DNA sequencer (Foster City, CA). Standard M13 vector forward and reverse primers were used. Sequence for both strands of each clone was analyzed using a Genetics Computer Group (GCG) program (Madison, WI).
Construction of Mutant AR Expression Vectors
The CWR22 mutant AR H874Y vector was constructed in pCMVhAR
using the two-step PCR method as described (17, 18). LNCaP AR was
cloned from an LNCaP cDNA library in Zap-II (2, 3). The LNCaP
mutant AR T877A cDNA (2) HindIII/BamHI fragment
replaced the wild type HindIII/BamHI fragment in
pCMVhAR. Expression vector constructs were analyzed by DNA
sequencing.
Transcription Assay
Transcriptional activities of wild type and mutant ARs were
analyzed by transient cotransfection of African green monkey CV1 cells
using a mouse mammary tumor virus long terminal repeat-luciferase
reporter vector as described (27). CV1 cells were maintained at 37 C
under 5% CO2 in high glucose DMEM with 4.5 g/liter glucose
supplemented with 5% FBS. On the day before transfection, 4.5 x
105 cells per 6-cm culture dish were grown in the same
medium for about 20 h until 70–80% confluent. Expression vector
DNA (100 ng) and reporter vector DNA (2.5 µg) were transfected into
CV1 cells using CaPO4 DNA precipitation (56). Cells were
incubated 40 h in serum-free medium with and without steroids with
a change in medium at 24 h and harvested on the plate using lysis
buffer (Ligand Pharmaceuticals Inc., San Diego, CA). The luciferase
assay was performed as described (27). Background activity was defined
as light units determined in the presence of the AR expression vector
and the absence of ligand. Stimulation of luciferase activity is
expressed as fold increase over background based on five or more
independent experiments. Background activities of the AR expression
vectors in light units were: wild type hAR 1817 ± 521, H874Y
2904 ± 1039, and T877A 3718 ± 2153. Student’s t
test and the computer program Sigmastat were used for statistical
analysis. Data are expressed as the means ± SEM of
three to 11 experiments.
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ACKNOWLEDGMENTS |
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FOOTNOTES |
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This work was supported by NIH Grants AG-11343, HD-04466, P30-HD-18968 (DNA, Cell Culture and Histochemistry Cores), T32-HD-07315 (J-A.T. and C.W.G.), CA-57179 (T.G.P.) and CA-55792, CA-55792 (R.W. de V.W.).
1 Cooperative Network for Molecular and Genetic Markers for Prostate
Cancer, National Cancer Institute. 
Received for publication September 3, 1996. Revision received December 16, 1996. Accepted for publication January 15, 1997.
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TOPABSTRACTINTRODUCTIONRESULTSDISCUSSIONMATERIALS AND METHODSREFERENCES |
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B. He, R. T. Gampe Jr., A. T. Hnat, J. L. Faggart, J. T. Minges, F. S. French, and E. M. Wilson Probing the Functional Link between Androgen Receptor Coactivator and Ligand-binding Sites in Prostate Cancer and Androgen Insensitivity J. Biol. Chem., March 10, 2006; 281(10): 6648 - 6663. [Abstract] [Full Text] [PDF]
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J. Duff and I. J. McEwan Mutation of Histidine 874 in the Androgen Receptor Ligand-Binding Domain Leads to Promiscuous Ligand Activation and Altered p160 Coactivator Interactions Mol. Endocrinol., December 1, 2005; 19(12): 2943 - 2954. [Abstract] [Full Text] [PDF]
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C. E. Bohl, D. D. Miller, J. Chen, C. E. Bell, and J. T. Dalton Structural Basis for Accommodation of Nonsteroidal Ligands in the Androgen Receptor J. Biol. Chem., November 11, 2005; 280(45): 37747 - 37754. [Abstract] [Full Text] [PDF]
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J. T. Arnold and M. R. Blackman Does DHEA Exert Direct Effects on Androgen and Estrogen Receptors, and Does It Promote or Prevent Prostate Cancer? Endocrinology, November 1, 2005; 146(11): 4565 - 4567. [Full Text] [PDF]
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F. Chen, K. Knecht, E. Birzin, J. Fisher, H. Wilkinson, M. Mojena, C. T. Moreno, A. Schmidt, S.-i. Harada, L. P. Freedman, et al. Direct Agonist/Antagonist Functions of Dehydroepiandrosterone Endocrinology, November 1, 2005; 146(11): 4568 - 4576. [Abstract] [Full Text] [PDF]
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M. A. Titus, M. J. Schell, F. B. Lih, K. B. Tomer, and J. L. Mohler Testosterone and Dihydrotestosterone Tissue Levels in Recurrent Prostate Cancer Clin. Cancer Res., July 1, 2005; 11(13): 4653 - 4657. [Abstract] [Full Text] [PDF]
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M. A. Titus, C. W. Gregory, O. H. Ford III, M. J. Schell, S. J. Maygarden, and J. L. Mohler Steroid 5{alpha}-Reductase Isozymes I and II in Recurrent Prostate Cancer Clin. Cancer Res., June 15, 2005; 11(12): 4365 - 4371. [Abstract] [Full Text] [PDF]
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 Z Culig, H Steiner, G Bartsch, and A Hobisch Mechanisms of endocrine therapy-responsive and -unresponsive prostate tumours Endocr. Relat. Cancer, June 1, 2005; 12(2): 229 - 244. [Abstract] [Full Text] [PDF]
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J. Huang, J. L. Yao, L. Zhang, P. A. Bourne, A. M. Quinn, P. A. di Sant'Agnese, and J. E. Reeder Differential Expression of Interleukin-8 and Its Receptors in the Neuroendocrine and Non-Neuroendocrine Compartments of Prostate Cancer Am. J. Pathol., June 1, 2005; 166(6): 1807 - 1815. [Abstract] [Full Text] [PDF]
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 L. Gaughan, I. R. Logan, D. E. Neal, and C. N. Robson Regulation of androgen receptor and histone deacetylase 1 by Mdm2-mediated ubiquitylation Nucleic Acids Res., January 7, 2005; 33(1): 13 - 26. [Abstract] [Full Text] [PDF]
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Y. B. Wetherill, N. L. Fisher, A. Staubach, M. Danielsen, R. W. de Vere White, and K. E. Knudsen Xenoestrogen Action in Prostate Cancer: Pleiotropic Effects Dependent on Androgen Receptor Status Cancer Res., January 1, 2005; 65(1): 54 - 65. [Abstract] [Full Text] [PDF]
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H. I Scher, G. Buchanan, W. Gerald, L. M Butler, and W. D Tilley Targeting the androgen receptor: improving outcomes for castration-resistant prostate cancer Endocr. Relat. Cancer, September 1, 2004; 11(3): 459 - 476. [Abstract] [Full Text] [PDF]
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R. J. Jin, Y. Wang, N. Masumori, K. Ishii, T. Tsukamoto, S. B. Shappell, S. W. Hayward, S. Kasper, and R. J. Matusik NE-10 Neuroendocrine Cancer Promotes the LNCaP Xenograft Growth in Castrated Mice Cancer Res., August 1, 2004; 64(15): 5489 - 5495. [Abstract] [Full Text] [PDF]
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 C. A. Heinlein and C. Chang Androgen Receptor in Prostate Cancer Endocr. Rev., April 1, 2004; 25(2): 276 - 308. [Abstract] [Full Text] [PDF]
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C. W. Gregory, X. Fei, L. A. Ponguta, B. He, H. M. Bill, F. S. French, and E. M. Wilson Epidermal Growth Factor Increases Coactivation of the Androgen Receptor in Recurrent Prostate Cancer J. Biol. Chem., February 20, 2004; 279(8): 7119 - 7130. [Abstract] [Full Text] [PDF]
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J. L. Mohler, C. W. Gregory, O. H. Ford III, D. Kim, C. M. Weaver, P. Petrusz, E. M. Wilson, and F. S. French The Androgen Axis in Recurrent Prostate Cancer Clin. Cancer Res., January 15, 2004; 10(2): 440 - 448. [Abstract] [Full Text] [PDF]
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L.-N. Song, M. Coghlan, and E. P. Gelmann Antiandrogen Effects of Mifepristone on Coactivator and Corepressor Interactions with the Androgen Receptor Mol. Endocrinol., January 1, 2004; 18(1): 70 - 85. [Abstract] [Full Text] [PDF]
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 M. Fu, M. Rao, C. Wang, T. Sakamaki, J. Wang, D. Di Vizio, X. Zhang, C. Albanese, S. Balk, C. Chang, et al. Acetylation of Androgen Receptor Enhances Coactivator Binding and Promotes Prostate Cancer Cell Growth Mol. Cell. Biol., December 1, 2003; 23(23): 8563 - 8575. [Abstract] [Full Text] [PDF]
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T. Hara, K. Nakamura, H. Araki, M. Kusaka, and M. Yamaoka Enhanced Androgen Receptor Signaling Correlates with the Androgen-refractory Growth in a Newly Established MDA PCa 2b-hr Human Prostate Cancer Cell Subline Cancer Res., September 1, 2003; 63(17): 5622 - 5628. [Abstract] [Full Text] [PDF]
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 W. G. Nelson, A. M. De Marzo, and W. B. Isaacs Prostate Cancer N. Engl. J. Med., July 24, 2003; 349(4): 366 - 381. [Full Text] [PDF]
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 D. J. Lamb, E. Puxeddu, N. Malik, D. L. Stenoien, R. Nigam, G. Y. Saleh, M. Mancini, N. L. Weigel, and M. Marcelli Molecular Analysis of the Androgen Receptor in Ten Prostate Cancer Specimens Obtained Before and After Androgen Ablation J Androl, March 1, 2003; 24(2): 215 - 225. [Abstract] [Full Text] [PDF]
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T. H. Thin, L. Wang, E. Kim, L. L. Collins, R. Basavappa, and C. Chang Isolation and Characterization of Androgen Receptor Mutant, AR(M749L), with Hypersensitivity to 17-beta Estradiol Treatment J. Biol. Chem., February 21, 2003; 278(9): 7699 - 7708. [Abstract] [Full Text] [PDF]
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B. Comuzzi, L. Lambrinidis, H. Rogatsch, S. Godoy-Tundidor, N. Knezevic, I. Krhen, Z. Marekovic, G. Bartsch, H. Klocker, A. Hobisch, et al. The Transcriptional Co-Activator cAMP Response Element-Binding Protein-Binding Protein Is Expressed in Prostate Cancer and Enhances Androgen- and Anti-Androgen-Induced Androgen Receptor Function Am. J. Pathol., January 1, 2003; 162(1): 233 - 241. [Abstract] [Full Text] [PDF]
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R. M. Adam, J. Kim, J. Lin, A. Orsola, L. Zhuang, D. C. Rice, and M. R. Freeman* Heparin-Binding Epidermal Growth Factor-Like Growth Factor Stimulates Androgen-Independent Prostate Tumor Growth and Antagonizes Androgen Receptor Function Endocrinology, December 1, 2002; 143(12): 4599 - 4608. [Abstract] [Full Text] [PDF]
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C. G. Tepper, D. L. Boucher, P. E. Ryan, A.-H. Ma, L. Xia, L.-F. Lee, T. G. Pretlow, and H.-J. Kung Characterization of a Novel Androgen Receptor Mutation in a Relapsed CWR22 Prostate Cancer Xenograft and Cell Line Cancer Res., November 15, 2002; 62(22): 6606 - 6614. [Abstract] [Full Text] [PDF]
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 M. Maggiolini, A. Vivacqua, A. Carpino, D. Bonofiglio, G. Fasanella, M. Salerno, D. Picard, and S. Ando The Mutant Androgen Receptor T877A Mediates the Proliferative but Not the Cytotoxic Dose-Dependent Effects of Genistein and Quercetin on Human LNCaP Prostate Cancer Cells Mol. Pharmacol., November 1, 2002; 62(5): 1027 - 1035. [Abstract] [Full Text] [PDF]
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 E. P. Gelmann Molecular Biology of the Androgen Receptor J. Clin. Oncol., July 1, 2002; 20(13): 3001 - 3015. [Abstract] [Full Text] [PDF]
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 J. Raber, G. Bongers, A. LeFevour, M. Buttini, and L. Mucke Androgens Protect against Apolipoprotein E4-Induced Cognitive Deficits J. Neurosci., June 15, 2002; 22(12): 5204 - 5209. [Abstract] [Full Text] [PDF]
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C.-Y. Chang and D. P. McDonnell Evaluation of Ligand-Dependent Changes in AR Structure Using Peptide Probes Mol. Endocrinol., April 1, 2002; 16(4): 647 - 660. [Abstract] [Full Text] [PDF]
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B. He, J. T. Minges, L. W. Lee, and E. M. Wilson The FXXLF Motif Mediates Androgen Receptor-specific Interactions with Coregulators J. Biol. Chem., March 15, 2002; 277(12): 10226 - 10235. [Abstract] [Full Text] [PDF]
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X.-B. Shi, A.-H. Ma, L. Xia, H.-J. Kung, and R. W. de Vere White Functional Analysis of 44 Mutant Androgen Receptors from Human Prostate Cancer Cancer Res., March 1, 2002; 62(5): 1496 - 1502. [Abstract] [Full Text] [PDF]
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D. Kim, C. W. Gregory, F. S. French, G. J. Smith, and J. L. Mohler Androgen Receptor Expression and Cellular Proliferation During Transition from Androgen-Dependent to Recurrent Growth after Castration in the CWR22 Prostate Cancer Xenograft Am. J. Pathol., January 1, 2002; 160(1): 219 - 226. [Abstract] [Full Text] [PDF]
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 M. E. Grossmann, H. Huang, and D. J. Tindall Androgen Receptor Signaling in Androgen-Refractory Prostate Cancer J Natl Cancer Inst, November 21, 2001; 93(22): 1687 - 1697. [Abstract] [Full Text] [PDF]
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C. W. Gregory, B. He, R. T. Johnson, O. H. Ford, J. L. Mohler, F. S. French, and E. M. Wilson A Mechanism for Androgen Receptor-mediated Prostate Cancer Recurrence after Androgen Deprivation Therapy Cancer Res., June 1, 2001; 61(11): 4315 - 4319. [Abstract] [Full Text] [PDF]
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G. Buchanan, N. M. Greenberg, H. I. Scher, J. M. Harris, V. R. Marshall, and W. D. Tilley Collocation of Androgen Receptor Gene Mutations in Prostate Cancer Clin. Cancer Res., May 1, 2001; 7(5): 1273 - 1281. [Abstract] [Full Text]
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C. W. Gregory, R. T. Johnson Jr., J. L. Mohler, F. S. French, and E. M. Wilson Androgen Receptor Stabilization in Recurrent Prostate Cancer Is Associated with Hypersensitivity to Low Androgen Cancer Res., April 1, 2001; 61(7): 2892 - 2898. [Abstract] [Full Text]
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G. Buchanan, M. Yang, J. M. Harris, H. S. Nahm, G. Han, N. Moore, J. M. Bentel, R. J. Matusik, D. J. Horsfall, V. R. Marshall, et al. Mutations at the Boundary of the Hinge and Ligand Binding Domain of the Androgen Receptor Confer Increased Transactivation Function Mol. Endocrinol., January 1, 2001; 15(1): 46 - 56. [Abstract] [Full Text]
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J.-a. Tan, S. H. Hall, P. Petrusz, and F. S. French Thyroid Receptor Activator Molecule, TRAM-1, Is an Androgen Receptor Coactivator Endocrinology, September 1, 2000; 141(9): 3440 - 3450. [Abstract] [Full Text] [PDF]
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C. I. Truica, S. Byers, and E. P. Gelmann {beta}-Catenin Affects Androgen Receptor Transcriptional Activity and Ligand Specificity Cancer Res., September 1, 2000; 60(17): 4709 - 4713. [Abstract] [Full Text]
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 J. L. Holleran, C. J. Miller, and L. A. Culp Tracking Micrometastasis to Multiple Organs with lacZ-tagged CWR22R Prostate Carcinoma Cells J. Histochem. Cytochem., May 1, 2000; 48(5): 643 - 652. [Abstract] [Full Text]
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S. McDonald, L. Brive, D. B. Agus, H. I. Scher, and K. R. Ely Ligand Responsiveness in Human Prostate Cancer: Structural Analysis of Mutant Androgen Receptors from LNCaP and CWR22 Tumors Cancer Res., May 1, 2000; 60(9): 2317 - 2322. [Abstract] [Full Text]
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J.-a. Tan, S. H. Hall, K. G. Hamil, G. Grossman, P. Petrusz, J. Liao, K. Shuai, and F. S. French Protein Inhibitor of Activated STAT-1 (Signal Transducer and Activator of Transcription-1) Is a Nuclear Receptor Coregulator Expressed in Human Testis Mol. Endocrinol., January 1, 2000; 14(1): 14 - 26. [Abstract] [Full Text]
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B. He, J. A. Kemppainen, J. J. Voegel, H. Gronemeyer, and E. M. Wilson Activation Function 2 in the Human Androgen Receptor Ligand Binding Domain Mediates Interdomain Communication with the NH2-terminal Domain J. Biol. Chem., December 24, 1999; 274(52): 37219 - 37225. [Abstract] [Full Text] [PDF]
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L. V. Nazareth, D. L. Stenoien, W. E. Bingman III, A. J. James, C. Wu, Y. Zhang, D. P. Edwards, M. Mancini, M. Marcelli, D. J. Lamb, et al. A C619Y Mutation in the Human Androgen Receptor Causes Inactivation and Mislocalization of the Receptor with Concomitant Sequestration of SRC-1 (Steroid Receptor Coactivator 1) Mol. Endocrinol., December 1, 1999; 13(12): 2065 - 2075. [Abstract] [Full Text]
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D. B. Agus, C. Cordon-Cardo, W. Fox, M. Drobnjak, A. Koff, D. W. Golde, and H. I. Scher Prostate Cancer Cell Cycle Regulators: Response to Androgen Withdrawal and Development of Androgen Independence J Natl Cancer Inst, November 3, 1999; 91(21): 1869 - 1876. [Abstract] [Full Text] [PDF]
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B. B. Yeap, R. G. Krueger, and P. J. Leedman Differential Posttranscriptional Regulation of Androgen Receptor Gene Expression by Androgen in Prostate and Breast Cancer Cells Endocrinology, July 1, 1999; 140(7): 3282 - 3291. [Abstract] [Full Text]
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M.-E. Taplin, G. J. Bubley, Y.-J. Ko, E. J. Small, M. Upton, B. Rajeshkumar, and S. P. Balk Selection for Androgen Receptor Mutations in Prostate Cancers Treated with Androgen Antagonist Cancer Res., June 1, 1999; 59(11): 2511 - 2515. [Abstract] [Full Text] [PDF]
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C. W. Gregory, D. Kim, P. Ye, A. J. D’Ercole, T. G. Pretlow, J. L. Mohler, and F. S. French Androgen Receptor Up-Regulates Insulin-Like Growth Factor Binding Protein-5 (IGFBP-5) Expression in a Human Prostate Cancer Xenograft Endocrinology, May 1, 1999; 140(5): 2372 - 2381. [Abstract] [Full Text]
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C. Lopez-Otin and E. P. Diamandis Breast and Prostate Cancer: An Analysis of Common Epidemiological, Genetic, and Biochemical Features Endocr. Rev., August 1, 1998; 19(4): 365 - 396. [Abstract] [Full Text] [PDF]
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N. Poujol, J.-M. Wurtz, B. Tahiri, S. Lumbroso, J.-C. Nicolas, D. Moras, and C. Sultan Specific Recognition of Androgens by Their Nuclear Receptor. A STRUCTURE-FUNCTION STUDY J. Biol. Chem., July 28, 2000; 275(31): 24022 - 24031. [Abstract] [Full Text] [PDF]
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