Factors influencing the prognostic role of oestrogen and progesterone receptor levels in breast cancer--results of the analysis of 670 patients with 11 years of follow-up.

In the last two decades, the prognostic role of the steroid hormone receptors has been the subject of a myriad of publications. Nevertheless, its relevance after long-term follow-up is still not clear. The confusion about the prognostic value is mainly due to the difficulty in comparing analyses. Despite different study-designs and statistical approaches, oestrogen (ER) and progesterone (PR) receptors are widely accepted as prognostic factors. Data from 670 breast cancer patients with a median follow-up of 11.4 years were analysed retrospectively. ER and PR were measured by the dextran-coated charcoal (DCC) assay. To investigate the time dependence of the prognostic relevance of ER and PR, separate analyses were done for follow-up shorter and longer than 5 years. Special focus was directed at patients < or =50 and >50 years, node-negative women, in particular those without adjuvant therapy. Univariate and multivariate analyses were performed. In univariate analysis, ER and PR were associated with a significantly longer overall survival at the cut-off levels 10, 20 or 100 fmol/mg protein. The significant survival benefit occurred in the first 5 years of follow-up and remained unchanged in the following period. In the multivariate analyses, only the PR was of significant prognostic value (for PR> or =20 fmol/mg P=0.036, for PR> or =100 P=0.01, Cox analysis). In patients younger than 51 years, only PR was an independent prognosticator at the cut-off level of 100 fmol/mg protein, while in patients >50 years both hormone receptors were not significant. In N0 patients, only the PR reached long-term prognostic independence at a cut-off point of > or =100 fmol/mg (P=0.018). In addition, in the group of node-negative women < or =50 years without adjuvant therapy the PR level reached prognostic significance. The hormone receptor status was a prognostic factor only during the first 5 years of follow-up. Our data suggest that age, lymph node status, length of follow-up and probably the ER/PR assay are important for the evaluation of ER and PR as prognostic variables. In most analyses, PR appeared to be superior to ER in predicting the prognosis of primary breast cancer patients.

Dehydroepiandrosterone stimulates proliferation and gene expression in MCF-7 cells after conversion to estradiol.

Dehydroepiandrosterone (DHEA) is a mitogen for estrogen-dependent MCF-7 breast cancer cells. Our aims were to determine whether DHEA required conversion to estrogens in order to stimulate cell proliferation and estrogen-dependent gene expression. After incubation of cells with 100 nM DHEA for 4 days, estradiol was present in the medium at a concentration of approximately 200 pM. Other compounds identified were testosterone ( approximately 300 pM) and estrone. Significant stimulation of cell proliferation by 1 nM estradiol and 100 nM DHEA was observed after 38 h and 4 days of incubation, respectively, indicating the necessity of DHEA conversion. DHEA doses > or = 10 nM induced estrogen-dependent reporter gene expression in MCF-7 cells transfected with a luciferase reporter gene under the control of the estrogen response element. DHEA-dependent stimulation of proliferation and luciferase induction could be inhibited by the anti-estrogens ICI182,780 and tamoxifen, respectively, and by the aromatase inhibitor 4-hydroxyandrostenedione. An androgenic effect of DHEA on proliferation and gene expression of MCF-7 cells was not observed. We conclude that conversion of DHEA to estrogens, particularly estradiol, is required to exert a mitogenic response.

Evaluation of polyacrylamide gel as substitute for human cervical mucus in the sperm penetration test.

OBJECTIVE: To compare polyacrylamide gel as synthetic medium with human cervical mucus (CM) for the in vitro sperm-penetration test during infertility investigation. PATIENTS: One hundred sixty-nine randomly chosen couples with a median duration of infertility of 4 (range, 1 to 16) years presenting at the infertility unit of the Women's University Hospital of Heidelberg, Germany. MAIN OUTCOME MEASURES: Evaluation of sperm migration in polyacrylamide gel used in four different concentrations (1.5%, 1.6%, 1.7%, 1.8%) in the capillary tube test in parallel with CM of patients' female partners and CM of fertile donors, obtained under standardized conditions. Correlation of migration test results with outcome of semen analysis including microbial cultures and testing for local antisperm antibodies by means of the mixed antiglobulin reaction, postcoital testing, and the subsequent pregnancy rate after control for female infertility factors in a prospective study. RESULTS: Sperm ability to penetrate the synthetic medium (concerning all concentrations) correlated significantly with the penetration of human CM, although polyacrylamide proved to be a stronger barrier. Sperm velocity and duration of progressive motility were markedly reduced in polyacrylamide. Polyacrylamide results correlated with the outcome of standard sperm analyses but not with sperm antibody testing. No clear differentiation was obtained with regard to subsequent fertility (19% after 6 months), although adequate sperm migration in polyacrylamide 1.8% was significantly more frequent in the fertile group. CONCLUSIONS: In analyzing the intrinsic motility, penetration testing with polyacrylamide gel provides important information not obtained by routine sperm analysis. However, particularly with regard to immunological factors and fertility prognosis, human CM should be preferred whenever possible.

Reduced cerebrospinal fluid estradiol levels are associated with increased beta-amyloid levels in female patients with Alzheimer's disease.

Recent in-vitro studies indicate that estrogens such as 17beta-estradiol (E2) may decrease the production of beta-amyloid 1-42 (Abeta42), a peptide central for the formation of senile plaques in Alzheimer's disease (AD). To test this hypothesis in a clinical study, cerebrospinal fluid levels of E2 were compared between 30 female AD patients and 11 female patients with non-dementing diseases such as major depression and investigated with respect to beta-amyloid 1-40 and Abeta42 levels. E2 levels were significantly (P<0.05) lower in the AD group than in controls; within the AD group E2 levels were inversely correlated with Abeta42 concentrations (r=-0.36, P=0.05). This is the first clinical study providing evidence for an influence of E2 on Abeta42 metabolism in vivo. This observation corresponds to the putative beneficial effects of estrogen replacement therapy on the development and course of AD.

Serum concentrations of FSH, oestradiol, oestrone and androstenedione in normal and obese women.

The normal ranges of the concentrations of follicle stimulating hormone (FSH), oestradiol (E2), oestrone (E1), and androstenedione (A) were established in 257 healthy women. The hormone levels of 84 normal post-menopausal women were compared with those from a group of 46 post-menopausal women with severe obesity (36.3 +/- 15.2 kg over the ideal weight). The increase of the FSH concentrations during the peri- and post-menopause occurs about 4 yr earlier in the obese than in the normal women. There is no significant difference, however, between the E2 levels of the normal and obese women. In the obese women, A (P less than 0.001) and E1 (P less than 0.05) levels are significantly lower than in the normal women. A weight reduction in the obese women had no influence on the concentrations of A and E2, whereas E1 levels tended to increase. FSH levels also increased significantly during weight reduction.

Influence of severe obesity on peripheral hormone concentrations in pre- and postmenopausal women.

To investigate the influence of obesity on hormonal parameters in 186 apparently healthy women and in 176 women suffering from severe obesity the serum concentrations of FSH, LH, estrone (E1), estradiol (E2), androstenedione (A) and testosterone (T) were determined radioimmunologically. The climacteric onset of increased FSH production is 4 yr earlier (P less than 0.001) in obese than in normal women. Parallel to the rise of FSH there is a significantly premature decrease of the E1 and E2 concentrations in obese women. The typical elevation of the LH was found similar and not significantly different in the two collectives. The mean A levels are significantly lower (P less than 0.01) in obese than in normal women in all age groups. The T concentrations do not depend on the age of the women during the investigated period (41 to 60 yr) and are significantly higher (P less than 0.001) in the obese than in the normal women. There is a significant (P less than 0.001) correlation between the concentrations of A and E1 both in the obese and the normal women. An increased conversion of androgens to estrogens by adipose tissue is not revealed by the peripheral serum concentrations. Our data clearly demonstrate that in obese women the onset of ovarian insufficiency is significantly earlier than in normal women.

Weight percentile at birth. II. Prediction by endocrinological and sonographic measurements.

In a prospective study of 847 singleton pregnancies, the importance of various endocrine methods (serum estriol, HPL, SP1, beta-HCG, estradiol-17 beta, urinary estrogen excretion) and of two sonographic measurements (biparietal and thoracic diameter) for the diagnosis of growth retardation in the third trimester was studied. HPL and estriol determinations were best suited for the diagnosis of growth retardation. The thoracic diameter correlated most closely with the birthweight of the newborns. Sensitivity in relationship to growth retardation was between 17 and 35% for the HPL and estriol determinations as well as for both sonographic methods. Specificity was around 90% for these methods. The validity for all methods improved as the time of birth approached. Through the simultaneous measurement of one of the hormones and the thoracic diameter, an antepartal diagnosis of up to 50% of the hypo- and hypertrophic growth disorders was achieved. In the first two years of life a relationship between development and the HPL and estriol concentrations could be observed which was independent of the weight percentile at birth.

Maternal peripheral testosterone levels during the first half of pregnancy.

Peripheral serum testosterone levels were determined in 180 women during weeks 7 to 20 of pregnancy with a specific radioimmunoassay. After a normal pregnancy and delivery 90 serum samples were randomly selected from mothers of boys and 90 serum samples from mothers of girls. The testosterone concentrations were correlated with the sex of the fetuses. The mean testosterone level +/- S.D. in pregnant women with female fetuses was 597 +/- 167 pg. per milliliter. In pregnant women with male fetuses the testosterone concentrations were on the average significantly higher (p less than 0.01), with a mean value of 828 +/- 298 pg. per milliliter. The course of the testosterone concentrations in women with male fetuses showed an increase beginning in week 7, reaching a maximum during weeks 9 to 11, followed by a decrease until weeks 15 to 20. During weeks 9 to 11 of pregnancy fetal sex determination was possible in 28 per cent of the males and in 5 per cent of the females, with a probability of 95.5 per cent.

C3 in seminal plasma has no additional informative value in the diagnosis of infection/inflammation of the male genital tract.

The objective of this study was to determine the clinical significance of complement fraction C3 (C3c) in seminal plasma. Therefore 120 samples from randomly chosen subfertile males without signs of genital tract infection were screened for C3 and for seminal leucocytes as markers for subclinical infection/inflammation. A comprehensive semen evaluation included sperm analysis, sperm migration testing, immunocytochemical round cell differentiation to determine seminal leucocyte counts and the leucocyte ratio and semen cultures, in aliquots of the same ejaculates. C3 concentrations were significantly correlated with leucocyte counts per ml (P < 0.002) and per ejaculate (P < 0.001), and with the leucocyte ratio (P < 0.001). No association of C3 concentrations with semen quality or with the bacterial colonization of semen samples was found. The significant association with seminal leucocytes suggests that C3 might be used as an additional marker for silent male genital tract infection. In comparison with semen leucocytes, C3 screening does not reveal any further information about semen quality or infection/inflammation pathogenesis of the male genital tract.

[Therapy with gonadotropin releasing hormone agonists (zoladex) in premenopausal females with metastastic breast cancer]. / GnRH-Agonisten (Zoladex)-Therapie bei prämenopausalen Frauen mit metastasierendem Mammakarzinom.

Medicinal castration using GnRH-analogues is a new therapeutic possibility for treating metastasizing breast cancer in premenopausal women. A total of 22 premenopausal patients were included in the study reported here, all of them low-risk cases. Twenty of the 22 patients had hormone receptor-positive primary tumors. A slow-release depot form of Zoladex (ICI 118630) was used as a GnRH agonist and was administered subcutaneously (3.6 mg) at four-week intervals. The long-term administration of Zoladex brought about a significant reduction in blood FSH, estradiol, and progesterone levels within one to four weeks. In contrast, there were no detectable changes in ACTH, DHEAS, cortisol, testosterone, prolactin, or androstendion levels. Therapy-induced amenorrhea occurred in all cases. The objective remission rate achieved (complete and partial remission) was 45%. As opposed to other formulations, the use of Zoladex as a GnRH analog in depot form has significant advantages, which become particularly evident through improved compliance. With Zoladex therapy an effective drug-induced castration can be accomplished in premenopausal women. As regards its efficacy it is comparable to an ovarectomy, though with less pronounced side effects.

Endocrinology of benign breast disease in relation to the severity of radiological dysplasia.

A link between benign breast disease and hormonal imbalance has not been conclusively determined. In women suffering from severe radiological dysplasia, it was found to be correlated with slightly raised serum hormone binding globulin (SHBG) binding capacity. SHBG is an oestrogen-sensitive protein, which may be a marker of enhanced oestrogen bioactivity. Such abnormality was not, however, detected by the measurement of peripheral steroid hormones.

[Proliferation index, axillary lymph node status, hormone receptors and age as prognostic factors in primary breast cancer]. / Proliferations-Index, axillärer Lymphknotenstatus, Hormonrezeptoren und Alter als Prognosefaktoren beim primären Mammakarzinom.

A total of 233 primary carcinomas of the breast (n = 68 nodal negative and n = 165 nodal positive) of 77 woman patients of less than 50 years of age and pf 156 women patients of 50 years of age or older were examined for the prognostic significance of a proliferation index (in vitro chemoresistance assay). The prognostic significance of the proliferation index (high vs low proliferation) is compared with other prognostic factors that have already been in clinical use (age, axillary lymph node status, oestrogen and progesterone receptor status). The total median observation period was 4.8 years (3-9.1 years). Determination of the proliferation index proved to be the most important prognostic factor both for premenopausal and postmenopausal, nodal negative and nodal positive primary tumours when assessing the total survival time and the time that remains free from recurrences. A simultaneous assessment of low proliferation index and positive hormone receptor as a so-called low-risk situation shows in comparison to a high-risk situation (high proliferation index, negative receptor status) the most pronounced differences. For premenopausal women (younger than 50 years of age) the hormone receptor status has no prognostic significance as far as our group of patients is concerned, whereas for women of 50 years of age or older the receptor status is of great prognostic relevance. Besides the lymph node status, the proliferation index shows in a Cox regression model the greatest prognostic importance for the further course of a primary carcinoma of the breast.(ABSTRACT TRUNCATED AT 250 WORDS)

[The effects of hormone replacement therapy on ovarian function in premenopausal women after hysterectomy]. / Auswirkungen einer hormonellen Substitutionstherapie bei hysterektomierten prämenopausalen Frauen auf die ovarielle Funktion.

OBJECTIVE: Examining the consequences of temporary postoperative hormone replacement therapy following hysterectomy for the function of the ovaries and the subjective well-being of women. MATERIAL AND METHODS: Hormone profiles (Estradiol, FSH, LH, Testosterone, DHEA) and typical estradiol deficiency phenomena were investigated prospectively in premenopausal hysterectomized women with intact ovaries. Group 1 (n = 21) was replaced transdermally following surgery for 3 weeks with estradiol patch 0.05 mg daily. Group 2 (n = 21) got no hormones. RESULTS: Group 1 had a remarkable decrease of estradiol after 10 days to 59% and after 6 weeks to 71% of the starting point. Gonadotropins showed an increase in this group. In group 2 without replacement there was only a small decrease of estradiol after 10 days and after 6 weeks the level was higher than before hysterectomy. Testosterone also decreased in group 1 to 64% of the level before surgery after 6 weeks, whereas in the comparing group it was 87%. On the other hand in group 1 only 2 of 21 women, but 10 of 21 in group 2 showed climacteric-like symptoms. CONCLUSIONS: HRT over 3 weeks induces ovarian suppression, which is still seen 6 weeks after hysterectomy. But hormonally treated women have clearly less subjective complaints.

Distribution of estrogen and progesterone receptors on primary tumor and lymph nodes in individual patients with breast cancer.

Primary breast cancer tissue and lymph nodes were obtained from 55 patients, Histologically, 34 of these patients had positive and 21 negative lymph nodes. Estrogen receptors (ER) and progesterone receptors (PR) were determined by a dextran-coated charcoal assay. The tumor tissue was ER positive in 58% of the cases and PR positive in 34%. The malignant lymph nodes were ER positive in 56% and PR positive in 24%. ER in 14% and PR in 5% of the benign lymph nodes could be detected. The primary tumor tissue and the corresponding malignant lymph nodes showed an identical ER and PR status, i.e. both tumor sites were receptor positive or both receptor negative, in 68 and 74%, respectively. However, 21% of the patients had receptor-positive tumors but receptor-negative lymph nodes. Receptor-positive lymph nodes in combination with receptor-negative tumors occurred in only 11% for ER and 6% for PR. These data show that receptor-positive malignant lymph nodes mostly display the same receptors status as the corresponding primary tumor, whereas receptor-negative lymph nodes may be combined with receptor-positive tumors.

Steroids in human myometrium and peripheral blood during the menstrual cycle.

Blood samples were collected prior to vaginal hysterectomy from nine women in the proliferative phase of the cycle and from 15 women in the secretory phase, and from each uterus five grams of the fundus were obtained. The concentrations of progesterone (P), 20alpha-dihydroprogesterone (20alpha-DHP), estradiol-17beta (E2), and estriol (E3) were determined in serum and in the tissue with the use of specific radioimmunoassays. The concentrations of P and 20alpha-DHP in serum and myometrium were significantly higher in the secretory than in the proliferative phase. Furthermore, the concentration of P in myometrium was significantly higher than in serum in the secretory phase, indicating a specific uptake of P by the myometrium. The concentration of E2 was about 10 times higher in the myometrium than in the serum during both phases of the cycle and appeared to be independent of the serum levels. This indicates a specific but limited uptake of E2 by the myometrium. The E3 levels in the tissue showed no significant differences during the menstrual cycle, whereas in the serum the E3 concentrations were higher in the secretory than in the proliferative phase of the cycle.

Iliac crest needle biopsy as a method for determining estrogen receptors in bone metastases from breast cancer.

The purpose of our study was to clarify whether the amount of tissue extracted by means of iliac crest needle biopsy (ICNB) would suffice for a quantitative determination of estrogen receptors (ER) in bone metastases, and to see if the method currently used for determining ER in primary tumors could also be successfully utilized in ICNB. 22 of 31 breast cancer patients examined could be evaluated. ER was positive in 7 (31.8%). Reasonable data are to be expected when the biopsy weight exceeds 0.1 g. Our study confirms that the necessary amount of tissue for ER analysis can indeed be extracted by ICNB. Our results justify further studies on a larger group of patients, since we cannot make conclusive statements concerning the value of this method for predicting the ultimate success of an endocrine treatment.

[Endocrinology of the climacteric]. / Endokrinologie des Klimakteriums.

The endocrinology of the menopause is determined by the decrease of estrogens. In 250 healthy patients there was an observed decrease of estrone to 35 pg/ml and of estradiol to 10 pg/ml serum between the 49th and 54th year of age. Thereafter no further decline occurred, because the androgen precursors of the estrogens were secreted by the adrenal cortex until advanced age. In a period of 30 years there was a decrease of the androgen levels of only 25%. In 200 patients with severe obesity we found no differences in peripheral steroid concentrations, but the SHBC concentrations were significantly lowered; therefore, the amount of free active estradiol and the effect on the target organ is higher in these patients.

Stereologic correlates of steroid receptor concentration in invasive ductal breast cancer.

The hypothesis was tested that morphometric parameters of tumor cell nuclei correlate with the steroid receptor concentration in mammary carcinoma. In 50 consecutive mastectomy specimens with a diagnosis of invasive ductal cancer in which estrogen receptor (ER) and progesterone receptor (PR) concentrations had been assayed quantitatively, morphometric measurements were performed on four visual fields of two sections per case. The fields were sampled from the most cellular regions of the tumor. The number of tumor cell nuclear profiles per tissue area, the nuclear profile area and the long and short nuclear profile axes and their ratios were measured with a semiautomatic image analysis system. Estimates of the number of tumor cell nuclei per tissue volume (Nv) and of the mean tumor cell nuclear volume (V) were obtained by standard stereologic techniques. Association between the morphometric and biochemical parameters was tested by Spearman's rank correlation coefficient. Nv correlated positively with the steroid receptor concentration whereas V correlated negatively with both ER and PR concentrations. A correlation of the receptor concentrations to the standard deviation of the nuclear area or the mean ratio of the nuclear axes could not be demonstrated. These results suggest that receptor-rich tumors have a large number of small tumor cell nuclei whereas receptor-poor tumors have a small number of large tumor cell nuclei per tissue volume in the actively proliferating, highly cellular regions. These differences are not accompanied by significant changes in nuclear size variability or nuclear shape.