Detection of estrogen receptor mRNA in human uterus

Parl, Fritz F.; Schonbaum, Christopher P.; Cox, Diana L.; Cavener, Douglas R.

doi:10.1016/0303-7207(87)90049-9

Cited by 11 publications

(5 citation statements)

References 25 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…These findings on the location of ER mRNA in the myometrium and endometrium are consistent with previous localization of ER protein using immunocytochemistry [25][26][27] as well as localization of ER mRNA in both human [28] and rhesus monkey [29] uterine tissues. The use of a human ER cDNA probe for the in situ hybridization studies was validated by Northern blot analysis, which showed that the probe hybridized to a single RNA at the expected mass.…”

Section: Discussionsupporting

confidence: 91%

Regulation of the Estrogen Receptor and its Messenger Ribonucleic Acid in the Ovariectomized Sheep Myometrium and Endometrium: The Role of Estradiol and Progesterone1

Wen

Owiny

et al. 1996

Biology of Reproduction

View full text Add to dashboard Cite

Estrogen receptor (ER) mRNA is dramatically increased in sheep myometrium and endometrium during glucocorticoid-induced premature labor and term spontaneous labor. However, the underlying mechanism for the up-regulation of uterine ER in labor is still unknown. We used ovariectomized (OVX) non-pregnant sheep to analyze the role of estradiol and progesterone in the regulation of myometrial and endometrial ER protein and ER mRNA in vivo. Twenty-one OVX ewes were treated with saline (n = 6), or with estradiol infused i.v. for 2 days (50 micrograms/day, n = 5), or with an intravaginal progesterone sponge for 10 days (containing 0.3 g progesterone, n = 5), or with an intravaginal progesterone sponge for 10 days with estradiol (50 micrograms/day) administered on Days 9 and 10 with the progesterone sponge still in place (n = 5). The ER protein concentration in both cytosolic and nuclear compartments, analyzed by Western blot, increased significantly (P < 0.05) in the myometrium after estradiol treatment, while progesterone alone had no detectable effect on ER level. Elevated ER protein was observed only in the nuclear fraction of endometrium. However, when estradiol was given together with progesterone treatment, progesterone antagonized the up-regulatory effect of estradiol on the ER level both at the endometrium and myometrium. The changes in cellular ER mRNA followed the pattern observed at the ER protein level. Estrogen receptor mRNA was elevated significantly (p < 0.01) only in estradiol-treated ewes. Expression of the ER gene in ewes receiving progesterone alone or progesterone combined with estradiol was similar to that of the control group. From these observations we conclude that ER gene expression and active ER synthesis in nonpregnant sheep myometrium and endometrium are estradiol-dependent. Progesterone antagonizes this estrogen action. Progesterone down-regulated the elevated ER mRNA when used together with estradiol. In situ hybridization showed that ER mRNA was evenly distributed in the smooth muscle cells and blood vessels of the myometrium and the epithelial cells of the glands in endometrium. In conclusion, we have observed estradiol-dependent activation of ER gene expression as well as active ER synthesis in the nonpregnant sheep myometrium and endometrium. Progesterone acted as an antagonist of estradiol on ER gene expression.

show abstract

Section: Discussionsupporting

confidence: 91%

Regulation of the Estrogen Receptor and its Messenger Ribonucleic Acid in the Ovariectomized Sheep Myometrium and Endometrium: The Role of Estradiol and Progesterone1

Wen

Owiny

et al. 1996

Biology of Reproduction

View full text Add to dashboard Cite

show abstract

“…We detected hybridisation to a single oestrogen receptor RNA band of approximately 6.2kb in extracts of RNA prepared from MCF-7 breast cancer cells, all the positive breast tumours and a uterus from a premenopausal woman. A 6.2 kb RNA was detected previously in MCF-7 cell RNA (Walter et al, 1985) whereas others detected a 4.2kb oestrogen receptor RNA in the uterus from a premenopausal woman (Parl et al, 1987). We did not detect the 3.7kb oestrogen receptor RNA found by Barrett-Lee et al (1987) in both MCF-7 cell and breast tumour RNA.…”

Section: Discussioncontrasting

confidence: 41%

“…The recent isolation and cloning of the oestrogen receptor cDNA by Walter et al (1985) has now made it possible to study oestrogen receptor mRNA expression. Parl et al (1987) logically. RNA was also prepared from combined endometrium and myometrium from the uterus of a premenopausal woman.…”

mentioning

confidence: 99%

Measurement of oestrogen receptor mRNA levels in human breast tumours

Henry

Nicholson²,

Farndon³

et al. 1988

Br J Cancer

View full text Add to dashboard Cite

Summary A sensitive single-stranded hybridisation probe for the oestrogen receptor mRNA was synthesised using T3 polymerase from oestrogen receptor cDNA cloned in the Bluescript vector. This probe was used to measure oestrogen receptor mRNA in total RNA extracted from breast tumours. Oestrogen receptor mRNA was detected in 41 of 47 (87%) tumours whereas cytosolic oestrogen receptor protein was detected in only 18 out of 39 (46%). There was a significant correlation between the levels of the oestrogen receptor, as measured by 3H-oestradiol binding, and the oestrogen receptor mRNA. Breast cancer has long been known to be oestrogen responsive and the expression of the oestrogen receptor in a proportion of tumours is well recognised (McGuire et al., 1975). Different studies find the proportion of breast tumours expressing oestrogen receptor to be variable, ranging from 60% to 80% (Hawkins, 1985). Most studies indicate that patients with oestrogen receptor-positive tumours have better disease-free, post-relapse and overall survival, compared to those with oestrogen-receptor negative tumours (reviewed by Hawkins, 1985). A higher proportion of oestrogen receptor-positive tumours respond to antioestrogen therapy: paradoxically, a small number of oestrogen receptor-negative tumours also respond (Desombre et al., 1978).A variety of technique' have been used to determine oestrogen receptor protein levels in breast tumours, including tritiated hexoestrol binding in vivo (Folca, 1961), sedimentation assays (Desombre et al., 1978), dextran-coated charcoal assays (Korenman & Dukes, 1970), radio-immunoassay (Thorpe, 1987 logically. RNA was also prepared from combined endometrium and myometrium from the uterus of a premenopausal woman. Patient age ranged from 24 to 95 years: 14 were less than 50 years old and considered to be premenopausal. RNA extractionRNA was extracted from the 47 tumour biopsies and the uterus using a modification of the method of Auffrey & Rougeon (1980). Tumours frozen in liquid nitrogen were reduced to a powder using a Braun mikro dismembrator. Tumour powder was transferred to a Corex centrifuge tube, suspended in 5ml of 3M LiCl, 6M urea, 0.5% SDS and 50mM sodium acetate (pH5.6) and the DNA was sheared with an Ultraturrax homogeniser. RNA was precipitated at 4°C for 18h and recovered by centrifugation at 10,OOOg for 20 min. The supernatant was discarded and the resultant pellet was washed in 3 ml of 3 M LiCl, 6 M urea and 50mM sodium acetate and recentrifuged at 10,000 g for 0 min. The pellet was then resuspended in 1.125ml lOmM Tris buffer pH 8.0, 0.2% SDS, at room temperature, Tris buffer pH 8.0 and EDTA were added to 0.1 M and 5mM, respectively in 1.25ml total volume and then extracted with phenol:chloroform (50:50) and chloroform:isoamyl alcohol (24:1). The organic phases were back extracted with 1.25 ml 100mM Tris pH9. Following the addition of 6.25ml 100% ethanol and 50 pl 5 M NaCl, RNA was precipitated at -20°C. After centrifugation, the pellet was washed in 70% cold ethanol, recentrifuged, dried...

show abstract

“…The use of a human ER cDNA probe and a human ER antibody for this study was validated by Northern and Western analyses, which showed that the probe and antibody hybridized to a single RNA and specific protein species at the expected masses. There have been few attempts to use in situ hybridization to examine ER mRNA content and distribution in histologic sections of the nonpregnant uterus, and none in the pregnant sheep uterus [29,30]. We observed that ER and ER mRNA were present mainly in the smooth muscle cells and the cells around the blood vessels in the myometrium, indicating an involvement of ER both in myometrial contractility and in regulation of uterine blood flow during pregnancy.…”

Section: Discussionmentioning

confidence: 80%

Effects of glucocorticoids on estrogen receptor messenger ribonucleic acid in the pregnant ovine myometrium in vivo and in vitro

Wen

Derks

Nathanielsz

1996

Biology of Reproduction

View full text Add to dashboard Cite

We recently reported that estrogen receptor (ER) mRNA is dramatically increased in the sheep myometrium during cortisol-induced premature labor and term spontaneous labor. In this study, we compared myometrial ER mRNA and ER protein levels in tissues from pregnant sheep during infusions of two different glucocorticoids (dexamethasone and betamethasone). Tissues from glucocorticoid-infused sheep not yet in labor were compared with tissues from sheep in labor. The population of ER mRNA-containing cells in myometrium obtained from sheep in labor was determined by in situ hybridization and compared with that of sheep not in labor in order to understand how the cellular distribution of ER mRNA changed during labor. Finally, in vitro myometrial cell culture and immunolocalization of glucocorticoid receptor (GR) were used to determine 1) the effects of different steroids on ER mRNA content, 2) whether or not the myometrium is a functional site for glucocorticoid action, and 3) whether the change in ER mRNA content in the myometrium might be induced by glucocorticoid acting through GR. Increased ER mRNA and protein were found only in the myometrium associated with labor. In situ hybridization of ER mRNA and immunolocalization of ER protein showed that ER mRNA and protein were mainly located in the smooth muscle cells and endothelial cells of blood vessels. In vitro treatment of cultured myometrial cells with hydrocortisone resulted in a 3.5-fold increment in ER mRNA. In contrast, there was no obvious change in ER mRNA when myometrial cells were treated in vitro with either estradiol (10 nM) or progesterone (100 nM) for 24 h. GR was localized exclusively in the pregnant sheep myometrium. We conclude that 1) increased ER protein and ER mRNA in sheep myometrium are strongly associated with labor; 2) an increase in the population of ER-positive cells is associated with the increment of ER mRNA during glucocorticoid-induced labor; 3) in the pregnant sheep myometrium, both the smooth muscle cells and endothelial cells of blood vessels are positive for ER protein ER mRNA; and 4) hydrocortisone is a potent stimulus in vitro for increasing ER mRNA content, presumably acting through the GR present in the pregnant sheep myometrium.

show abstract

Detection of estrogen receptor mRNA in human uterus

Cited by 11 publications

References 25 publications

Regulation of the Estrogen Receptor and its Messenger Ribonucleic Acid in the Ovariectomized Sheep Myometrium and Endometrium: The Role of Estradiol and Progesterone1

Regulation of the Estrogen Receptor and its Messenger Ribonucleic Acid in the Ovariectomized Sheep Myometrium and Endometrium: The Role of Estradiol and Progesterone1

Measurement of oestrogen receptor mRNA levels in human breast tumours

Effects of glucocorticoids on estrogen receptor messenger ribonucleic acid in the pregnant ovine myometrium in vivo and in vitro

Contact Info

Product

Resources

About