A gaschromatographic method for the measurement of small amounts of estrogens in urine

A sensitive gaschromatographic method for the measurement of estrone,estradiol and estriol in urine is described. After acid hydrolysis, extraction with ether and preliminary purification of the extracts, the three estrogens are separated and purified prior to gaschromatography. Adequate purification was achieved for estrone and estradiol by two thin-layer chromatograms,one with the free steroid and one with the acetate-derivative;for estriol with alumina chromatography of the 3-methylether derivative. The reliability of the method is discussed. Preliminary results obtained from urines of normal postmenopausal women are given

Androgens in postmenopausal breast cancer: Excretion, production and interaction with estrogens

The role of androgens and estrogens has been investigated in postmenopausal breast cancer patients. No differences were found in the urinary excretion of estrone and of estriol between 41 primary mammary cancer patients and 48 normal postmenopausal women, representative of the normal population. A significantly lower excretion of androgen metabolites (11-DOKS) was found in the patients. Because 11-DOKS in postmenopausal women arise mainly from three secretory products and estrogens are mainly derived from peripheral conversion of androstenedione to estrone, production rates of DHEA, DHEAS and androstenedione and the conversion of androstenedione to estrone were estimated. No significant difference was found between the two groups for the blood production rate of androstenedione, neither for its conversion to estrone. The urinary production rate of DHEAS was definitely lower in the selected breast cancer patients compared to normal controls. The DHEA production rate was also lower but statistical significance was not achieved. On account of these results the hypothesis was tested that DHEAS, DHEA or one of their metabolites might interfere with the binding of estradiol to its specific receptor, an essential step in its mechanism of action. From the results of anin vitro incubation study of receptors from human myometrial and mammary tumour tissue with several steroids, evidence was obtained that the estradiol binding was inhibited, in a molar concentration ratio not far beyond the physiological range, by 5-androstene-3β,17β-diol, a steroid closely related to DHEA. If these in vitro findings may be applied to in vivo conditions, it is conceivable that androstenediol is a regulating agent of estrogenic action at the cellular level. Abbreviations: E1—estrone, 3-hydroxy-l,3,5(10)-estratrien-17-one; E2—estradiol, 1,3,5(10)-estratriene-3,17β-diol; E3—estriol, 1,3,5(10)-estratriene-3,16α,17β-triol; DHEA—dehydroepiandrosterone, 3β-hydroxy-5-andro-sten-17-one; DHEAS—dehydroepiandrosterone-sulfate; DHT—dihydrotestosterone, 5α-androstane-17β-ol-3-one; T—testosterone, 4-androstene-17β-ol-3-one; A—andro-stenedione, 4-androstene-3,17-dione; Adiol—5-androstene-3β,l7β-diol; 11-DOKS—11-deoxo-17-ketosteroids, etio-cholanolone + androsterone + dehydroepiandrosteron

Estrogens in tissues: Uptake from the peripheral circulation or local production

Because intratissue levels of estrogens may be more important for the understanding of the endocrine status of an organ than are peripheral plasma levels, the concentrations of estrone and estradiol have been measured in normal and pathological breast and uterine specimens. Some breast tumors were collected in countries with differences in incidence and natural history of the disease. In other samples the subcellular distribution of the steroids was studied. Estrone levels did show much less variation than estradiol levels. Not related to estrogen receptors, estradiol levels were higher in uterine than in breast tissues. Also, the subcellular distribution observed could not be explained by changes in receptors. Malignant breast tumors of premenopausal and postmenopausal women contained similar amounts of estradiol. Unexplained large differences were found in the intratissue estradiol levels obtained in different countries

Metabolism of estradiol-17β, 5-androstene-3β,17β-diol and testosterone in human breast cancer cells in long-term culture

The human breast cancer cell line MCF-7 is able to metabolize steroids, which are added in order to study the growth rate of these cells. The following steroids: estradiol-17β, 5-androstene-3β,17β-diol and testosterone were incubated with these cells for 48 h under identical conditions used for growth-rate studies in our laboratory. The metabolites formed were identified. The conversion of estradiol-17β to estrone was 7% and no estriol was formed. The conversion of 5-androstene-3β-17β-diol to dehydroepiandrosterone was 8.1%, to testosterone 1.0% and to 5α-androstane-3β,17β-diol 3.2%. The conversion of testosterone to androstenedione was 6.5%, to etiocholanolone 0.4%, to 5α-dihydrotestosterone 0.7%, to 5-androstene-3β,17β-diol 0.5% and to androstenediol 0.2%. Some of these metabolites, due to their relative binding affinity, may have a biological effect on these cells that contain androgen and estrogen receptors

Estrogen receptors in human vaginal tissue

The presence of specific estrogen receptors could be demonstrated in vaginal tissue, obtained during operation from 38 women, age 27–75 yr. In 23 premenopausal women the receptor concentration in the vaginal tissue varied between 12 and 91 fmol/mg protein, no significant difference in the receptor level was found between the proliferative and secretory phase of the menstrual cycle, classified by endometrial histology. In 15 postmenopausal patients, the receptor level varied between 4 and 119 fmol/mg protein. In the last group a significant negative correlation (R = −0.72) was found between the vaginal estrogen receptor level and the Maturation Value of the vaginal smear; no correlations were found between the receptor level and the plasma levels of estrone, estradiol, LH, FSH and SHBG. No systematic differences in the receptor concentration in various parts of the vagina were observed. There was no correlation between the receptor level and age of the patients