(Circulation. 1999;99:2688-2693.)
© 1999 American Heart Association, Inc.
Basic Science Reports |
Variant Estrogen and Progesterone Receptor Messages in Human Vascular Smooth Muscle
From the Department of Medicine, Division of Cardiology (Y.K.H., L.D.H.) and Division of Endocrinology (J.K.R., K.B.H.), University of Colorado Health Sciences Center, Denver, Colo.
Correspondence to Lawrence D. Horwitz, MD, Cardiology B130, University of Colorado Health Sciences Center, Denver, CO 80262. E-mail lawrence.horwitz{at}uchsc.edu
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Abstract |
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Background—Estrogens stimulate growth of breast or uterine cells but have the opposite effect on vascular smooth muscle cells, in which they protect against coronary artery disease with or without concomitant administration of progesterone. A possible cause of differences in hormone action is variable tissue-specific expression of hormone receptor. Therefore, we analyzed the structure of estrogen receptors (ERs) and progesterone receptors (PRs) in human vascular smooth muscle.
Methods and Results—RNA was isolated from human vascular smooth
muscle, and the functional domains of ER-
and PR were characterized
by reverse transcriptase and polymerase chain reaction. Interestingly,
in addition to wild-type ER- and PR, 5 variant ER- and 2 variant
PR transcripts were found. These variants contained precise deletions
of exons encoding regions of the hormone-binding domain. The PR
transcripts lacked exon 4 (PR
4) and exon 6 (PR6). The ER-
transcripts were missing exon 4 (ER4), exon 5 (ER5), exon 6
(ER6), exon 7 (ER7), and exons 6 and 7, (ER6,7). ER-ß
variants were also detected. The PR variants were functionally
characterized, and PR6 was found to be a dominant-negative
transcription inhibitor of wild-type receptors. Variant PR
was present in premenopausal women but absent in postmenopausal
women.
Conclusions—Variant PR and ER transcripts are extensively expressed in human vascular smooth muscle. The complex tissue-specific effects of sex hormones may be mediated by the expression of heterogeneous forms of their cognate receptors. The presence of variant ERs and PRs may be of importance in altering the physiological effects of estrogens or progestins in vascular smooth muscle.
Key Words: genes • muscle, smooth • receptors • hormones • coronary disease
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Introduction |
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TopAbstractIntroductionMethodsResultsDiscussionReferences |
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The incidence of coronary artery disease in postmenopausal women is markedly reduced by treatment with exogenous estrogens.1 A possible mechanism of this effect is inhibition by estrogens of vascular smooth muscle cell growth and migration, a process critical to atherogenesis and vascular restenosis.2 3 4 Estrogen receptors (ERs) are ligand-activated transcription factors. Activated ER binds to estrogen response elements (EREs) in the promoters of target genes, thereby regulating gene expression.5 Among the proteins regulated by estrogens in vascular smooth muscle cells are the progesterone receptors (PRs).6
The effects of estrogens vary markedly in different tissues; for
example, estrogens stimulate growth of breast and uterine
cells7 8 but inhibit growth of vascular smooth muscle
cells.2 3 4 Progesterone exhibits antagonism to estrogen
effects in the uterus9 10 but enhances the effects of
estrogens in vascular smooth muscle.11 12 A possible
explanation for the heterogeneity of estrogen or
progesterone action in different targets is that some tissues express
variant forms of ER or PR that have transcriptional effects different
from the "wild-type" receptors. There are 2 wild-type ERs,
designated ER- and ER-ß,13 14 and 2 wild-type PR
isoforms, called PR-A and PR-B.15 16 Additionally, mRNA
exon deletion splice variants may arise that encode altered receptor
proteins that are missing key functional domains. Variant forms of
ER- arising from exon splicing errors have been observed in breast
cancers, meningiomas, and normal human mammary
tissue.17 18 19 Recently, Inoue et al20
reported 3 exon deletion variants of ER in vascular smooth muscle cells
isolated from rat aorta. PR splice variants have been reported in
breast cancer.21 22
Variant receptors are capable of anomalous transcriptional activities that may dominantly inhibit or enhance the effects of wild-type receptors.23 24 In addition, some variant receptors are constitutively active: they can mimic the transcriptional effects of estrogens or progestins in the absence of hormone. Finally, some variant receptor messages are likely to be incapable of being translated into protein. If these are the most prevalent types of receptor mRNA present, normal receptor-mediated actions of estrogens or progestins may be absent. Therefore, expression of variant ER or PR in a particular tissue, such as vascular smooth muscle, could have effects that differ markedly from those of the wild-type receptors.
To analyze the expression patterns of wild-type and variant ER
and PR mRNAs, we used reverse transcriptase (RT) and the polymerase
chain reaction (PCR) in human vascular smooth muscle. We identified
several ER- and ER-ß variants. We also found 2 PR variants that we
cloned and sequenced to identify the exon deletions. One of the PR
variants, an exon 6 deletion mutant, is a dominant-negative
inhibitor.21 We propose that the complex,
tissue-specific effects of sex hormones may be mediated in part by the
expression, or lack thereof, of heterogeneous types of
their cognate receptors.
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Methods |
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TopAbstractIntroductionMethodsResultsDiscussionReferences |
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Tissue Culture
As outlined in the Table
, normal
and diseased coronary artery, saphenous vein, aorta, mammary
artery, and iliac artery were used. The sections were placed into
DMEM with 1% (vol/vol) antibiotic-antimycotic solution at pH
7.4 (DMEM+AA) (Sigma Chemical Co). The fat and connective tissue were
removed, each segment was cut longitudinally, and the
endothelial layer was mechanically denuded. After a
10-minute incubation in Ca2+- and
Mg2+-free HBSS (Gibco BRL) containing 73 U/mL
collagenase (Worthington), the inner muscular layer was
carefully stripped from the outer adventitial layer, and the smooth
muscle was incubated in DMEM+AA for 72 hours at 37°C.
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RNA Isolation
A modified version of a single-step method of RNA isolation by
acid guanidine thiocyanate-phenol-chloroform extraction (RNA STAT-60,
Tel-Test B Inc) was used to purify the RNA. RNase-free DNase I was used
to remove DNA contamination from the RNA.
RT-PCR
The GeneAmp RNA PCR kit (Perkin-Elmer) was used with 0.5- to
1.0-µg samples of total RNA and 2.5 µg of murine leukemia virus
(MuLV) RT. The RNA was heated to 70°C for 5 minutes before addition
of the RT. The RT reaction was performed at 42°C for 60 minutes, then
heated to 99°C for 5 minutes to destroy the enzyme. PCR was performed
with primers specific for the receptor domains of interest. An initial
cycle at 94°C for 1 minute was followed by 35 cycles at 94°C for 30
seconds, 60°C for 40 seconds, 72°C for 90 seconds, and a final
extension cycle at 72°C for 7 minutes.
[32P]-dCTP (DuPont NEN) was incorporated
into the PCR reaction. The products were resolved on 1.5%
agarose gels.
Cloning
The PCR products were inserted into a plasmid vector with a
TA cloning kit (Invitrogen) immediately after the reamplified PCR
products were obtained. The ligation reaction was performed
overnight at 15°C. Cloned amplified PCR underwent automated
sequencing at Biotechnology Resource Center, Cornell
University.
Primers
Oligonucleotide primers were designed for PCR
amplification of specific DNA fragments of ER and PR. A primer for
ß-actin served as an RNA quantification control. The primers
amplified specific exons in the DNA binding domain (DBD) or hormone
binding domain (HBD) of ER- (exons 1 to 4) and PR (exons 5 to 8).
These primers were able to detect deletions or amplifications in exons
2 to 7 for both ER- and PR. The primer locations were as follows:
primers 1 and 2 (nucleotide [nt] 617 to 637, nt 1082 to
1103) for ER- DBD (487 bp); primers 3 and 4 (nt 974 to 994, nt 1787
to 1811) for ER- HBD (840 bp); primers 5 and 6 (nt 2355 to 2375, nt
2675 to 2697) for PR DBD (345 bp); and primers 7 and 8 (nt 2619 to
2642, nt 3393 to 3411) for PR HBD (824 bp).
The primers used for ER-ß amplified both the DBD and HBD (nt 174 to 303, nt 1354 to 1383), generating a fragment of 1212 bp. All the primers were tested on ER and PR cDNA plasmid clones to ensure that they generated the specific products targeted.
Construction of Exon Deletion Mutants
The cDNAs encoding the PR variants were created by PCR with
pSG5-hPR1 or -hPR2 (expression vectors for the B or A isoforms of PR)
as templates and oligonucleotides designed to
selectively eliminate the desired exon. The validity of the constructs
was confirmed by sequencing.
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Results |
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Identification of ER-
and PR Transcripts in Human Vascular
Smooth Muscle by RT-PCRTo analyze the ER-
and PR transcripts, primers were
designed to amplify the encoding mRNA segments of the DBD or HBD of
these receptors. RT-PCR was performed with each primer pair, and the
labeled products were resolved on an agarose gel. A
representative result from smooth muscle tissue
isolated from the saphenous vein of a premenopausal woman is shown in
Figure 1. The black arrow in each lane
marks the predicted wild-type DBD or HBD band for each receptor; all
other bands are due to variant transcripts. Note the presence of 
2
variants in lanes 2 and 3 and 3 variants in lane 4. The variant bands
marked by white arrows were isolated, cloned, and sequenced.
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The mRNAs of all steroid receptors, including ER- and PR, are
assembled from 8 exons. Exon 1 encodes the N-terminus; exons 2 and 3
the DBD; exon 4 the nuclear location signal (NLS), hinge, and proximal
region of the HBD; and exons 5 to 8 the rest of the HBD. The predicted
protein structure of the variant ER-, determined from mRNA extracted
from saphenous vein vascular smooth muscle cells, is shown in Figure 2 for ER- compared with the wild-type
receptors. Variant ER- mRNAs present in vascular smooth muscle
cells include deletions of exon 4 (4), exon 5 (5), exon 6 (6),
exon 7 (7), and exons 6 and 7 (6,7) (Figure 2). Three of
these deletions (5, 6, and 7) produce a frameshift generating
a missense protein (black boxes) with an early termination codon,
yielding truncated receptors. Two others, the 4 and 6,7
deletions, do not generate frameshifts. Instead, these ER- variants
do not encode some amino acids in the proximal region (4) or central
region (6,7) of the HBD.
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We identified 2 variant PR mRNAs in vascular smooth muscle cells. These
were cloned and sequenced (Figure 3). One
is an exon 4 deletion (4). This modification removes the NLS, hinge,
and proximal HBD from the receptors, but because no frameshift occurs,
the remainder of the HBD would be translated normally. The second PR
variant is an exon 6 (6) deletion. This produces a frameshift
followed by the synthesis of 12 missense amino acids, ending in a
termination codon, yielding a receptor truncated in the middle of the
HBD.
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Prevalence of PR Variants in Human Vascular Smooth Muscle
Results of studies of PR variants in 15 subjects are shown in the
Table
. The subjects included 5 premenopausal women, 4
postmenopausal women, and 6 men. The men ranged in age from 19 to 69
years. The samples obtained from men contained widely varying
percentages of variant or wild-type PR. PR was present in all the
samples from premenopausal women, but a substantial percentage of the
PR was of the variant type. The samples from the 4 postmenopausal women
contained wild-type but very little or no variant PR. Therefore, in
this small sampling of subjects, variant PR appears to be very
prevalent in premenopausal women but essentially absent in
postmenopausal women.
ER-ß Variant Transcripts Are Present in Human Vascular
Smooth Muscle
In Figure 4, the vascular smooth
muscle in a saphenous vein segment removed from a premenopausal woman
expresses variant ER-ß transcripts. In addition to the wild-type
ER-ß bands (black arrows), several variant PCR products (open
arrows) are present. We found 1 variant band in the HBD that was
larger than the wild type. A similar variant was described recently in
rat ovary and prostate cells.25
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Discussion |
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TopAbstractIntroductionMethodsResultsDiscussionReferences |
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Although premenopausal women and postmenopausal women treated with estrogens have lower incidences of coronary artery disease than men or postmenopausal women not receiving hormone therapy,1 the genomic mechanisms by which this protection is afforded are unknown. Estrogens probably influence atherogenesis and vascular injury through regulation of gene transcription in vascular smooth muscle or endothelium. Estrogens modulate gene transcription by binding to the HBD of ERs.5 The activated DBD of the receptor then binds to the ERE sequences of targeted genes, regulating their transcription. There are 2 wild-type ERs: ER-
, which plays an important role in breast and
uterine cells, and ER-ß, which has a different tissue expression
pattern.13 14 Wild-type PRs are larger proteins than ERs
but have the same general structural organization as ERs, with 1
additional complexity: there are 2 natural isoforms, the A and B
receptors.15 16 There is functional ER in vascular smooth muscle,6 26 and there is evidence in cell culture and animal models that estrogens inhibit vascular smooth muscle cell growth and migration.2 3 4 Interestingly, this growth-inhibitory effect of estrogens in vascular smooth muscle differs from the action of estrogens in other tissues, particularly breast and uterus, where estrogens enhance cell proliferation.7 8
Progestins oppose the growth-stimulatory effects of estrogens in uterus and enhance cell proliferation in breast.9 10 In 2 recent studies,11 12 a progestin in high, probably nonphysiological, dosages had growth-inhibitory effects similar to those of estrogens in cultured human and rat vascular smooth muscle. However, in a study in atherosclerotic monkeys,27 a progestin, medroxyprogesterone (MPA), attenuated estrogen-induced coronary vasodilation. In a study in normal monkeys, MPA prevented inhibition of coronary vasospasm by estradiol, but progesterone did not.28 Additional evidence of possible adverse vascular effects of MPA is provided by a recent clinical trial in which use of high-dose MPA in conjunction with estrogen in postmenopausal women with preexisting coronary artery disease increased the risk of adverse cardiac events during the first year.29 Thus, the interactions of progestins and estrogens in vascular smooth muscle are complex and poorly understood. In different tissues, there is marked heterogeneity of estrogen action and, probably, variation in the interactions between estrogens and progestins. One possible explanation for these diverse physiological effects is that the forms of the receptor proteins expressed differ from organ to organ.
Variant ERs arising from exon splicing errors have been reported in
human breast cancers and normal human mammary
tissue.17 18 19 Recently, Inoue et al20
reported the presence of 3 ER- variants in smooth muscle isolated
from rat aorta. Variant receptors may have anomalous transcriptional
activities: some may dominantly inhibit or enhance the effects of
wild-type receptors. In addition, some variant receptors are
constitutively expressed and may not require binding of their hormone
ligand to produce their effects. Therefore, understanding the
structural types and functional characteristics of variant ERs and PRs
in the vascular bed could lead to valuable insights into the actions of
estrogens and progestins in these tissues. We examined human
vascular smooth muscle to determine whether messages encoding ER-
variants were present.
Using RT-PCR, we discovered the presence of 5 HBD variants of ER- in
vascular smooth muscle: 4, 5, 6, 7, and 6,7 exon
deletions. The 4 variant is not transcriptionally active alone, nor
does it alter the transcriptional activity of wild-type
ER-.19 The 7 variant is not transcriptionally active
alone and does not alter wild-type ER- transcription in HeLa cells,
but it exerts a dominant-negative effect on wild-type ER-
transcription in yeast cells.24 The transcriptional
effects of the 6 and 6,7 ER- variants are unknown. Three of
these transcripts, the 5, 6, and 7 exon deletions, produce
frameshifts that encode missense amino acids, followed by premature
truncation. The other 2, the 4 and 6,7, are without frameshifts.
The 5, 6, 7, and 6,7 variants lack portions of the HBD and
would be expected to be constitutively active in the absence of
hormone; this has been confirmed for the 5
variant.17
Of considerable interest is our finding of PR splice variants in human
vascular smooth muscle. These included a 4 and a 6 exon deletion
variant. Because this is the first description of PR splice variants in
any vascular tissue, we sequenced the mRNA. The 4 deletion removes
the NLS, the hinge, and a portion of the HBD. Because no frameshift
occurs, translation of the remainder of the HBD would be expected. The
6 deletion leads to a frameshift, which truncates the protein in the
HBD. Richer et al21 recently reported the functional
characteristics of these PR variants. The 4 has no transcriptional
activity. The lack of an NLS probably renders this variant unable to
enter the nucleus to compete with the activity of the wild-type PR.
However, the 6 variant, when overexpressed, markedly inhibits the
activity of both isoforms of wild-type PR. Therefore, at least 1 PR
variant is capable of dominantly inhibiting the effects of wild-type
PR. Differences in transcriptional effects, which are dependent on the
type of PR expressed, could cause differences in the effects of
progestins in vascular tissue. There is also evidence that PR can alter
the function of ER.30 31
Samples from postmenopausal women showed little or no PR variant mRNA
in the tissue samples (Table). Three of the 4 postmenopausal
women were taking estrogen replacement therapy with no progestin. It is
possible that exogenous estrogen induces expression of a proportionate
wild-type PR level above that found in premenopausal women and men.
Alternatively, postmenopausal women may produce only wild-type mRNA
regardless of the type of hormonal stimulation.
We discovered by RT-PCR that messages encoding several variants of
ER-ß were also present in vascular smooth muscle. Because the
entire structure of wild-type ER-ß is not known, we did not attempt
to functionally characterize these variants. It has recently been
reported that in ER- knockout mice, estrogens inhibit vascular
smooth muscle growth and migration.32 Because ER-ß was
present in the blood vessels of these mice, ER-ß is probably
transcriptionally active in vascular tissue. The concomitant presence
of variants of ER-ß could have important effects on the
transcriptional activity of wild-type ER-ß and ER-.
We conclude that there are exon deletion variants of ER-, ER-ß,
and PR in human vascular smooth muscle. Because these variants may have
entirely different transcriptional effects than the wild-type receptors
from which they are derived, they are potentially of
physiological importance. An attractive concept is
that some of the puzzling differences in estrogen and progestin effects
in different target tissues could be explained by
heterogeneous expression of their cognate receptor
variants. To further address the possible importance of variant ERs or
PRs, specific antibodies need to be developed for each variant to
determine whether and in what quantities the variant mRNAs are
translated.
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Acknowledgments |
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This work was supported by funding from the American Heart Association, Colorado Affiliate, and NIH grants HL-55291, DK-48238, and CA-26869.
Received December 15, 1998; revision received February 8, 1999; accepted February 12, 1999.
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References |
|---|
|
TopAbstractIntroductionMethodsResultsDiscussionReferences |
|---|
1. Stampfer MJ, Colditz GA. Estrogen replacement therapy and coronary heart disease: a quantitative assessment of the epidemiologic evidence. Prev Med. 1991;20:47–63.[Medline] [Order article via Infotrieve]
2.
Karas RH, Patterson BL, Mendelsohn ME. Inhibition of
vascular smooth muscle cell growth by estrogen. Circulation. 1994;89:1943–1950.
3.
Chen S-J, Li H, Durand J, Oparil S, Chen Y-F. Estrogen
reduces myointimal proliferation after balloon injury of rat carotid
artery. Circulation. 1996;93:577–584.
4. Dai-Do D, Espinosa E, Liu G, Rabelink TJ, Julmy F, Yang Z, Mahler R, Lüscher TF. 17ß-Estradiol inhibits proliferation and migration of human vascular smooth muscle cells: similar effects in cells from postmenopausal females and in males. Cardiovasc Res. 1996;32:980–985.[Medline] [Order article via Infotrieve]
5. Kumar V, Green S, Stack G, Berry M, Jinn J, Chambon P. Functional domains of the human estrogen receptor. Cell. 1987;51:941–945.[Medline] [Order article via Infotrieve]
6. Horwitz KB, Horwitz LD. Canine vascular tissues are targets for androgens, estrogens, progestins, and glucocorticoids. J Clin Invest. 1982;69:750–758.
7. King RJ. A discussion of the roles of oestrogen and progestin in human mammary carcinogenesis. J Steroid Biochem Mol Biol. 1991;39:811–818.[Medline] [Order article via Infotrieve]
8.
Rider V, Carlone DL, Foster RT. Oestrogen and
progesterone control basic fibroblast growth factor mRNA in the rat
uterus. J Endocrinol. 1997;154:75–84.
9. Ordog T, Vertes Z, Vertes M. Role of endogenous opioids in progesterone antagonism on oestradiol-induced DNA synthesis in the rat uterus. J Steroid Biochem Mol Biol. 1993;45:455–457.[Medline] [Order article via Infotrieve]
10. Bhakoo HS, Katzenellenbogen BS. Progesterone antagonism of estradiol-stimulated uterine "induced protein" synthesis. Mol Cell Endocrinol. 1977;8:105–120.[Medline] [Order article via Infotrieve]
11.
Morey AK, Pedram A, Razandi M, Prins BA, Hu R-M,
Biesiada E, Levin ER. Estrogen and progesterone inhibit vascular smooth
muscle proliferation. Endocrinology. 1997;138:3330–3339.
12. Lee W-S, Harder JA, Yoshizumi M, Lee M-E, Haber E. Progesterone inhibits arterial smooth muscle cell proliferation. Nat Med. 1997;3:1005–1008.[Medline] [Order article via Infotrieve]
13.
Kuiper GGJM, Enmark E, Pelto-Huikko M, Nilsson S,
Gustafsson J-A. Cloning of a novel estrogen receptor expressed in rat
prostate and ovary. Proc Natl Acad Sci U S A. 1996;93:5925–5930.
14. Mosselman S, Polman J, Dijkema R. ERß: identification and characterization of a novel human estrogen receptor. FEBS Lett. 1996;392:49–53.[Medline] [Order article via Infotrieve]
15. Kastner P, Krust A, Turotte B, Stropp U, Tora L, Gronemeyer H, Chambon P. Two distinct estrogen-regulated promoters generate transcripts encoding the two functionally different human progesterone receptor forms A and B. EMBO J. 1990;9:1603–1614.[Medline] [Order article via Infotrieve]
16.
Horwitz KB, Alexander PS. In situ photolinked nuclear
progesterone receptors of human breast cancer cells: subunit molecular
weights after transformation and translocation.
Endocrinology. 1983;113:2195–2201.
17.
Fuqua SAW, Fitzgerald SD, Chamness GC, Tandon AK,
McDonnell DP, Nawaz Z, O'Malley BW, McGuire WL. Variant human breast
tumor estrogen receptor with constitutive transcriptional activity.
Cancer Res. 1991;51:105–109.
18.
Leygue ER, Watson PH, Murphy LC. Estrogen receptor
variants in normal human mammary tissue. J Natl Cancer
Inst. 1996;88:284–290.
19. Koehorst SGA, Cox JJ, Donker GH, Lopes de Silva S, Burbach JPH, Thijssen JHH, Blankenstein MA. Functional analysis of an alternatively spliced estrogen receptor lacking exon 4 isolated from MCF-7 breast cancer cells and meningioma tissue. Mol Cell Endocrinol. 1994;101:237–245.[Medline] [Order article via Infotrieve]
20. Inoue S, Hoshino S, Miyoshi H, Akishita M, Hosoi T, Orimo H, Ouchi Y. Identification of a novel isoform of estrogen receptor, a potential inhibitor of estrogen action, in vascular smooth muscle cells. Biochem Biophys Res Commun. 1996;219:766–772.[Medline] [Order article via Infotrieve]
21. Richer JK, Lange CA, Wierman AM, Brooks KM, Tung L, Takimoto GS, Horwitz KB. Progesterone receptor variants found in breast cells repress transcription by wild-type receptor. Breast Cancer Res Treat. 1998;48:231–241.[Medline] [Order article via Infotrieve]
22. Leygue E, Dotzlaw H, Watson PH, Murphy LC. Identification of novel progesterone receptor variant mRNAs in human breast tissue. Biochem Biophys Res Commun. 1996;228:63–68.[Medline] [Order article via Infotrieve]
23.
Wang Y, Miksicek RJ. Identification of a dominant
negative form of the human estrogen receptor. Mol
Endocrinol. 1991;5:1707–1715.
24.
Fuqua SAW, Fitzgerald SD, Allred DC, Elledge RM, Nawaz
Z, McDonnell DP, O'Malley BW, Greene GL, McGuire WL. Inhibition of
estrogen receptor action by a naturally occurring variant in human
breast tumors. Cancer Res. 1992;52:483–486.
25. Chu S, Fuller PJ. Identification of a splice variant of the rat estrogen receptor beta gene. Mol Cell Endocrinol. 1997;132:195–199.[Medline] [Order article via Infotrieve]
26. Karas RH, Patterson BL, Mendelsohn ME. Human vascular smooth muscle cells contain functional estrogen receptor. Circulation. 1994;89:1943–1950.
27. Williams JK, Honore EK, Washburn SA, Clarkson TB. Effects of hormone replacement therapy on reactivity of atherosclerotic coronary arteries in cynomolgus monkeys. J Am Coll Cardiol. 1994;24:1757–1761.[Abstract]
28. Miyagawa K, Roch J, Stanczyk F, Hersmeyer K. Medroxyprogesterone interfered with ovarian steroid protection against coronary vasospasm. Nat Med. 1997;3:324–327.[Medline] [Order article via Infotrieve]
29.
Hulley S, Grady D, Bush T, Furberg C, Herrington D,
Riggs B, Vittinghoff E. Randomized trial of estrogen plus
progestin for secondary prevention of coronary artery disease
in postmenopausal women: Heart and Estrogen/progestin Replacement Study
(HERS) research group. JAMA. 1998;280:605–613.
30.
Takimoto G, Hovland AR, Tasset DM, Melville MT, Tung L,
Horwitz KB. Role of phosphorylation on DNA binding and
transcriptional functions of human progesterone receptors. J
Biol Chem. 1996;271:13308–13316.
31.
McDonnell DP, Goldman ME. RU486 exerts antiestrogenic
activities through a novel progesterone receptor A form-mediated
mechanism. J Biol Chem. 1994;269:11945–11949.
32. Iafrati MD, Karas RH, Aronovitz M, Kim S, Sullivan TR Jr, Lubahn DB, Donnell TF, Korach KS, Mendelsohn ME. Estrogen inhibits the vascular injury response in estrogen receptor alpha-deficient mice. Nat Med. 1997;3:545–548.[Medline] [Order article via Infotrieve]
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 D. M. Herrington, T. D. Howard, G. A. Hawkins, D. M. Reboussin, J. Xu, S. L. Zheng, K. B. Brosnihan, D. A. Meyers, and E. R. Bleecker Estrogen-Receptor Polymorphisms and Effects of Estrogen Replacement on High-Density Lipoprotein Cholesterol in Women with Coronary Disease N. Engl. J. Med., March 28, 2002; 346(13): 967 - 974. [Abstract] [Full Text] [PDF]
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 D. M. Herrington and K. P. Klein Genome and Hormones: Gender Differences in Physiology: Invited Review: Pharmacogenetics of estrogen replacement therapy J Appl Physiol, December 1, 2001; 91(6): 2776 - 2784. [Abstract] [Full Text] [PDF]
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 R. K. Dubey and E. K. Jackson Estrogen-induced cardiorenal protection: potential cellular, biochemical, and molecular mechanisms Am J Physiol Renal Physiol, March 1, 2001; 280(3): F365 - F388. [Abstract] [Full Text] [PDF]
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 M. POTIER, S. J. ELLIOT, I. TACK, O. LENZ, G. E. STRIKER, L. J. STRIKER, and M. KARL Expression and Regulation of Estrogen Receptors in Mesangial Cells: Influence on Matrix Metalloproteinase-9 J. Am. Soc. Nephrol., February 1, 2001; 12(2): 241 - 251. [Abstract] [Full Text]
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 G. B. Stefano, P. Cadet, C. Breton, Y. Goumon, V. Prevot, J. P. Dessaint, J.-C. Beauvillain, A. S. Roumier, I. Welters, and M. Salzet Estradiol-stimulated nitric oxide release in human granulocytes is dependent on intracellular calcium transients: evidence of a cell surface estrogen receptor Blood, June 15, 2000; 95(12): 3951 - 3958. [Abstract] [Full Text] [PDF]
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Y. K. Hodges, L. Tung, X.-D. Yan, J. D. Graham, K. B. Horwitz, and L. D. Horwitz Estrogen Receptors {alpha} and {beta} : Prevalence of Estrogen Receptor {beta} mRNA in Human Vascular Smooth Muscle and Transcriptional Effects Circulation, April 18, 2000; 101(15): 1792 - 1798. [Abstract] [Full Text] [PDF]
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G. B. Stefano, V. Prevot, J.-C. Beauvillain, P. Cadet, C. Fimiani, I. Welters, G. L. Fricchione, C. Breton, P. Lassalle, M. Salzet, et al. Cell-Surface Estrogen Receptors Mediate Calcium-Dependent Nitric Oxide Release in Human Endothelia Circulation, April 4, 2000; 101(13): 1594 - 1597. [Abstract] [Full Text] [PDF]
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 R. K. Dubey, E. K. Jackson, D. G. Gillespie, L. C. Zacharia, B. Imthurn, and P. J. Keller Clinically Used Estrogens Differentially Inhibit Human Aortic Smooth Muscle Cell Growth and Mitogen-Activated Protein Kinase Activity Arterioscler. Thromb. Vasc. Biol., April 1, 2000; 20(4): 964 - 972. [Abstract] [Full Text] [PDF]
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