Endometriosis: Immunohistochemical analysis of oestrogen and progesterone receptors in endometriotic tissue and endometrium

The objective of the study was to compare the localization andstaining intensity of oestrogen and progesterone receptors inendometrium and endometriotic tissue. Using monoclonal antibodiestowards oestrogen and progesterone receptors, analysis was performedin 63 endometriotic samples from 40 women and compared to endometriumobtained simultaneously from 25 of the women. Using a stainingindex, ‘total immuno-staining score’, calculatedfrom the staining intensity multiplied by the fraction of positivecells, the receptor content was estimated semiquantitatively.The scores for both oestrogen and progesterone receptors werelower in endometriotic epithelial cells than in endometrialepithelial cells, but the differences reached statistical significanceonly for the progesterone receptor. No difference was foundfor stromal cells. There was a significant correlation betweenoestrogen receptor score in endometriotic tissue and in endometrium,but not for progesterone receptor score. In endometrium andvaginal and peritoneal endometriosis, the progesterone receptorscore showed similar values, higher than those in ovarian endometriosis.The data from this large immuno-histochemical study supportprevious results of quantitative steroid receptor analyses,indicating that the regulation of steroid effects, especiallythose of progesterone, differs between endometriotic and endometrialtissue.     

The effect of a levonorgestrel-releasing intrauterine device on human endometrial oestrogen and progesterone receptors after one year of use

Thirty-four women bearing a levonorgestrel-releasing intrauterine device, 20 micrograms/day (LNG-IUD-20), for 12-15 months were recruited. Endometrial biopsies were collected during the late proliferative phase of the cycle (on cycle days 10-12) before (control) and after the use of the IUD for 12 months, and assayed for oestrogen receptors (ER) and progesterone receptors (PR). An immunohistochemical technique with the peroxidase-antiperoxidase detection system (PAP method) was employed. D75 and JZB39 were the primary antibodies for ER and PR respectively. The immunostaining semiquantitative analysis was performed with a computerized microscope image processor, and expressed as 'grey value'. Both endometrial ER and PR populations were significantly lower after insertion of the IUD (P < 0.01) than in control biopsies. The intensity of nuclear staining and the percentage of positively stained cells for ER and PR in women with LNG-IUD were each about 50% of those in control biopsies. The results suggested that LNG released locally from the IUD has a depressive action on the ER and PR, which may contribute to the contraceptive effectiveness of this type of IUD and also to the possible causes of LNG-IUD-induced irregular bleeding and amenorrhoea.     

Physiology: Immunohistochemical distribution of progesterone, androgen and oestrogen receptors in the human ovary during the menstrual cycle: relationship to expression of steroidogenic enzymes

In order to characterize immunohistochemically the possiblein-situ effects of gonadal steroid hormones in the human ovaryduring the menstrual cycle, we immunolocalized progesterone(PR), androgen (AR) and oestrogen (ER) receptors in 50 normalcycling human ovaries, and examined the relationship betweenthese findings and the cellular localization of steroidogenicenzymes including cytochrome P-450 cholesterol side-chain cleavage(P-450scc) enzyme, 3-hydroxysteroid dehydrogenase (3HSD), cytochromeP-450 17-hydroxylase (P-450c17) and cytochrome P-450 aromatase(P-450arom). A large number of stromal cells were positive forAR, regardless of the distance from a follicle. No steroidogenicenzymes were observed in the stromal cells. In the pre-antralfollicle, AR was observed in the theca cells. P-450scc, 3HSDand P-450c17 were sporadically expressed in the theca cellsin relatively large-sized pre-antral follicles. ER was positivein the granulosa cells only in the P-450arom-positive antralor pre-ovulatory follicle, which is likely to be a selectedfollicle. In the corpus luteum, in the period from ovulationto the mid-secretory phase, PR immunoreactivity was observedin a large number of both the luteinized granulosa and the thecacells. All steroidogenic enzymes were observed in all corporalutea, but ER was negative in any corpus luteum. In the atreticfollicle, AR was present in the theca interna cells. P-450scc,3HSD and P-450c17 were observed in the theca interna cells insome atretic follicles. These immunohistochemical findings demonstratedthat steroid hormones locally produced may have important roles,as local regulators, in the human ovary during the menstrualcycle     

Evaluation of oestrogen and progesterone receptor expression in uterine mucosal lymphocytes

Expression of the oestrogen and progesterone receptors on uterinemueosal leukocytes has been examined by dual immunohistology.Neither the oestrogen receptor nor the progesterone receptorwas expressed by lymphocytes, macrophages or the distinctivepopulation of uterine natural killer (NK) cells. Although theaccumulation and survival of these NK cells appears to be hormonallydependent, the effects must therefore be indirect.     

Endometriosis: Oestrogen and progesterone receptors in endometriosis: heterogeneity of different sites

The expression of receptors for the ovarian steroid hormonesoestrogen and progesterone was studied immunohisto-chemicallyusing monoclonal antibodies in samples of endometriosis andendometrium in 22 patients. In nine patients samples of endometriosisfrom more than one site were studied. There was marked heterogeneityin expression of receptors in endometriosis, both when comparinglesions with the corresponding endometrium and also betweensamples of endometriosis collected from different sites withinthe same patient. It wa ssuggested that local environmentalfactors related to the site deotg abd degree if fibrosis ofthe lesions determine the amount of steroid hormone stimylationreaching the lesions and account for the observed differencebetween endometriosis and endometrium and between endometriosislesions of different sites.     

The role of oestrogen and progesterone receptors in gigantomastia

IntroductionGigantomastia is a rare condition characterised by excessive breast growth. The pathophysiology of mammary enlargement varies depending on the type of gigantomastia: gestational, juvenile virginal, or idiopathic. The study aimed at examining the receptor status (oestrogen receptor α (ERα) and progesterone receptor (PR)) of breast tissue in adult women with juvenile or idiopathic gigantomastia.Material and methodsThe study involved 70 women who underwent breast reduction due to juvenile or idiopathic gigantomastia. Control breast specimens were obtained from 18 female cadavers. ERα and PR expressions were detected immunohistochemically in breast gland samples.ResultsCategorised and uncategorised ERα and PR expression did not differ between women with gigantomastia and control women. It was found that in both groups weak (0–30%) ERα and PR expression was the most common. Analysis of categorised data also did not reveal any significant correlations between ERα or PR and the women’s age: for the whole group: p = 0.795 (ERα), p = 0.207 (PR), for women with gigantomastia: p = 0.934 (ERα), p = 0.43 (PR), and for control women: p = 0.638 (ERα), p = 0.805 (PR).ConclusionsGigantomastia is not caused by increased expression of ERα and PR. Analysing abnormal sensitivity of these receptors to hormones may be crucial in establishing the increased risk of breast cancer in women with gigantomastia.     

Uterus and endometrium: Endometrial vascular smooth muscle oestrogen and progesterone receptor distribution in women with and without menorrhagia

This study tested the hypothesis that alterations in the expressionof oestrogen and progesterone receptors (ER and PR) in endometrialvascular smooth musde cells (VSMC) may play a role in the increasedblood loss that occurs during menorrhagla. Subject groups were:controls (n±40), those with menorrhagia (menstrual bloodloss >80 ml, n±39) and patients post-endometrial ablation(n±16). The aims of our study were to describe the changingdistribution of VSMC ER and PR during the menstrual cycle andto look for differences between the three groups. Immunohistochemicaldouble-staining results for VSMC and either ER or PR were highlyvaried, with 0–85% of endometrial arterioles in a biopsysection having alpha smooth muscle actinlER positive cells,and 0–70% demonstrating PR. There were no significantdifferences between controls, menorrhagia or post-ablation specimens(analysis of variance for ER P±0.72; for PR P±0.17).There were also no significant differences between the differentstages of the menstrual cycle when all three groups were combined(analysis of variance for ER P±0.11; for PR P±0.13).The high variability found in this study may mask biologicallyrelevant differences in endometrial vascular ER and PR distributionbetween different groups.     

Endometrial oestrogen and progesterone receptors and their relationship to sonographic appearance of the endometrium

The rapid development of ultrasonographic equipment now permits instantaneous assessment of follicles and endometrium. The sonographic appearance of the endometrium has been discussed in relation to in-vitro fertilization (IVF) cycles. However, a generally agreed view of the relationship of the sonographic appearance to fecundity in IVF cycles has not emerged. We have studied the relationship between steroid receptors and the sonographic appearance of the preovulatory endometrium in natural cycles and ovulation induction cycles. Preovulatory endometrial thickness was not found to be indicative of fecundity, although a preovulatory endometrial thickness of <9 mm related to an elevated miscarriage rate. The preovulatory endometrial echo pattern did not predict fecundity. No relationships were found among endometrial appearance, endometrial steroid receptors and steroid hormone concentrations in serum. Oestrogen or progesterone receptor concentrations were not related to endometrial thickness or to concentrations of serum oestradiol, the only significant correlation being found between the endometrial concentrations of oestrogen and progesterone receptors. The ratio of progesterone:oestrogen receptor concentration was somewhat less in echo pattern B (not triple line) endometrium compared with pattern A (triple line) endometrium. Oestrogen and progesterone receptor concentrations appeared stable on gonadotrophin induction, though fewer numbers were found during clomiphene cycles than in natural cycles. With regard to the distribution of receptor concentration between clomiphene and natural cycles, most women using clomiphene had very low oestrogen receptor populations. Pregnancy rates were low, in spite of high ovulatory rates during clomiphene treatment and were mainly related to low oestrogen receptor concentrations in preovulatory endometrium.     

Endocrinology: Comparative immunohistochemical study of oestrogen and progesterone receptors in the Fallopian tube and uterus at different stages of the menstrual cycle and the menopause

Oestrogen and progesterone are known to require their correspondingsteroid receptors to manifest structural and functional effectsin the Fallopian tube, uterus and other target organs. Thisstudy compares cyclical variations of these receptors in theuterus and in different segments of the Fallopian tube in thesame subjects using an immunocyto-chemical technique. The resultsshow that in the Fallopian tube, isthmic and ampullary epithelialand stromal oestrogen receptors increased in the follicularphase to a peak at mid cycle and then declined in the late lutealphase. The intensity of immunostaining of oestrogen receptorswas less in the Fallopian tube than in endometrial glandularepithelium. The fimbrial end demonstrated an opposite patternof staining to other segments of the tube. Progesterone receptorimmuno-staining was more intense than that for oestrogen receptorsin the follicular phase, and, whereas it disappeared completelyfrom the endometrial glandular epithelium in the late lutealphase, positive staining was clearly visualized in the tubalepithelium and stroma and endometrial stroma at this stage ofthe menstrual cycle. These differences in the steroid receptorcontent may reflect the changing and different functional rolesof these regions and may have important implications on humanreproduction.     

Variant progesterone receptor mRNAs are co-expressed with the wild-type progesterone receptor mRNA in human endometrium during all phases of the menstrual cycle

Progesterone receptor (PR) variant mRNAs in human endometrium could encode proteins with the potential to alter progesterone action in states of normal and abnormal endometrial development. We have assessed the expression levels of mRNA for the wild-type PR and splice variants of PR mRNA lacking exon 4 (del-4 PR), exon 6 (del-6 PR), exons 4 and 6 (del-4&6 PR), and part of exon 4 (del-p4 PR) or part of exon 6 (del-p6 PR) in the human endometrium throughout menstrual cycle development. Eighty-eight endometrial specimens (47 proliferative, 41 secretory) were collected from patients undergoing hysterectomy for benign gynaecologic causes. Measurements by RT-PCR indicated that mRNAs for wild-type PR, and splice variants del-4 PR, del-6 PR, del-4&6 PR, del-p6 PR, and a novel del-p4 PR were detected in all endometrial specimens throughout the menstrual cycle. Higher levels of wild-type PR and all PR variant mRNAs were found in the early and mid-proliferative endometrial phases than in secretory endometrium. The relative expression of mRNA for all PR variants compared to wild-type PR mRNA, however, did not change through all stages of endometrial development. We, therefore, found no evidence of differential co-expression of the PR variants compared with wild-type PR during normal menstrual development. Future studies will determine if the expression profile of PR variant mRNAs will be different in the endometrium of patients with infertility, recurrent pregnancy loss, or endometrial adenocarcinoma.     

Early changes of affinity and binding sites of progesterone and oestrogen receptors in decidua exposed to RU 486.

Sixty patients with 6-7 weeks of amenorrhoea were randomly allocated to three groups. The women in the first group (control) took a placebo 24 h before undergoing a vacuum aspiration. The patients in the second and third groups were given 200 mg of RU 486 orally, 12 or 24 h before surgical interruption of their pregnancy. Decidua were collected and frozen in liquid nitrogen. By Scatchard plot analysis, the number of cytosolic binding sites (1798 +/- 803 fmol/mg DNA) of progesterone in decidua in the control group was significantly reduced (P less than 0.01) to 696 +/- 408 or 626 +/- 179 fmol/mg DNA by RU 486 treatment for 12 or 24 h respectively. The dissociation constants of both cytosolic and nuclear progesterone receptors in RU 486-exposed decidua were increased (P less than 0.01). The number of nuclear binding sites of oestrogen receptor was significantly higher (P less than 0.05) in decidua with RU 486 treatment for 12 h (178 +/- 77 fmol/mg DNA) compared to the control (89 +/- 32 fmol/mg DNA). The results suggest that RU 486 might regulate progesterone and oestrogen receptors in the decidua of early human pregnancy, either directly or indirectly.     

Reversible suppression of menstruation with progesterone antagonists in rhesus macaques

BACKGROUND: A reliable means of menstrual suppression would greatly improve the quality of life for women. Information is lacking on the direct endometrial effects and appropriate dosages of new antiprogestins that may be useful for this purpose. METHODS: The current work evaluated three different systems in macaque monkeys. First, the range of doses of two relatively new antiprogestins, ZK 137 316 and ZK 230 211, that would block progesterone action directly on the endometrium in artificially cycled, spayed rhesus macaques; second, the direct endometrial effects of ZK 230 211, a type III antiprogestin; and third, investigation of whether endometrial-suppressive doses administered chronically to intact, cycling monkeys could be used for reversible, menstrual suppression. RESULTS: The results in naturally cycling animals showed that ZK 137 316 blocked menstruation in all animals, but doses of 0.05 mg/kg blocked ovulation in 55.5% of animals and doses of 0.1 mg/kg blocked ovulation in 66.6% of the animals. However, all doses of ZK 230 211 that blocked menstruation also blocked ovulation. All progesterone antagonist (PA)-treated animals, regardless of dose, maintained normal follicular phase concentrations of oestradiol and returned to normal menstrual cyclicity within 15--41 days post-treatment.Therefore ZK 137 316, depending on dose, can allow ovulation but block menstruation, while ZK 230 211, a much more potent PA, blocks both ovulation and menstruation at all effective doses. Both PAs block unopposed oestrogenic action on the endometrium through their antiproliferative effects. CONCLUSIONS: Reversible amenorrhoea can be achieved with these two PAs, and they can protect the endometrium from the effects of unopposed oestrogen whether or not ovulation is blocked. Chronic, low dose PA treatment may provide a new option for women who wish to suppress their menstrual periods.     

Immunohistochemical localization of progesterone receptor in human decidua of early pregnancy.

Rat monoclonal antibodies to human progesterone receptor were used for immunolocalization studies in human decidua of early pregnancy. Frozen sections of 42 specimens of decidua were stained by the peroxidase-antiperoxidase method (PAP). Progesterone receptor was localized exclusively in the nuclei of decidual and myometrial cells with no specific staining in the cytoplasm. In the decidualized endometrium, stroma were always positively stained. Smooth muscle, pericyte and endothelial cells of blood vessels were extensively stained. Glandular epithelia showed variation in staining, which was positive in the basal but very weak or negative in the superficial layer of the decidua. No specific staining could be detected in the control sections. Of special interest was the positive staining of the endothelium of decidual blood vessels, a finding which has not been reported previously. The cells of the inner lining of vessels that stained with the antiprogesterone receptor antibodies were also Factor VIII positive, thus confirming the endothelial nature of these cells. It is concluded from these results that endothelial cells from human first trimester decidua express progesterone receptors.     

Immunolocalization of oestrogen and progesterone receptors in prostatic hyperplasia and carcinoma

Oestrogen receptors and progesterone receptors were immunolocalized in 19 patients with benign prostatic hyperplasia and in 26 patients with prostatic carcinoma. Immunohistochemistry was performed on tissue that had been fixed in 8% paraformaldehyde and then paraffin-embedded, using microwave irradiation for antigen retrieval. Oestrogen receptor expression was observed exclusively in the stromal cells of six out of 26 (23%) patients with prostatic carcinoma, but in none of the cells of patients with benign prostatic hyperplasia. Progesterone receptor expression was detected in 16 of 19 (84%) and 17 of 19 (89%) of the epithelial cells and stromal cells of patients with benign prostatic hyperplasia, respectively. In patients with prostatic carcinoma, progesterone receptor immunoreactivity was observed in 12 of 20 (46%) and 20 of 26 (77%) of the carcinoma and stromal cells of prostatic carcinoma, respectively. The ratio of epithelial cells with progesterone receptor immunoreactivity corresponded well with that of stromal cells with immunoreactivity in patients with benign prostatic hyperplasia. However, the ratio of stromal cells with progesterone receptor immunoreactivity was much higher than that in carcinoma cells in patients with prostatic carcinoma. Immunolocalization patterns or the ratio of the cells with progesterone receptor immunoreactivity did not significantly correlate with histological differentiation or patient's age in carcinoma cases. However, patients with advanced surgical stages of disease demonstrated a significantly smaller number of carcinoma and stromal cells with progesterone immunoreactivity in patients with prostatic carcinoma. These results suggest that oestrogens do not have a direct effect on the biological behaviour of benign prostatic hyperplasia and prostatic carcinoma, but that progesterone appears to play a role in the pathogenesis of benign prostatic hyperplasia and prostatic carcinoma.     

Pregnancy: The effect of RU486 on progesterone and oestrogen receptor concentration in human decidua on early pregnancy

Concentrations of progesterone receptor (PR) and oestrogen receptor(ER) were measured by radioligand assay in decidual tissue ofwomen undergoing termination of early pregnancy (amenorrhoeaup to 49 days). Pregnancies were terminated by vacuum aspirationat 12 or 36 h after oral administration of placebo or antiprogestinRU486 in different doses. Treatment with RU486 decreased decidualPR content, the effect being observed at 12 h as well as at36 h after 600 mg RU486 and at 36 h after 3 x 25 mg RU486 givenat 12 h intervals. PR concentration 12 h after a single doseof 25 mg RU486 was not affected. ER content was unchanged at12 h after RU486 but increased 36 h after 600 mg and 3 x 25mg RU486. Our data suggest that apart from blocking progesteroneaction, RU486 may exert its abortifacient effect through decreasingthe PR concentration. The simultaneous decrease of PR concentrationand an increase of ER concentration changes the balance betweenthem in favour of ER, which might also play a role in the abortifacienteffect of RU486.     

Oestrogen and progesterone do not regulate the expression of retinoic acid receptors and retinoid 'X' receptors in human endometrial stromal cells in vitro

Elucidation of the gene structure for retinoic acid receptor-(RAR-) has suggested a potential role for oestrogen in regulatingthe expression of RAR-. We have previously shown that all threeRAR types are expressed in human endometrial stromal cells invitro and that RAR- expression is induced in response to retinoicacid. The aim of this study was to ask whether oestradiol andprogesterone could play a part in regulating the expressionof RARs in human endometrial stromal cells and to establishthe patterns of expression of a related group of nuclear retinoidreceptors, retinoid ‘X’ receptors (RXRs) and theirpotential for regulation by steroid hormones. The RAR expressionpatterns of endometrial stromal cells, grown in steroid-freemedium, did not change in response to the presence of steroidhormones. Furthermore, the retinoic acid-mediated inductionof RAR- was not affected by oestradiol or progesterone, andwas dependent on the continued presence of retinoic acid. Ofthe three RXR types, only RXR- was detectably expressed in stromalcells in vitro and the expression of RXR- did not change inresponse to steroid hormones or retinoic acid. These data indicatethat oestradiol and progesterone are not important in the regulationof RAR and RXR expression in human endometrial stromal cells.     

Demonstration of oestrogen and progesterone receptors in freeze-dried, paraffin-embedded sections of breast cancer

Immunohistochemical evaluation of oestrogen and progesterone receptors is of importance in evaluating human breast tumours. Staining techniques can be performed on snap-frozen, cryostat-cut tissues or, as recently reported, on formalin-fixed, paraffin-embedded tissues. These methods are, however, limited by several drawbacks, including difficulties in retrospective studies and in storage of the material, and the relatively high frequency of false negative results for chemically fixed specimens. We therefore investigated the application of freeze-drying technology to assess the feasibility and reliability of this technique as an alternative method for diagnostic breast pathology. Morphological and immunohistochemical studies were performed on snap-frozen, freeze-dried and paraffin-embedded tissue obtained from 16 cases of benign and malignant breast neoplasms. Our results showed good preservation of tissue morphology, similar to standard formalin fixation, and excellent preservation of antigenic reactivity of nuclear receptors, comparable to that obtained with cryostat sections. We therefore suggest that freeze drying and paraffin embedding of frozen tissue blocks is equivalent or even preferable to formalin fixation for the demonstration of oestrogen and progesterone receptors, at least in the case of small tumours.     

The effect of RU 486 on progesterone receptor in villous and extravillous trophoblast

A total of 60 women with 6–7 weeks' amenorrhoea were randomlyallocated to three groups. The women in the first group (control)took a placebo 24 h before undergoing vacuum aspiration. Thesubjects in the second and third groups were given orally 200mg of RU 486, 12 or 24 h before surgical interruption of theirpregnancy. Villi and decidua were collected and frozen in liquidnitrogen. There were no significant differences in villous cytosolicsteroid concentrations between the control and RU 486-treatedgroups. The RU 486 concentration was lower in villous cytosolthan in decidua and serum. The immunostaining of progesteronereceptor in villous and extravillous trophoblast was weak andlocalized in both cytoplasm and nucleus. RU 486 treatment hadno effect on the immunostaining of progesterone receptors introphoblast populations except in the placental bed giant cells,where the weak and diffuse progesterone receptor staining inthe cytoplasm of controls seemed to be concentrated in the nucleusafter RU 486 treatment. In conclusion, RU 486 might have noeffect on the immunostaining of progesterone receptor and onsteroid concentrations in villi in vivo.     

Immunolocalization of oestrogen and progesterone receptors in the human decidua in relation to prolactin production

The distribution of oestrogen and progesterone receptors withinthe decidualized stroma of the uterus was examined in earlyand term human pregnancy and the results related to the effectof oestradiol and progesterone on prolactin production by deciduain vitro. In early pregnancy progesterone receptors were presentin the nucleus of decidualized cells of both the capsularisand parietalis but not in glandular cells. In contrast at termprogesterone receptors were located within the cytoplasm ofdecidual cells. Oestrogen receptors were detected only in thenucleus and were present in greater amounts in decidua capsularisthan parietalis in early pregnancy, but were not detectablein term decidua. Both oestrogen and progesterone receptors werepresent in the nuclei of cells of arterioles within the decidua.In early pregnancy prolactin production decreased during in-vitroculture of decidua parietalis but was maintained in deciduacapsularis, associated with an increase in progesterone productionby the decidua capsularis. In term decidua, prolactin productionin vitro was only stimulated by a combination of oestradioland progesterone. These results suggest, firstly, that maintaineddecidualization and prolactin production by decidua capsularisduring treatment of women in early pregnancy with the anti-progestinmifepristone is not due to an absence of progesterone receptor;secondly, there is a shift in immunoreactive progesterone receptorin decidual cells from the nucleus in early pregnancy to thecytoplasm in term pregnancy. This may indicate an alterationin the action of progesterone around the time of parturition;and thirdly, in term decidua, progesterone, apparently actingthrough the cytoplasmic receptor, is active in increasing prolactinproduction in vitro only when combined with oestradiol.