S-phase fractions of breast carcinomas. Relationships to morphology, estrogen and progesterone receptors, and early recurrence

Analysis of the S-phase fractions (SPF) measured by in vitro thymidine labeling, morphological appearances, and estrogen receptor (ER) assays of primary invasive breast carcinomas demonstrated several interrelationships. Lobular, mucinous, tubular, and adenocystic carcinomas consistently had low SPF and were usually positive for ER. The same was true for the carcinomas of no special histologic type [the not otherwise specified (NOS) group of E. R. Fisher including "infiltrating ductal" and undifferentiated carcinomas] with minimal anaplasia. Medullary, atypical medullary, and morphologically unclassifiable carcinomas with marked nuclear anaplasia nearly always had high SPF and were usually negative for ER. High SPF was associated with advanced stages of carcinoma initially or with early recurrence following mastectomy.     

Hepatobiliary and pancreatic mucinous cystadenocarcinomas with mesenchymal stroma: analysis of estrogen receptors/progesterone receptors and expression of tumor-associated antigens

In an attempt to establish a standardized rating system for CT of the paranasal sinuses, the Committee on Rhinology and Paranasal Sinus Disease of the American Academy of Otolaryngology-Head and Neck Surgery instituted a protocol for the review of sinus CT scans at six international sites. Fifty identical scans were rated by four otolaryngologists at each site according to five established sinus CT staging systems. Twenty of 24 reviewers repeated the rating session at least 1 week later to determine intrarater variability. The number of CT scans that could not be classified by a particular rating system ranged from 1.3% to 5.5%. The range of intrarater agreement (kappa = 0.39 to 0.74) exceeded that of interrater agreement (kappa = 0.18 to 0.49). A skewed distribution of CT scans resulted in a system with high rater agreement but poor ability to differentiate among disease states. The use of a numeric rating system to assign a score to each scan produced a comprehensive and disease-sensitive system, but one with low rater agreement. A precise definition of mucosal thickening in terms of millimeters appeared to enhance the raters' ability to assign stage and improve a system's comprehensiveness and reproducibility. On the basis of these findings, recommendations are made for the use of CT rating systems to study clinical outcomes in patients with chronic sinusitis.     

Immunohistochemical analysis of distribution of estrogen receptors and progesterone receptors in the postmenopausal endometrium

OBJECTIVE: To examine the expression of sex steroid receptors (ER: estrogen receptor; PR: progesterone receptor) in the postmenopausal endometrium (PMEM) and the relationship to clinical data for studying its characters. METHODS: The immunohistochemical reactivity of the PMEM was studied using monoclonal antibodies against ER and PR, in 33 postmenopausal patients. RESULTS: The endometrium was thicker in patients who were postmenopausal for 1 to 10 years (1.48 +/- 1.31 mm) than in patients who were postmenopausal for more than 10 years (0.79 +/- 0.37 mm)(p < 0.05). Among the 33 postmenopausal endometrial samples, ER positivity was found in the glands in 26 cases (78.8%) and PR positivity was detected in 18 cases (54.5%). The average age of the patients with ER positive reactivity in the glands (61.69 +/- 7.26 years) was significantly lower than that of the patients with ER negative reactivity (66.00 +/- 3.56 years)(p < 0.05). Furthermore, the endometrial thickness of the patients with ER or PR positive reactivity in the glands (1.24 +/- 1.09 mm and 1.47 +/- 1.20 mm, respectively) was significantly greater than that of the patients with ER or PR negative reactivity (0.67 +/- 0.26 mm and 0.70 +/- 0.40 mm, respectively)(p < 0.05). CONCLUSION: ER in the glands of the PMEM was determined to decrease gradually with increased aging. The presence of ER and PR in the gland cells seemed likely to determine the thickness of the PMEM.     

Primary mucinous carcinoma of the skin. Report of case

The authors report a case of a primary mucinous carcinoma of the skin. This is a rare adnexial tumor arising from the eccrine secretory coils. The histologic characteristic finding of this lesion is the heap of abundant mucinous material (sialomucin) in the dermis in which rare epithelial islands are free-floating. In this study we also value the importance of a correct diagnosis for an appropriate therapy.     

Study on estrogen and progesterone receptors in endometriosis and adenomyosis

Estrogen and progesterone receptors (ER and PR) in 18 cases of ovarian endometriosis and 13 cases of adenomyosis were determined with dextran coated charcoal (DCC) method. The levels of ER and PR in those specimens were lower than those of normal endometrium. Among the 18 cases of ovarian endometriosis, 6 (33.4%) were negative for PR, which accounted for the unsatisfactory results of progesterone treatment for some endometriosis. In the 13 cases of adenomyosis there were 10 (76.9%) showing positive PR. It is suggested that the hormone therapy may be useful to treat those young patients with adenomyosis instead of surgery. The correlation of the ER and PR levels, the treatment and prognosis in endometriosis and adenomyosis are worth further studying.     

Evidence that estrogen-receptor-negative, progesterone-receptor-positive breast and ovarian carcinomas contain estrogen receptor

A small number of primary metastatic breast carcinomas are estrogen-receptor-negative and progesterone-receptor-positive (ER-, PGR+) under the normal ligand-binding assay or sucrose density gradient conditions. Among more than 500 tumors analyzed in this laboratory over a year and a half, 28 cases fit this category, 18 of which were patients 51 years of age or younger (Group A) and 7 were patients over the age of 56 (Group B), The ages of three patients were unknown (Group C). By treatment of each of those tumor cytosols with dextran-coated charcoal before the assay was done, 13 of group A became positive (ER range 10-87 fmol/mg protein); 1 was borderline (ER 3-9 fmol/mg protein); 1 became positive only on sucrose gradient determination, and 2 remained negative. In comparison, two patients in group B shifted from borderline ER to ER+ and only one ER- became ER+ at 10 fmol/mg protein. The data provide additional rationale for determining both ER and PGR in all patients, and have obvious implications for the need of standard methods of determining ER and PGR in the prognosis of women with breast cancer.     

Methylation of estrogen and progesterone receptor gene 5' CpG islands correlates with lack of estrogen and progesterone receptor gene expression in breast tumors

Hormonal factors have a profound influence on the development, treatment, and outcome of breast cancer. The absence of steroid hormone receptors is highly correlated with resistance to antihormonal treatments. Work in cultured human breast cancer cell lines has shown that the absence of estrogen receptor (ER) gene expression in ER- cells is associated with extensive methylation of the ER gene 5' CpG island, and treatment with agents that demethylate the ER gene CpG island results in the production of functional ER protein. The current study shows that CpG islands in the 5' region of the ER and progesterone receptor (PR) genes are methylated in a significant fraction of primary human breast cancer tissues. The ER CpG island is methylated at the methylation-sensitive NotI restriction site in 9 of 39 (25%) of primary ER- breast cancers but remains unmethylated in 53 ER+ breast cancers and 9 normal breast specimens. Three methylation-sensitive restriction sites in the PR gene CpG island are not methylated in normal breast specimens and PR+ human breast cancers but are hypermethylated in 40% of PR- human breast tumors. These data demonstrate that methylation of the ER and PR gene CpG islands is associated with the lack of ER and PR gene expression in a significant fraction of human breast cancers.     

Mutual and intercompartmental regulation of estrogen receptor and progesterone receptor expression in the mouse uterus

The epithelial and stromal compartments of the uterus undergo significant estrogen- and progesterone (P4)-induced changes during the estrous cycle. While in the adult mouse, epithelial proliferation and stromal inflammation are induced by estrogen, P4 is antiproliferative in the epithelium and both proliferative and anti-inflammatory in the stroma. In light of these compartmentally varying roles, we have immunohistochemically examined estrogen and P4 regulation of the expression of their receptors (ER and PR) and their epithelial target gene lactoferrin (LF) in wild-type and PR null mutant mice. We demonstrate that estrogen exerts compartment-specific effects on the expression of ER, resulting in decreased levels of stromal and glandular epithelial (GE) ER and increased luminal epithelial (LE) and myometrial ER. Estrogen also has dual effects on PR expression, decreasing levels in the LE while at the same time increasing levels in the stroma and myometrium. Estrogen and P4 together mediate their effects in part through the ability of P4 to selectively inhibit myometrial ER expression while preserving GE expression. We also demonstrate a general negative feedback by P4 on PR expression that is most prominent in the GE. Finally, we demonstrate using the estrogen- and P4-responsive epithelial target gene LF that the differential regulation of PR in the glandular and luminal epithelium results in different functional responses of these compartments to P4. Together, our data indicate that the pleiotropic effects of estrogen and P4 in the adult mouse uterus are mediated by complex hormonal interregulation of ER and PR in specific uterine compartments.     

Differential spatiotemporal regulation of lactoferrin and progesterone receptor genes in the mouse uterus by primary estrogen, catechol estrogen, and xenoestrogen

Many xenobiotics are considered reproductive toxins because of their ability to interact with the nuclear estrogen receptors (ERalpha and ERbeta). However, there is evidence that these xenobiotics can regulate gene expression in the reproductive targets by mechanisms that do not involve these ERs. To examine this further, we compared the effects of estrogenic (o,p'-DDT [1-(o-chlorophenyl)-1-(p-chlorophenyl)2,2,2-trichloroethane] and Kepone, chlordecone) and nonestrogenic (p,p'-DDD [1,1-dichloro-2,2-bis(p-chlorophenyl)ethane], a metabolite of p,p'-DDT) xenobiotics with those of 17beta-estradiol (E2) and 4-hydroxyestradiol-17beta (4-OH-E2), a catechol metabolite of E2, on uterine expression of lactoferrin (LF) and progesterone receptor (PR). These genes are estrogen responsive in the mouse uterus. Normally, LF is expressed in the uterine epithelium, whereas PR is expressed in both the epithelium and stroma in response to estrogenic stimulation. Ovariectomized mice were injected with xenobiotics (7.5 mg/kg), E2 (10 microg/kg), 4-OH-E2 (10 microg/kg), or the vehicle (oil, 0.1 ml/mouse), and uterine tissues were processed for Northern blot and in situ hybridization. The pure antiestrogen ICI-182780 (ICI; 1 or 20 mg/kg) was used to interfere with estrogenic responses that were associated with the ERs. The results of Northern and in situ hybridization demonstrated increased uterine levels of PR and LF messenger RNAs (mRNAs) by all of these xenobiotics, but quantitatively the responses were much lower than those induced by E2 or 4-OH-E2. The results further showed that the E2-inducible epithelial LF mRNA accumulation was markedly abrogated by pretreatment with ICI (20 mg/kg). In contrast, this treatment retained the epithelial expression of PR mRNA, but down-regulated the stromal expression. In contrast, ICI had negligible effects on LF and PR mRNA responses to 4-OH-E2, indicating that this catechol estrogen exerted its effects primarily via a mechanism(s) other than the ERs. The heightened accumulation of LF mRNA in the epithelium in response to Kepone and o,p'-DDT was also severely compromised by pretreatment with ICI, but this antiestrogen had little effect on responses to p,p'-DDD. Similar to E2, Kepone increased the expression of PR mRNA in both uterine epithelium and stroma. However, pretreatment with ICI decreased stromal cell expression, whereas epithelial cell expression remained unaltered or increased. These responses were not noted in mice treated with o,p'-DDT or p,p'-DDD. Collectively, the results demonstrate that catechol estrogens or xenobiotics can alter uterine expression of estrogen-responsive genes by mechanisms that are not totally mediated by the classical nuclear ERs, and these alterations are cell type specific. We conclude that an interaction of a compound with the nuclear ERalpha and/or ERbeta is not an absolute requirement for producing specific estrogen-like effects in the reproductive target tissues.     

Immunohistochemical detection of estrogen and progesterone receptors in spermatozoa of infertile men

OBJECTIVE: To evaluate the clinical implication of sex steroidal actions on human spermatozoa. PATIENTS AND METHODS: Human spermatozoa were obtained from 20 males. Immunohistochemical staining for estrogen receptors (ER) and progesterone receptors (PR) was conducted by the avidin-biotin-peroxidase complex method. RESULTS: Positive immunohistochemical staining of ER and PR was observed in all 8 cases of fertile couples. From infertile couples, ER were detected in 8/12 cases, including 3 with normozoospermia, 2 with oligozoospermia, 1 with asthenozoospermia, and 2 with teratozoospermia, but not in 4/12 cases, 1 with normozoospermia, 1 with asthenozoospermia, and 2 with oligoasthenozoospermia. PR were undetectable in all cases of negative ER and in 1 case (normozoospermial) of positive ER. CONCLUSION: Negative ER and/or PR in spermatozoa from infertile couples might be involved in cases of male infertility.     

Coronary artery and cultured aortic smooth muscle cells express mRNA for both the classical estrogen receptor and the newly described estrogen receptor beta

Estrogens exhibit potent anti-atherogenic effects through mechanisms which may involve direct effects on the artery. The existence of the classical estrogen receptor (ERalpha) in vascular tissues has been established. Recently a new estrogen receptor (ERbeta) has been discovered which represents a distinct gene product with homology to the classical ERalpha. The purpose of the present study was to determine if ERbeta mRNA is expressed in vascular tissues of female and male primates. Oligonucleotide primers were developed for the specific RT-PCR amplification of ERalpha or ERbeta mRNA. RT-PCR products of the appropriate size for ERalpha and for ERbeta were observed after amplification of RNA isolated from coronary arteries of both male and female cynomolgus monkeys. Similar results were obtained from cultured aortic smooth muscle cells and from monkey reproductive tissues such as ovary and uterus. The relative expression of ERbeta to ERalpha mRNA was greatest in ovary, on the same order of magnitude in monkey vascular tissues and uterus, while the human breast cancer cell line MCF-7 exhibited a very low level of ERbeta relative to ERalpha. Sequence analysis of isolated RT-PCR products showed >95% similarity between the monkey and the published human sequences for both ERalpha and ERbeta. These findings suggest that estrogen may influence vascular gene expression not only through classical ERalpha but also through the newly described ERbeta. These findings also demonstrate the potential for targeting of these receptors in males for prevention or treatment of heart disease.     

Relationships between estrogen receptor and estradiol-stimulated progesterone synthesis in the rabbit corpus luteum

The effects of estradiol on luteal estrogen receptor and steroidogenesis were examined on Days 9 through 11 of pseudopregnancy. In normal pseudopregnant rabbits, unoccupied cytoplasmic and total nuclear estrogen receptor were 2.6 +/- 0.4 fmol/microgram DNA and 0.4 +/- 0.1 fmol/microgram DNA, respectively, on Day 10 of pseudopregnancy. An i.v. injection of 4 micrograms of estradiol caused the translocation of cytoplasmic receptor and a 6.6-fold increment in total nuclear receptor within 15 min, which was followed by rapid processing of the nuclear receptor. Both cytoplasmic and nuclear estrogen receptor returned to normal values within 24 h, and during this period, serum progesterone did not change significantly. Withdrawal of an estradiol implant from animals on Day 9 of pseudopregnancy initiated a marked decline in serum progesterone within 24 h. Following an injection of saline or of 4 micrograms estradiol on Day 10, unoccupied cytoplasmic estrogen receptor in saline- and estradiol-injected rabbits was 1.0 +/- 0.1 fmol/microgram DNA and 1.9 +/- 0.1 fmol/microgram DNA, respectively, on Day 11 of pseudopregnancy. Associated with the increase in cytoplasmic receptor there was an increase in serum progesterone (8.2 +/- 1.5 ng/ml), in contrast to saline-injected animals in which serum progesterone continued to decline (1.6 +/- 0.4 ng/ml). Despite the significant differences in cytoplasmic receptor and in progesterone synthesis, total nuclear estrogen receptor was not different in these animals. These data suggest that in corpora lutea already secreting progesterone at high rates during midpseudopregnancy, the rapid translocation of available estrogen receptor does not cause further stimulation of progesterone synthesis. However, if steroidogenesis is first reduced experimentally, then an injection of 4 micrograms of estradiol can readily stimulate progesterone synthesis, and this stimulation is associated with an increase in cytoplasmic receptor.     

Importance of estrogen receptors in hepatic LDL receptor regulation

Treatment with pharmacological doses of estrogen is the most potent way to stimulate hepatic LDL receptor expression in vivo. The mechanism for this effect is unclear, in part because of difficulties in inducing this stimulation in vitro. A fundamental question, whether estrogen receptors (ERs) mediate this stimulation, has not been addressed. The aim of the current study was to determine the involvement of ERs in the estrogen-induced stimulation of LDL receptors. Treatment of rats with high doses of ethynylestradiol for 7 days increased the hepatic LDL receptor protein and mRNA levels four- and threefold, respectively. LDL receptor stimulation in estrogen-treated rats was not due to their reduced food intake because hepatic LDL receptor expression did not increase in rats fasted for 72 hours. Treatment with antiestrogen (tamoxifen or clomiphene) abolished the LDL receptor stimulatory effect of ethynylestradiol at both the protein and mRNA levels. Antiestrogen alone had no effect on hepatic LDL receptor expression and did not influence the strong resistance to dietary cholesterol normally present in rats. It is concluded that ERs are critically involved in the induction of hepatic LDL receptor expression by ethynylestradiol. The known role of growth hormone for the expression of hepatic ERs may therefore play a role in the modulation of the effects of estrogen on cholesterol metabolism and hepatic LDL receptors in the rat.     

Changes in the content of progesterone receptor isoforms and estrogen receptor alpha in the chick brain during embryonic development

Progesterone and estradiol participate in the regulation of several reproductive functions through interaction with intracellular progesterone receptors (PR) and estrogen receptors (ER), respectively. In this work, we determined PR and ER-alpha isoforms content in the brain of chicks of both sexes on days 8 and 13 of embryonic development as well as on the day of hatching by Western blot analysis. PR isoforms protein content increased during embryonic development in both female and male chick brain. The highest PR isoforms content was observed on the day of hatching in both sexes. Interestingly, PR-A content was higher in the brain of chick males than in that of females on day 8 of embryonic development. PR-A/PR-B ratio was higher in the brain of males than in that of females at all ages. We found two ER-alpha isoforms of 66 and 52 kDa; the content of both isoforms was higher in the brain of females than in that of males on days 8 and 13 of embryonic development. An opposite pattern of ER-alpha isoforms content was observed. In males, ER-alpha content increased during embryonic development whereas in the females it decreased during this process. These results indicate that the content of PR and ER-alpha isoforms is related to the degree of brain development in chicks, and suggest that PR and ER-alpha isoforms should exhibit sexual dimorphism in the brain of chicks during embryonic development.     

Changing estrogen and progesterone receptor patterns in breast carcinoma during the menstrual cycle and menopause

BACKGROUND: Estrogen receptor (ER) and progesterone receptor (PgR) status at the time of breast carcinoma surgery is used as a marker of both prognosis and hormone dependency to guide adjuvant therapy. The authors studied the influence of hormonal milieu at the time of surgery on ER and PgR levels. METHODS: A population of 2020 patients with breast carcinoma, including 575 premenopausal women, was analyzed. ER and PgR levels were determined by radioligand binding assays (cutoff values, 10 fmol/mg). Serum estradiol (E2), progesterone (Pg), follicle-stimulating hormone (FSH), and luteinizing hormone (LH) levels obtained on the day of surgery were used to define the menstrual cycle phase in premenopause. RESULTS: In premenopause, there was a higher proportion of ER positive (ER+) tumors in the follicular phase (62%, n = 316) than in the ovulatory phase (51%, n = 59) and the luteal phase (53%, n = 200, P = 0.03). The mean ER level was also higher in the follicular phase (30 fmol/mg) than in the ovulatory phase (20 fmol/ mg) and the luteal phase (25 fmol/mg, P < 0.001). The percentage of PgR positive (PgR+) tumors tended to be higher in the ovulatory phase (85%) than in the follicular (78%) and luteal (72%) phases (P = 0.11). The mean PgR was also higher in the ovulatory phase (177 fmol/mg) than in the follicular and luteal phases (134 and 92 fmol/mg, respectively; P < 0.001). The percentage of ER+ tumors was higher among menopausal women than among premenopausal women (67% vs. 59%, respectively; P < 0.001). Conversely, the percentage of PgR+ tumors was lower among menopausal women than among premenopausal women (65% vs. 78%, respectively; P < 0.001). In premenopause, there was a weak negative correlation between ER and E2 levels. No correlations were found between levels of ER and Pg and levels of FSH and LH or among levels of PgR and E2, Pg, and FSH and LH in premenopausal and menopausal women. CONCLUSIONS: Changes in ER and PgR levels in breast carcinoma during the menstrual cycle and menopause suggest that interpretations of hormone dependency on the basis of steroid receptor values should take into account hormonal status at the time of surgery.     

Changes in uterine estrogen and progesterone receptors during delayed implantation and early implantation in the spotted skunk

Although the exact cause(s) of embryonic diapause in the western spotted skunk and other carnivores remains unknown, it has been hypothesized that it may be due to levels of ovarian hormone secretion that are insufficient to promote a uterine environment conducive to continuous embryonic development and implantation. Immunocytochemistry was used to determine whether changes in abundance or distribution of estrogen receptors (ER) and progesterone receptors (PR) may be associated with the cessation or renewal of embryonic development. Thirty pregnant skunks were killed during delayed implantation and periimplantation periods. ER and PR were detected in luminal and glandular epithelium, endometrial stroma, vasculature, and myometrium of the uterus during the period of delayed implantation. There was a significant reduction of both ER and PR receptors during the periimplantation period. The most pronounced change was the complete loss or reduction in staining intensity for PR and ER in the luminal epithelium during the first 2-3 days after implantation. These findings suggest that the failure of skunk blastocysts to undergo continuous development and implant without a prolonged period of diapause is not the result of an insufficient number of ER or PR in the uterus. The data also indicate that renewed embryonic development and implantation is not associated with an increase in these uterine steroid receptors.