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Medical Research Council Group in Molecular Endocrinology, CHUL Research Center and Laval University, Québec, Canada G1V 4G2
Address all correspondence and requests for reprints to: Dr. Van Luu-The, Medical Research Council Group in Molecular Endocrinology, CHUL Research Center, 2705 Laurier Boulevard, Québec, Québec, Canada G1V 4G2.
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Abstract |
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 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
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HSD. Using
intact transfected cells, type 5 17ßHSD shows a substrate specificity
pattern comparable to those of human type 3 17ßHSD and mouse type 5
17ßHSD. These enzymes catalyze more efficiently the transformation of
androstenedione (4-dione) to testosterone, whereas the transformation
of dihydrotestosterone to 5-androstane-3,17ß-diol is much
lower. In contrast, type 3 3HSD catalyzes more efficiently the
transformation of dihydrotestosterone to
5-androstane-3,17ß-diol, whereas the transformation of 4-dione
to testosterone represents only 7% of the 3HSD activity. However,
upon homogenization, human type 5 17ßHSD activity decreases to
approximately 10% of the activity in intact cells and remains stable
at this level together with the 3HSD activity. Under the same
conditions, however, the mouse enzyme is not altered by homogenization.
Indeed, using purified human 17ßHSD overexpressed in
Escherichia coli, we could confirm that a much greater
amount of protein is required to produce activity similar to the
enzymatic activity measured in intact transfected cells. The present
data provide the answer to the question of why previous researchers
could hardly detect type 5 17ßHSD activity. Indeed, all previous
publications used cell or tissue homogenates or purified enzymes. Under
such conditions, only the low level, but stable, 3HSD and 17ßHSD
activities could be measured, whereas the high level, but highly
unstable, 17ßHSD activity could not be measured. As type 5 17ßHSD
shares 84%, 86%, and 88% amino acid identity with types 1 and 3
3HSD and 20HSDs, respectively, Northern blot analysis used in
previous studies could not provide unequivocal information. In this
report, we used a more specific ribonuclease protection assay and could
thus show that human type 5 17ßHSD is expressed in the liver,
adrenal, and prostate; in prostatic cancer cell lines DU-145 and LNCaP;
as well as in bone carcinoma (MG-63) cells. By analogy with type 3
17ßHSD, which is responsible for the formation of androgens in the
testis, the expression of type 5 17ßHSD in the prostate and bone
cells suggests that this enzyme is involved in the formation of active
intracellular androgens in these tissues. |
Introduction |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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-androstane-3,17-dione (A-dione) and their respective
backward reactions are catalyzed by 17ß-hydroxysteroid dehydrogenases
(17ßHSDs). This activity is an obligatory step in the biosynthesis of
all androgens and estrogens, namely E2, 5-diol, T, and DHT.
These hormones bind much strongly to their respective receptors in the
17ß-hydroxy configuration than the corresponding 17-keto derivatives.
Impairment of type 3 17ßHSD activity leads to male
pseudohermaphroditism (1) with the absence of the internal male
reproductive structures (epididymis, seminal vesicles, and vas
deferens) that are normally formed from Wolffian ducts under the
influence of testosterone. In the human, five types of 17ßHSDs have been identified, and their structures elucidated. Type 1 17ßHSD (2, 3), originally cloned from human placenta libraries, was also found in the ovaries and mammary glands (4, 5). The enzyme was overproduced in mammalian cells (6) and in a baculovirus system (7) and was found to be functional in a homodimeric form. Its three-dimensional structure has also been recently elucidated (8). Type 2 17ßHSD (9), first isolated from prostate and placenta complementary DNA (cDNA) libraries, has been described in the placenta, liver, small intestine, endometrium, kidney, pancreas, and colon (10). Type 3 17ßHSD, on the other hand, was detected only in the testis and is indeed the gene responsible for pseudohermaphroditism (11). More recently, human type 4 17ßHSD (12), homologous to porcine endometrial 17ßHSD (13), has also been identified.
Using cells transfected with various cDNAs, it has been shown that
these enzymes are substrate and orientation selective (14).
E1 is the preferred substrate for type 1 17ßHSD (6, 14),
whereas T and E2 (9, 14) are the preferred substrates for
type 2. Types 3 (11, 14) and 5 (15) 17ßHSDs catalyze selectively the
transformation of 4-dione to T. Type 4 (12) possesses a relatively weak
activity, catalyzing the transformation of E2 and
DHEA to E1 and 5-diol, respectively. The
corresponding mouse types 1 (16), 2 (17), 3 (18), 4 (19), and 5 (20)
17ßHSD cDNA clones have been isolated and are named according to
their homology with the human clones. Recently, a rat type 6 17ßHSD
was (21) isolated. It possesses high homology with retinol
dehydrogenases and catalyzes selectively the transformation of DHT and
3-diol to A-dione and androsterone (ADT), respectively. A
mouse and rat type 7 17ßHSD were also described (22). This enzyme
catalyzes selectively the transformation of E1 to
E2 and probably plays a major role in the development of
female organs such as the ovary and mammary glands. In this report,
using intact transfected cells as well as a purified enzyme
overexpressed in Escherichia coli, we describe the
characteristics of the highly unstable type 5 17ßHSD. Comparisons
with the characteristics of human type 3 17ßHSD (11), mouse type 5
17ßHSD (20), and human type 3 3HSD (23) are also presented.
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Materials and Methods |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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gt11 cDNA library (Clontech, Palo Alto, CA) using PCR
and an oligoprimer pair
(5'-GGA-AAT-CGT-GAC-AGG-GAA-TGG-ATT-CCA-AAC-AG-3' and
5'-GGA-ATT-CTT-TAT-TGT-ATT-TCT-GGC-CTA-TGG-AGT-GAG-3') was used as
probe to screen the same library to obtain a more complete and reliable
clone, as previously described (25). Positive recombinant phage plaques
were purified, and phage DNA was isolated by centrifugation for 90 min
at 105,000 x g followed by phenol extraction. DNA
inserts were obtained by digestion with EcoRI and subcloned
into a pCMV-neo expression vector. Plasmid DNA was prepared using the
Qiagen Mega kit (Qiagen, Chatsworth, CA). Similarly, the cDNA fragment
of the entire coding region of mouse type 5 17ßHSD (20) was amplified
from mouse liver cDNA library (Clontech) using synthetic
oligonucleotide primers
(5'-GGA-ATT-CAT-GGA-TTC-TAA-GCA-GCA-GAC-AGT-GCG-T-3' and
5'-GGA-ATT-CGC-TCA-GAA-GCT-TGG-ATT-AGG-GTG-3'). The resulting fragment
was subcloned into a pCMVneo expression vector to generate stably
transfected 293 cells as described below.
Stable expression in transformed human embryonal kidney (293)
cells
Cells were cultured in six-well falcon flasks to approximately
3 x 105 cells/well in DMEM (Life Technologies, Burlington, Canada) supplemented with 10%
(vol/vol) FCS (HyClone Laboratories, Inc., Logan, UT) at
37 C under a 95% air-5% CO2 humidified atmosphere. Five
micrograms of pCMVneo-h17ßHSD5 or pCMVneo-m17ßHSD5 plasmids were
transfected using a lipofectin transfection kit (Life Technologies). After 6-h incubation at 37 C, the transfection
medium was removed, and 2 ml DMEM were added. Cells were further
cultured for 48 h and then transferred into 10-cm petri dishes and
cultured in DMEM containing 700 µg/ml G-418 to inhibit the growth of
nontransfected cells. Medium containing G-418 was changed every 2 days
until resistant colonies were observed.
Site-directed mutagenesis and transient transfection
The change of K to E and M to I at positions 75 and 175,
respectively, in human type 5 17ßHSD were performed using
oligonucleotide primer pairs (5'-CAG-ATG-GCA-GTG-TGG-AGA-GAG-AAG-A-3'
and 5'-GTC-TTC-TCT-CTC-CAC-ACT-GCC-ATC-TG-3', and
5'-GGC-AGC-TGG-AGA-TCA-TCC-TCA-ACA-CG-3' and
5'-CTT-GTT-GAG-GAT-GAT-CTC-CAG-CTG-CC-3') and the Quick Change
site-directed mutagenesis kit (Stratagene, La Jolla, CA).
Experiment procedures were carried out as described by the
manufacturer’s instructions. The integrity of the construct was
checked by sequencing of the inserted DNA fragment using the T7
sequencing kit (Pharmacia LKB Biotechnology, Québec,
Canada). Plasmid DNA was prepared using the Qiagen Mega Kit
(Qiagen). Oligonucleotide primers were synthesized with a DNA
synthesizer ABI-394 (Perkin-Elmer/Cetus, Emeryville,
CA)
Transient transfection of the expression vectors into 293 cells was performed using the calcium phosphate procedure (26). pCMV-galactosidase plasmid (0.5 µg) was cotransfected along with the vector expressing type 5 17ßHSD to monitor the efficiency of the transfection. ß-Galactosidase activity was determined using the Galacto-Light Plus Kit (Tropix, Bedford, MA) as described by the manufacturer.
Expression and purification of human type 5 17ßHSD in E.
coli
The coding region of human type 5 17ßHSD was amplified by PCR
using an oligoprimer pair
(5'-GGG-GAA-TTC-ATG-GAT-TCC-AAA-CAG-CAG-TGT-G-3' and
5'-GCC-GGA-ATT-CGG-TGC-TCA-TCA-CCG-GCT-GTT-3') to create an
EcoRI site that allowed the insertion of the fragment in the
EcoRI site of pGEX-1T (Amersham Pharmacia Biotech, Inc., Québec, Canada) and production of a fusion
protein with glutathione-S-transferase. Overexpression of
the type 5 17ßHSD in E. coli, purification of the fusion
protein on the glutathione-Sepharose 4B affinity column, and cleavage
of the fusion protein by thrombin were performed as described by the
manufacturer.
Assay of enzymatic activity
Determination of the activities were performed in intact cells
as previously described (25). Briefly, 0.1 µM of the
14C-labeled steroid indicated (DuPont, Inc., Missisauga,
Canada) was added to freshly changed culture medium in a 24-well
culture plate. When cell homogenates or purified enzyme are used, the
reaction was performed in a final volume of 1 ml 50 mM
sodium phosphate buffer (pH 7.5), 20% glycerol, 1 mM EDTA,
0.1 µM of the 14C-labeled steroid, and 1
mM of the corresponding cofactor (NADPH for reduction and
NADP+ for oxidation). After incubation, the steroids were
extracted twice with 1 ml ether. The organic phases were pooled and
evaporated to dryness. The steroids were solubilized in 50 µl
dichloromethane, applied to Silica Gel 60 TLC plates (Merck, Darmstadt,
Germany) before separation by migration in the toluene-acetone (4:1)
solvent system. Substrates and metabolites were identified by
comparison with reference steroids, revealed by autoradiography, and
quantified using the PhosphorImager System (Molecular Dynamics, Inc., Sunnyvale, CA).
Antibody raising
The peptide sequence NH2-CHSPMSLKPGEELSPTDE-COOH
located at amino acid positions 117–134 was synthesized and purified
on HPLC by the Eastern Quebec Peptide Services (Quebec, Canada). New
Zealand rabbits received a sc injection of 85 µg peptide solubilized
in 1 ml PBS containing 50% Freund’s complete adjuvant. The animals
were boosted twice with 40 µg peptide in 50% Freund’s incomplete
adjuvant at 1-month intervals. Two weeks after the last injection, the
animals were killed by decapitation, and the blood was collected.
Antiserum was obtained by decantation and separation by centrifugation
and was stored at -80 C.
Immunoblot analysis
Immunoblots were performed as previously described (27).
Briefly, 1.5 x 105 293 cells stably transfected with
type 5 17ßHSD and 1.75 µg enzyme purified as described above were
electrophoresed on a 5–15% SDS-polyacrylamide gel as described by
Laemmli (28). The gel was transferred to the nitrocellulose filters
using a Bio-Rad apparatus (Richmond, CA) for 4 h at 60 V. Blots
were treated three times with 5% fat-free milk in PBS containing 1%
Triton X-100 for 30 min. Antiserum against the peptide of human type 5
17ßHSD obtained as described above was added in a dilution of 1:1000
and incubated at 4 C for 18 h. The blot was then washed three
times with PBS containing 5% fat-free milk and 1% Triton X-100. After
washing, antirabbit IgG horseradish peroxidase-linked antibody (from
donkey) was added at a dilution of 1:10,000 and incubated for 1 h
at room temperature. After three washes with the same buffer, the blot
was revealed with Western Blot Chemiluminescence Reagent Plus (New
England Nuclear Life Science Products, Boston, MA). The blot was
exposed for 60 sec to Hyperfilm (Amersham Life Science, Aylesbury,
UK).
Ribonuclease (RNase) protection assay
A cDNA fragment containing nucleotides from positions 841-1018
downstream from the ATG initiating codon was subcloned into Bluescript
KSII+ vector and used to generate a complementary RNA probe
using T7 RNA polymerase (Pharmacia LKB Biotechnology) and
[32P]UTP. The probe was hybridized with the indicated
amount of RNA from various sources. After digestion with RNase A/T1,
the sample was electrophoresed under denaturing conditions on a 6%
acrylamide gel to detect protected fragments and exposed to Kodak XAR-5
films (Eastman Kodak Co., Rochester, NY) overnight at -80
C with an intensifying screen. Polyadenylated [poly(A)+]
RNA from approximately 50 million cells (LNCaP, DU 145, and MG 63) were
obtained from extraction using Tri-Reagent RNA/DNA/Protein Isolation
Reagent (Molecular Research Center, Inc., Cincinnati, OH)
and Oligotex messenger RNA (mRNA) kits (Qiagen, Missisauga, Canada).
Extraction was performed as indicated by the manufacturer.
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Results |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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HSDHSD, we have compared the 17ßHSD and 3HSD
activities of type 5 human 17ßHSD with those of the well known type 3
17ßHSD (11), the enzyme responsible for the pseudohermaphroditism in
deficient patients, and type 3 3HSD, which we have isolated from a
prostate cDNA library (23). To obtain a more complete picture, we also
compared them with the mouse type 5 17ßHSD (20). As illustrated in
Fig. 1
, cells transfected with human type
5 17ßHSD show a pattern of activity similar to those of human type 3
17ßHSD and mouse type 5 17ßHSD. These enzymes preferentially
transform 4-dione to T, whereas the type 3 3HSD preferentially
transforms DHT to 3-diol. The present data clearly indicate that
human type 5 17ßHSD possesses much higher 17ßHSD activity than
3HSD activity and shows an activity pattern similar to that of mouse
type 5 and human type 3 17ßHSDs.
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HSDHSD shows only a tw-amino acid difference from our type 5
17ßHSD, we determined whether these amino acid changes could affect
the 17ßHSD activity or if these differences are due to polymorphism
or sequencing error. We thus performed site-directed mutagenesis to
change the Lys at position 75 and the Met at position 175 to Glu (K75E)
and Ile (M175I), respectively. Expression vector bearing the double
mutations K75E and M175I was also constructed. As illustrated in Fig. 2, these substitutions did not
significantly affect the 17ßHSD activity.
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, human type 5 17ßHSD activity was
very labile. In fact, upon homogenization of the cells the activity
decreased to a stable level, which was approximately 10% of the level
observed in the intact cell assay. In contrast, mouse type 5 17ßHSD
was very stable (Fig. 4). The activity
remained almost the same as that found in intact cells. The slight
increase in activity could be due to a higher concentration of added
cofactor.
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HSD
catalyzes the reduction of androstanedione and the oxidation 3-diol
with maximum velocities (Vmax) of 11.1 and 4.83
nmol/min·mg, respectively. In a radiometric assay, the
Vmax for the oxidation of 3-diol was 8.45 nmol/min·mg
protein. In our preparation of purified E. coli-expressed
enzyme, the Vmax for the oxidation of 3-diol to ADT and
DHT and the reduction of 4-dione to T were 25 and 30 nmol/min·mg,
respectively (data from Fig. 6). Although our experimental conditions
and those used by Lin et al. (31) are different, especially
in the substrate concentration (0.1 vs. 1.5–40
µM, respectively), and thus the results could not be
compared closely, they indicate that in the purified enzyme
preparation, the oxidative and reductive reactions are performed at a
similar rate. To determine whether the 17ßHSD activity of the enzyme
expressed in E. coli possesses high or low level type 5
17ßHSD activity, we took the amount of intact transfected cells and
purified E. coli-expressed enzyme that produced the same
activity and performed Western blot analysis using an antibody specific
to type 5 17ßHSD to estimate the quantity of enzyme in both systems.
As shown in Fig. 5, the amount of
purified enzyme needed to produce a similar activity was higher for
bacteria-expressed enzyme than for that expressed in intact cells. In
addition, as illustrated in Fig. 6, the
17ßHSD activity in the purified enzyme was at the same level as the
3HSD activity.
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, in addition to the
transformation of 4-dione to T, the mouse type 5 17ßHSD efficiently
catalyzes the transformation of E1 to E2 (60%
of the transformation of 4-dione to T), whereas in the human the
transformation of E1 to E2 by type 5 17ßHSD
represents only 5% of the transformation of 4-dione to T. On the other
hand, the human enzyme possesses a high 20HSD activity that
inactivates progesterone to 20-hydroxyprogesterone,
whereas in the mouse this activity represents only 10% of the 17ßHSD
activity.
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HSD as well as 20HSD (Fig. 8). Northern blot analysis could not
differentiate these different mRNAs. To determine the importance of
type 5 17ßHSD in peripheral tissues, especially in the prostate, we
used a RNase protection assay to detect specifically the mRNA
expression level of type 5 17ßHSD in the liver, adrenal, and
prostate; in the prostatic carcinoma cell lines, DU-145 and LNCaP; and
in osteosarcoma cells, MG-63 (Fig. 9).
Type 5 17ßHSD was found in all of the tissues mentioned above.
However, the level of mRNA expression in the prostate and adrenal was
much lower than that in the liver, prostatic cancer, and osteocarcinoma
cells.
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Discussion |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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HSD (30, 31). Upon homogenization, the enzyme possesses
stable, but very weak, 17ßHSD activity (Ref. 31 and this report). The
level of the remaining 17ßHSD activity is similar to that of the
3HSD activity and is of minor importance compared with
17ßHSD activity measured in intact cells.
In contrast to human type 5 17ßHSD, the mouse enzyme is stable and is
not altered upon homogenization. These two enzymes share 76% amino
acid identity, the percentage generally observed for orthologous
protein between the human and mouse species. It was somewhat expected
that the corresponding mouse and human enzymes could have different
substrate specificity patterns. Indeed, the mouse type 5 17ßHSD shows
higher activity for C18 steroids; this activity was one
third of that for C19 substrates. In the human, the ability
of the enzyme to transform C18 steroids is much weaker.
However, the human enzyme possesses much higher 20HSD activity that
catalyzes the inactivation of progesterone to
20-hydroxyprogesterone. High activity is not always related to a
better physiological relevance, e.g. human type 1 3ßHSD,
which possesses 10-fold higher affinity and higher activity for
pregnenolone and DHEA than type 2 3ßHSD (32), is not
responsible for congenital adrenal hyperplasia (33) due to 3ßHSD
deficiency. This disease was due to the deficit of type 2 3ßHSD,
which exhibits lower activity. On the other hand, for an enzyme that
possesses multiple activities, the chosen activity will depend on the
tissue and cell type where the enzyme is expressed. In the prostate, a
male organ, it is well known that the female hormone
progesterone has little effect, probably due to the low
level of progesterone and the lack of
progesterone receptors. The presence of type 5 17ßHSD in
the prostate strongly suggests its role in the transformation of
4-dione to T, as type 3 17ßHSD, the gene responsible for testicular
17ßHSD deficiency, is not expressed in this organ (34). However, the
possible role of type 5 17ßHSD in better protection of the prostate
from progesterone, a female-specific hormone, could not be
rule out.
As illustrated in Fig. 7
, the transformation of
progesterone by human type 5 17ßHSD is about 2.5-fold
higher than the transformation of 4-dione. Accordingly, time-course
studies at 0, 1, 2, 4, 8, and 24 h using intact transfected cells
show a 2.5-fold higher activity for the transformation of
progesterone (data not shown). It also appears that
17ßHSD and other steroidogenic enzymes in rodents show
different substrate specificity patterns compared to their human
counterparts. Indeed, although human type 1 17ßHSD is selective for
the transformation of E1 to E2, the mouse
enzyme also transforms C19 steroids (16). A similar
situation has been observed for DHEA sulfotransferase.
This enzyme in humans is a unique enzyme that transforms 3- as well
as 3ß-hydroxysteroids and C19 as well as C21
and C27 steroids (35), whereas in the guinea pig, there is
one enzyme that is selective for C19 3-steroids (36),
and the other is more selective for C21 3ß-steroids (37).
On the other hand, in the rat and mouse, there are up to six types of
different hydroxysteroid sulfotransferases (35).
The low level of type 5 17ßHSD mRNA expression in the prostate is probably due to the relatively fewer cells expressing this enzyme compared to the total number of cells in this organ. Indeed, using in situ hybridization and immunocytochemistry, Mohamed et al. (38), showed that type 5 17ßHSD is mainly expressed in the basal epithelial cells of the prostate. These cells only represent a small part of the cells in the prostate, which contains a larger amount of fibroblasts and smooth muscle cells.
Comparison of the mouse type 5 17ßHSD gene with the corresponding
human gene indicates that these two genes have similar structural
organization and size (
16 kb; Rheault, P., et al.,
unpublished results). However, the human type 1 3HSD, which shares
79% identity with human type 5 17ßHSD, has a longer length (20 kb).
Such results suggest that type 5 17ßHSD is an important enzyme that
is conserved between species. Indeed, in contrast to most gene
families, where high sequence identity means similar activity, members
of the human aldo-keto reductase family show high amino acid sequence
identity but possess different activities. A striking example is type 3
3HSD and 20HSD, which differ by only seven amino acids (23). On
the other hand, human type 5 17ßHSD shares 84% and 87% amino acid
identity with human type 1 3HSD and 20HSD, respectively. It seems
that members of this gene family duplicated recently and are in the
process of fixation or elimination. These genes are thought to be
generated by duplication and are located in the same clusters. Indeed,
using cDNA encoding type 1 3HSD as a probe, this cluster was mapped
by fluorescence in situ hybridization to chromosome
10p14–15 (30). Using a DNA fragment from the human type 5 17ßHSD
gene, we have confirmed the localization of type 5 17ßHSD in this
cluster (Dufort, I., et al., unpublished results).
Although purified enzymes are the preferred materials for studying kinetic and binding interactions as well as catalytic properties, the present results strongly suggest that the use of intact transfected cells in culture is more suitable to characterize physiological activity, especially for oxidoreductase enzymes. In this manner, the preferred reaction of the enzyme, oxidation or reduction, is clearly identified, and unstable or labile enzyme can be analyzed with confidence. As an example, using cells stably expressing the mouse type 5 17ßHSD, we could clearly show that the enzyme catalyzes preferentially the transformation of 4-dione and A-dione to T and DHT, respectively. These data are in contrast with a previous report from Deyashi et al. (20), who described this enzyme as estradiol 17ß-dehydrogenase. The discrepancy is probably due to the fact that Deyashi et al. (20) used cell subfractions and the cofactor NAD+, and thus forced the reaction in the oxidative direction, which is not a preferred reaction in intact cells.
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Acknowledgments |
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Footnotes |
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Received July 27, 1998.
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P.-G. Blanchard and V. Luu-The Differential androgen and estrogen substrates specificity in the mouse and primates type 12 17{beta}-hydroxysteroid dehydrogenase J. Endocrinol., August 1, 2007; 194(2): 449 - 455. [Abstract] [Full Text] [PDF]
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W. Qiu, M. Zhou, M. Mazumdar, A. Azzi, D. Ghanmi, V. Luu-The, F. Labrie, and S.-X. Lin Structure-based Inhibitor Design for an Enzyme That Binds Different Steroids: A POTENT INHIBITOR FOR HUMAN TYPE 5 17beta-HYDROXYSTEROID DEHYDROGENASE J. Biol. Chem., March 16, 2007; 282(11): 8368 - 8379. [Abstract] [Full Text] [PDF]
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K. Blouin, C. Richard, G. Brochu, F.-S. Hould, S. Lebel, S. Marceau, S. Biron, V. Luu-The, and A. Tchernof Androgen inactivation and steroid-converting enzyme expression in abdominal adipose tissue in men J. Endocrinol., December 1, 2006; 191(3): 637 - 649. [Abstract] [Full Text] [PDF]
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 D. R. Bauman, S. Steckelbroeck, D. M. Peehl, and T. M. Penning Transcript Profiling of the Androgen Signal in Normal Prostate, Benign Prostatic Hyperplasia, and Prostate Cancer Endocrinology, December 1, 2006; 147(12): 5806 - 5816. [Abstract] [Full Text] [PDF]
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 M. Stanbrough, G. J. Bubley, K. Ross, T. R. Golub, M. A. Rubin, T. M. Penning, P. G. Febbo, and S. P. Balk Increased expression of genes converting adrenal androgens to testosterone in androgen-independent prostate cancer. Cancer Res., March 1, 2006; 66(5): 2815 - 2825. [Abstract] [Full Text] [PDF]
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K. Qin, D. A. Ehrmann, N. Cox, S. Refetoff, and R. L. Rosenfield Identification of a Functional Polymorphism of the Human Type 5 17{beta}-Hydroxysteroid Dehydrogenase Gene Associated with Polycystic Ovary Syndrome J. Clin. Endocrinol. Metab., January 1, 2006; 91(1): 270 - 276. [Abstract] [Full Text] [PDF]
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T. Suzuki, Y. Miki, Y. Nakamura, T. Moriya, K. Ito, N. Ohuchi, and H. Sasano Sex steroid-producing enzymes in human breast cancer Endocr. Relat. Cancer, December 1, 2005; 12(4): 701 - 720. [Abstract] [Full Text] [PDF]
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F Labrie, V Luu-The, A Belanger, S-X Lin, J Simard, G Pelletier, and C Labrie Is dehydroepiandrosterone a hormone? J. Endocrinol., November 1, 2005; 187(2): 169 - 196. [Abstract] [Full Text] [PDF]
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 R Mindnich, F Haller, F Halbach, G Moeller, M H. de Angelis, and J Adamski Androgen metabolism via 17{beta}-hydroxysteroid dehydrogenase type 3 in mammalian and non-mammalian vertebrates: comparison of the human and the zebrafish enzyme J. Mol. Endocrinol., October 1, 2005; 35(2): 305 - 316. [Abstract] [Full Text] [PDF]
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 F. Labrie, A. Belanger, V. Luu-The, C. Labrie, J. Simard, L. Cusan, J. Gomez, and B. Candas Gonadotropin-Releasing Hormone Agonists in the Treatment of Prostate Cancer Endocr. Rev., May 1, 2005; 26(3): 361 - 379. [Abstract] [Full Text] [PDF]
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Y. Nakamura, T. Suzuki, M. Nakabayashi, M. Endoh, K. Sakamoto, Y. Mikami, T. Moriya, A. Ito, S. Takahashi, S. Yamada, et al. In situ androgen producing enzymes in human prostate cancer Endocr. Relat. Cancer, March 1, 2005; 12(1): 101 - 107. [Abstract] [Full Text] [PDF]
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F. Labrie, V. Luu-The, E. Calvo, C. Martel, J. Cloutier, S. Gauthier, P. Belleau, J. Morissette, M.-H. Levesque, and C. Labrie Tetrahydrogestrinone induces a genomic signature typical of a potent anabolic steroid J. Endocrinol., February 1, 2005; 184(2): 427 - 433. [Abstract] [Full Text] [PDF]
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 K. Blouin, S. Blanchette, C. Richard, P. Dupont, V. Luu-The, and A. Tchernof Expression and activity of steroid aldoketoreductases 1C in omental adipose tissue are positive correlates of adiposity in women Am J Physiol Endocrinol Metab, February 1, 2005; 288(2): E398 - E404. [Abstract] [Full Text] [PDF]
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 D. R. Bauman, S. I. Rudnick, L. M. Szewczuk, Y. Jin, S. Gopishetty, and T. M. Penning Development of Nonsteroidal Anti-Inflammatory Drug Analogs and Steroid Carboxylates Selective for Human Aldo-Keto Reductase Isoforms: Potential Antineoplastic Agents That Work Independently of Cyclooxygenase Isozymes Mol. Pharmacol., January 1, 2005; 67(1): 60 - 68. [Abstract] [Full Text] [PDF]
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M. Quinkler, B. Sinha, J. W Tomlinson, I. J Bujalska, P. M Stewart, and W. Arlt Androgen generation in adipose tissue in women with simple obesity - a site-specific role for 17{beta}-hydroxysteroid dehydrogenase type 5 J. Endocrinol., November 1, 2004; 183(2): 331 - 342. [Abstract] [Full Text] [PDF]
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O. O. Oduwole, Y. Li, V. V. Isomaa, A. Mantyniemi, A. E. Pulkka, Y. Soini, and P. T. Vihko 17{beta}-Hydroxysteroid Dehydrogenase Type 1 Is an Independent Prognostic Marker in Breast Cancer Cancer Res., October 15, 2004; 64(20): 7604 - 7609. [Abstract] [Full Text] [PDF]
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N. M. Holmes, W. L. Miller, and L. S. Baskin Lack of Defects in Androgen Production in Children with Hypospadias J. Clin. Endocrinol. Metab., June 1, 2004; 89(6): 2811 - 2816. [Abstract] [Full Text] [PDF]
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S. Steckelbroeck, Y. Jin, S. Gopishetty, B. Oyesanmi, and T. M. Penning Human Cytosolic 3{alpha}-Hydroxysteroid Dehydrogenases of the Aldo-keto Reductase Superfamily Display Significant 3{beta}-Hydroxysteroid Dehydrogenase Activity: IMPLICATIONS FOR STEROID HORMONE METABOLISM AND ACTION J. Biol. Chem., March 12, 2004; 279(11): 10784 - 10795. [Abstract] [Full Text] [PDF]
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K. Blouin, C. Richard, C. Belanger, P. Dupont, M. Daris, P. Laberge, V. Luu-The, and A. Tchernof Local Androgen Inactivation in Abdominal Visceral Adipose Tissue J. Clin. Endocrinol. Metab., December 1, 2003; 88(12): 5944 - 5950. [Abstract] [Full Text] [PDF]
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 T. S. A. Samy, R. Zheng, T. Matsutani, L. W. Rue III, K. I. Bland, and I. H. Chaudry Mechanism for normal splenic T lymphocyte functions in proestrus females after trauma: enhanced local synthesis of 17{beta}-estradiol Am J Physiol Cell Physiol, July 1, 2003; 285(1): C139 - C149. [Abstract] [Full Text] [PDF]
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M. Quinkler, C. Bumke-Vogt, B. Meyer, V. Bahr, W. Oelkers, and S. Diederich The Human Kidney Is a Progesterone-Metabolizing and Androgen-Producing Organ J. Clin. Endocrinol. Metab., June 1, 2003; 88(6): 2803 - 2809. [Abstract] [Full Text] [PDF]
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F. Labrie, V. Luu-The, C. Labrie, A. Belanger, J. Simard, S.-X. Lin, and G. Pelletier Endocrine and Intracrine Sources of Androgens in Women: Inhibition of Breast Cancer and Other Roles of Androgens and Their Precursor Dehydroepiandrosterone Endocr. Rev., April 1, 2003; 24(2): 152 - 182. [Abstract] [Full Text] [PDF]
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P. Harkonen, S. Torn, R. Kurkela, K. Porvari, A. Pulkka, A. Lindfors, V. Isomaa, and P. Vihko Sex Hormone Metabolism in Prostate Cancer Cells during Transition to an Androgen-Independent State J. Clin. Endocrinol. Metab., February 1, 2003; 88(2): 705 - 712. [Abstract] [Full Text] [PDF]
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V. Nahoum, A. Gangloff, P. Legrand, D.-W. Zhu, L. Cantin, B. S. Zhorov, V. Luu-The, F. Labrie, R. Breton, and S.-X. Lin Structure of the Human 3alpha -Hydroxysteroid Dehydrogenase Type 3 in Complex with Testosterone and NADP at 1.25-A Resolution J. Biol. Chem., November 2, 2001; 276(45): 42091 - 42098. [Abstract] [Full Text] [PDF]
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M. K. Gupta, D. H. Geller, and R. J. Auchus Pitfalls in Characterizing P450c17 Mutations Associated with Isolated 17,20-Lyase Deficiency J. Clin. Endocrinol. Metab., September 1, 2001; 86(9): 4416 - 4423. [Abstract] [Full Text] [PDF]
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T. S. A. Samy, M. W. Knoferl, R. Zheng, M. G. Schwacha, K. I. Bland, and I. H. Chaudry Divergent Immune Responses in Male and Female Mice after Trauma-Hemorrhage: Dimorphic Alterations in T Lymphocyte Steroidogenic Enzyme Activities Endocrinology, August 1, 2001; 142(8): 3519 - 3529. [Abstract] [Full Text] [PDF]
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T. Suzuki, A. D. Darnel, J.-I. Akahira, N. Ariga, S. Ogawa, C. Kaneko, J. Takeyama, T. Moriya, and H. Sasano 5{{alpha}}-Reductases in Human Breast Carcinoma: Possible Modulator of in Situ Androgenic Actions J. Clin. Endocrinol. Metab., May 1, 2001; 86(5): 2250 - 2257. [Abstract] [Full Text]
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S. Steckelbroeck, M. Watzka, R. Reichelt, V. H. J. Hans, B. Stoffel-Wagner, D. D. Heidrich, J. Schramm, F. Bidlingmaier, and D. Klingmüller Characterization of the 5{{alpha}}-Reductase-3{{alpha}}-Hydroxysteroid Dehydrogenase Complex in the Human Brain J. Clin. Endocrinol. Metab., March 1, 2001; 86(3): 1324 - 1331. [Abstract] [Full Text]
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I. Dufort, F. Labrie, and V. Luu-The Human Types 1 and 3 3{{alpha}}-Hydroxysteroid Dehydrogenases: Differential Lability and Tissue Distribution J. Clin. Endocrinol. Metab., February 1, 2001; 86(2): 841 - 846. [Abstract] [Full Text]
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B. Lin, J. T. White, C. Ferguson, S. Wang, R. Vessella, R. Bumgarner, L. D. True, L. Hood, and P. S. Nelson Prostate Short-Chain Dehydrogenase Reductase 1 (PSDR1): A New Member of the Short-Chain Steroid Dehydrogenase/Reductase Family Highly Expressed in Normal and Neoplastic Prostate Epithelium Cancer Res., February 1, 2001; 61(4): 1611 - 1618. [Abstract] [Full Text]
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P. R. Provost, C. H. Blomquist, C. Godin, X.-F. Huang, N. Flamand, V. Luu-The, D. Nadeau, and Y. Tremblay Androgen Formation and Metabolism in the Pulmonary Epithelial Cell Line A549: Expression of 17{beta}-Hydroxysteroid Dehydrogenase Type 5 and 3{alpha}-Hydroxysteroid Dehydrogenase Type 3 Endocrinology, August 1, 2000; 141(8): 2786 - 2794. [Abstract] [Full Text] [PDF]
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P. J. O'Shaughnessy, P. J. Baker, M. Heikkila, S. Vainio, and A. P. McMahon Localization of 17{beta}-Hydroxysteroid Dehydrogenase/17-Ketosteroid Reductase Isoform Expression in the Developing Mouse Testis--Androstenedione Is the Major Androgen Secreted by Fetal/Neonatal Leydig Cells Endocrinology, July 1, 2000; 141(7): 2631 - 2637. [Abstract] [Full Text] [PDF]
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 P. A. Thompson and C. Ambrosone Chapter 7: Molecular Epidemiology of Genetic Polymorphisms in Estrogen Metabolizing Enzymes in Human Breast Cancer J Natl Cancer Inst Monographs, July 1, 2000; 2000(27): 125 - 134. [Abstract] [Full Text] [PDF]
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 K.-n. Qin and R. L. Rosenfield Expression of 17{beta}-Hydroxysteroid Dehydrogenase Type 5 in Human Ovary: A Pilot Study Reproductive Sciences, January 1, 2000; 7(1): 61 - 64. [Abstract] [PDF]
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New Insight into the Molecular Basis of 3{beta}-Hydroxysteroid Dehydrogenase Deficiency: Identification of Eight Mutations in the HSD3B2 Gene in Eleven Patients from Seven New Families and Comparison of the Functional Properties of Twenty-Five Mutant Enzymes J. Clin. Endocrinol. Metab., December 1, 1999; 84(12): 4410 - 4425. [Abstract] [Full Text]
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S. Gingras and J. Simard Induction of 3{beta}-Hydroxysteroid Dehydrogenase/ Isomerase Type 1 Expression by Interleukin-4 in Human Normal Prostate Epithelial Cells, Immortalized Keratinocytes, Colon, and Cervix Cancer Cell Lines Endocrinology, October 1, 1999; 140(10): 4573 - 4584. [Abstract] [Full Text]
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M. A. English, K. F. Kane, N. Cruickshank, M. J. S. Langman, P. M. Stewart, and M. Hewison Loss of Estrogen Inactivation in Colonic Cancer J. Clin. Endocrinol. Metab., June 1, 1999; 84(6): 2080 - 2085. [Abstract] [Full Text]
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X.-F. Huang and V. Luu-The Molecular Characterization of a First Human 3(alpha right-arrowbeta )-Hydroxysteroid Epimerase J. Biol. Chem., September 15, 2000; 275(38): 29452 - 29457. [Abstract] [Full Text] [PDF]
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R. Zheng, T. S. A. Samy, C. P. Schneider, L. W. Rue III, K. I. Bland, and I. H. Chaudry Decreased 5alpha -dihydrotestosterone catabolism suppresses T lymphocyte functions in males after trauma-hemorrhage Am J Physiol Cell Physiol, June 1, 2002; 282(6): C1332 - C1338. [Abstract] [Full Text] [PDF]
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