Concentrations of estrone, estradiol, and estrone sulfate and evaluation of sulfatase and aromatase activities in pre- and postmenopausal breast cancer patients.

This report concerns the evaluation of various estrogens, estrone (El), estradiol (E2), and estrone sulfate (E1S), as well as E1S-sulfatase and aromatase activities in pre- and postmenopausal women with breast cancer. The levels (in picomoles per g; mean +/- SEM) of the various estrogens in the breast tissue from premenopausal patients (n = 11) are: El, 1.4 +/- 0.5; E2, 1.2 +/- 0.6; and E1S, 1.2 +/- 0.3. In postmenopausal patients (n = 23), the values are, respectively, 1.0 +/- 0.4, 1.4 +/- 0.7, and 3.3 +/- 1.9. These concentrations of estrogens in the tumors of postmenopausal patients are significantly higher than those found in plasma. The activity of E1S-sulfatase in both pre- and postmenopausal patients was 50-200 times higher than that of aromatase. E1S-sulfatase and aromatase activities are significantly higher in post-menopausal than in cycling patients. It is concluded that despite the low levels of circulating estrogens in postmenopausal patients, the tissue concentrations of these steroids are several-fold higher than those in plasma, suggesting tumor accumulation of these estrogens. The physiopathology and clinical significance of these high levels of the various estrogens (E1, E2, and E1S) as well as sulfatase and aromatase activities in postmenopausal patients with breast cancer is yet to be explored.

Estrone sulfate-sulfatase and 17 beta-hydroxysteroid dehydrogenase activities: a hypothesis for their role in the evolution of human breast cancer from hormone-dependence to hormone-independence.

The evaluation of estrogens (estrone, estradiol, and their sulfates) in the breast tissue of post-menopausal patients with breast cancer indicates high levels, particularly of estrone sulfate (E1S) which is 15-25 times higher than in the plasma. Breast cancer tissue contains the enzymes necessary for local synthesis of estradiol and it was demonstrated that, despite the presence of the sulfatase and its messenger in hormone-dependent and hormone-independent breast cancer cells, this enzyme operates particularly in hormone-dependent cells. Different progestins: Nomegestrol acetate, Promegestone, progesterone, as well as Danazol, can block the conversion of E1S to E2 very strongly in hormone-dependent breast cancer cells. The last step in the formation of estradiol is the conversion of E1 to this estrogen by the action of 17 beta-hydroxysteroid dehydrogenase. This activity is preferentially in the reductive direction (formation of E2) in hormone-dependent cells, but oxidative (E2-->E1) in hormone-independent cells. Using intact hormone-dependent cells it was observed that Nomegestrol acetate can block the conversion of E1 to E2. It is concluded, firstly, that in addition to ER mutants other factors are involved in the transformation of hormone-dependent breast cancer to hormone-independent, this concerns the enzymatic activity in the formation of E2; it is suggested that stimulatory or repressive factor(s) involved in the enzyme activity are implicated as the cancer evolves to hormone-independence; secondly, different drugs can block the conversion of E1S to E2. Clinical trials of these "anti-enzyme" substances in breast cancer patients could be the next step to investigate new therapeutic possibilities for this disease.

[Antiovulatory action of chlormadinone acetate]. / Action anti-ovulatoire de l'acétate de chlormadinone.

Antiovulatory action of chlormadinone acetate (5 mg twice daily from day 7 to day 25) has been assessed in 6 healthy volunteers by daily determination of plasma FSH, LH, estradiol and progesterone. Hormonal profiles during the second treated cycle show that preovulatory gonadotropin surge is blunted and that no significant progesterone secretion occurs. Estradiol production is variable up to the middle of the cycle, and then homogeneously low normal. Menstrual cyclicity is respected and ovarian function is restored during the first cycle after treatment disruption.

[Antiovulatory action of chlormadinone acetate]. / Action anti-ovulatoire de l'acetate de chlormadinone.

PIP: The antiovulatory action of chlormadinone acetate was studied in six healthy volunteers who were given two daily doses of 5 mg each from the seventh to the twenty-fifth cycle day. Chlormadinone acetate is of potential interest as a contraceptive method for the small group of women with contraindications to synthetic estrogens and norsteroid progestins for whom no other methods are acceptable. The observation period included four cycles: a control cycle before treatment during which ovulation was confirmed, two treatment cycles, and an observation cycle after treatment. Antiovulatory action was assessed by daily determination of plasma FSH, LH, estradiol, and progesterone levels. The results confirmed the antiovulatory action of chlormadinone acetate and revealed no clinical signs of functional ovarian cysts or premenstrual syndrome. In all cases, the LH surge observed in the control cycle was blunted by chlormadinone acetate, while the base gonadotropin levels were not modified. Hormonal profiles in the second treatment cycle showed no preovulatory gonadotropin surge and no significant progesterone secretion. Estradiol production was variable through midcycle and then low normal in all subjects. In the last cycle week, the estradiol level was below 50 pg/ml in two cases and between 50 and 100 pg/ml in the others. In five cases out of six, the level of progesterone was 1 ng/ml or lower. In the sixth case, the data were incompatible with ovulation. Cycle tolerance was good and ovarian function returned during the first posttreatment cycle. No significant variation in weight or blood pressure was observed at the end of the second treatment cycle.^ieng

Studies on the influence of gonadotrophin levels in the early follicular phase on the ovarian response to stimulation.

The gonadotrophic regulation of folliculogenesis has been extensively investigated but little attention has been paid to the influence of early follicular phase levels of endogenous FSH and the FSH/LH ratio when planning ovulation stimulation therapy for IVF. The influence of these factors was investigated in the three studies reported in this paper. A fixed schedule of ovulation stimulation therapy which employed standard treatment regimens, irrespective of the ovarian response, was used to eliminate variation due to treatment factors. Cycles were pretreated with an oestrogen-progestogen contraceptive pill or a progestogen (norethisterone). It was found that both oestrogen-progestogen and progestogen alone decreased the plasma FSH level, although the FSH/LH ratio was significantly reduced only by oestrogen-progestogens. In clinical IVF studies, oestrogen-progestogen pretreatment was associated with a significant reduction in the preovulatory concentration of oestradiol in plasma and the number of aspirated follicles, compared to norethisterone. The administration of FSH for 2 days following oestrogen-progestogen pretreatment and prior to the fixed schedule of ovulation stimulation normalized ovarian steroidogenesis and follicular development. Early follicular phase supplementation with FSH had no influence on progestogen pretreated cycles. The final experiment investigated the influence of FSH/LH levels in the early follicular phase on the outcome of ovarian stimulation. The preovulatory oestradiol concentration was reduced when baseline FSH/LH levels were low compared with when these values were high. Administration of FSH for 2 days in the early follicular phase improved the preovulatory level of oestradiol when baseline FSH/LH was low but had no effect when baseline FSH/LH levels were high.(ABSTRACT TRUNCATED AT 250 WORDS)

Normal and adenomatous human pituitaries secrete thyrotropin-releasing hormone in vitro: modulation by dopamine, haloperidol, and somatostatin.

TRH is present in human normal pituitaries and in pituitary adenomas. In this study we demonstrated that the same tissues can release TRH in vitro. Fragments from seven normal pituitaries (10-15 mg/syringe) and dispersed cells from eight prolactinomas, four GH-secreting and two nonsecreting adenomas (1-3 x 10(6) cells/syringe) were perifused using a Krebs-Ringer culture medium. After 1 h of equilibration the perifusion medium was collected every 2 min (1 mL/fraction) for 3 h. TRH, PRL, and GH were measured by RIA under basal conditions and in the presence of 10(-10) to 10(-6) mol/L dopamine (DA), alone or concomitant with haloperidol, or in the presence of 10(-10) or 10(-6) mol/L somatostatin. Both normal pituitary fragments and pituitary adenomatous cells (from all types of adenomas studied) spontaneously released TRH in vitro. TRH was detected in the perifusion medium either immediately after the end of the equilibration period or 30-60 min later. The molecular identity of TRH was assessed by high pressure liquid chromatography. There was no difference in the profile and the rate of TRH secretion between normal and tumoral tissues, and no correlation was found between the level of TRH release and that of PRL or GH secretion. DA stimulated TRH release from normal pituitaries and from PRL- and GH-secreting adenomas at doses as low as 10(-10) mol/L. A concomitant decrease in PRL and GH release was observed from adenomatous cells and in one case of normal tissue. Haloperidol (10(-7) mol/L) antagonized the effect of 10(-8) mol/L DA on both TRH and PRL secretion in normal pituitary and in prolactinomas. DA had no effect on TRH release from two nonsecreting tumors. The amounts of TRH released during 1 h of perifusion were 60-1640 pg/2 mg wet wt tissue in normal pituitaries and 54-2174 pg/10(6) cells in adenomas; these values were very high compared to those precedently reported within the tissues. These results indicate that pituitary cells can release TRH in vitro and suggest that TRH might be synthesized in situ. We suggest that TRH could act on pituitary hormone secretion and/or cell proliferation via a paracrine and/or an autocrine mechanism.

[hCG and its subunits as tumor markers]. / hCG et ses sous-unités comme marqueurs tumoraux.

hCG is known to be produced by trophoblastic cells as well as by many normal tissues and malignancies. hCG is considered as a reliable marker for gonadal and extra gonadal choriocarcinomas; moreover it has been evidenced by immunohistochemical techniques in other malignancies. In this study, hCG and/or its alpha and beta subunits were detected in 73 males. Intact hCG was detected in 48 patients with gonadal tumors (n = 37) or extragonadal malignancies (n = 11), with or without free alpha or beta subunits. In the other 25 patients, native hCG was not detected but there were significant amounts of free beta subunit in 8 germ cells tumors and 17 extragonadal tumors. The production of free beta chains is correlated with a severe evolution in extragonadal tumors. hCG should be considered as a good marker for many tumors, but it is absolutely necessary to evaluate intact hCG as well as free beta chains.

[Neurohormones coming from the normal and tumoral human anterior pituitary. Secretion and regulation in vitro]. / Neurohormones provenant de l'antéhypophyse humaine normale et tumorale. Sécrétion et régulation in vitro.

Several neuropeptides, classically associated with the hypothalamus have been found in the anterior pituitary and their local synthesis has been hypothesized. Using normal and tumoral human pituitaries we found in the tissue itself different neuropeptides (TRH, SRIH, GHRH) and dopamine in variable quantities according to the nature of the tissue. They were all present in normal pituitaries while only stimulatory neurohormones like TRH and GHRH were found in tumoral tissue implying an imbalance between the stimulatory and inhibitory control of hypophyseal hormones (PRL and GH) in pituitary adenomas. Fragments from normal pituitaries and dispersed cells from GH, PRL and nonsecreting adenomas, were perifused for 4 hours in a Krebs-Ringer medium collected every 2 min and GH, PRL, TRH, GHRH and SRIH were measured by RIA under basal conditions and in the presence of 10(-6) mol/L DA, TRH or SRIH. Neuropeptides and DA were characterized by HPLC. Both normal and tumoral pituitaries released TRH, SRIH and GHRH in large amounts suggesting their local synthesis. There was an in situ regulation between SRIH and GH as their secretion profile was negatively correlated, GH secretion decreasing while SRIH secretion was increasing. Moreover the release of TRH was stimulated 5 to 20 folds by DA, while PRL decreased at the same time. Pulses of TRH and SRIH had differential effects on GHRH and SRIH release according to the nature of the tissue as TRH stimulated SRIH release from normal pituitary while it inhibited SRIH release from adenoma. These results indicate that anterior pituitary cells can release neuropeptides which are probably endogenously synthesized and have a local regulation.(ABSTRACT TRUNCATED AT 250 WORDS)

[Release of thyrotropin releasing hormone (TRH) from human prolactin-secreting pituitary adenoma cells. Modulation by dopamine]. / Libération de thyrolibérine (TRH) par les cellules adénomateuses hypophysaires humaines à prolactine. Modulation par la dopamine.

We found, by radioimmunoassay, that thyrotropin-releasing-hormone (TRH) was present in human prolactin (PRL)-secreting adenomas (mean: 89 +/- 45 (SEM) fmol/mg proteins) and was released by perifused adenomatous cells at levels varying from 5 to 60 fmol/10(6) cell/2 min. TRH release was increased in the presence of dopamine (DA) 10(-6) M but was not modified by the presence of somatostatin (SRIH) 10(-6) M.

Gonadotropin and alpha-subunit secretion during long term pituitary suppression by D-Trp6-luteinizing hormone-releasing hormone microcapsules as treatment of precocious puberty.

Short term treatment with GnRH agonists has been reported to increase plasma gonadotropin alpha-subunit (Gn alpha) levels while decreasing plasma immunoreactive LH (IR-LH) levels. In this study we examined the effect of D-Trp6-LHRH (LHRH-A) in microcapsules (60 micrograms/kg, im, every 28 days for 1 yr) in 13 girls suffering from precocious puberty. Plasma IR-Gn alpha was measured by RIA; plasma IR-LH and IR-FSH were measured by both polyclonal RIAs and monoclonal immunoradiometric assays (IRMA). Before treatment, basal IR-LH and IR-FSH levels and peak responses to LHRH measured by both RIA and IRMA were similar, and the Gn alpha response paralleled that of LH. After the first injection of LHRH-A, RIA LH levels were significantly higher than pretreatment levels until day 21, while IRMA LH levels transiently increased, but returned to pretreatment levels by day 7 and became lower thereafter (P less than 0.005). Plasma IR-Gn alpha levels increased from days 3-21 (P less than 0.05). After 1.5 months of treatment, basal RIA LH levels remained detectable and not different from pretreatment levels; IRMA LH levels were very low. The mean RIA and IRMA LH responses to LHRH were decreased at 1.5 and 12 months (P less than 0.01). Basal plasma RIA and IRMA FSH levels were similar during treatment (P greater than 0.05) and significantly lower than pretreatment values (P less than 0.01). The mean RIA and IRMA FSH responses to LHRH decreased significantly at 1.5 months (P less than 0.001). After 12 months, both RIA and IRMA FSH responses were increased, but IRMA values were significantly lower than RIA values. A sustained increase in basal Gn alpha values occurred, but there was a tendency for the peak levels after LHRH treatment to decrease, becoming significantly lower than pretreatment peak levels after 1 yr. The chromatographic analysis on Sephadex G-100 of a pool of plasma samples collected during a LHRH test in three children treated for 6 months indicated that IR-Gn alpha coeluted with [125I]Gn alpha. The large discrepancy between RIA and IRMA LH values suggests the secretion of unusual LH molecules which are recognized by RIA but not by IRMA. The sustained release of large amounts of IR-Gn alpha indicates dissociated effects of LHRH-A on alpha- and beta-subunit secretion by the gonadotrophs. The sustained response of Gn alpha to LHRH demonstrates that gonadotroph cell LHRH receptors are still responsive to LHRH during treatment with a LHRH agonist.

Interrelationship of plasma and urinary luteinizing hormone preovulatory surge.

The only reliable method of predicting spontaneous ovulation relies on the detection of the preovulatory luteinizing hormone (LH) surge in urine or plasma. The efficiency of the detection by means of plasma LH radioimmunoassay, urine LH radioimmunoassay or urine LH agglutination inhibition immunoassay were compared in 33 patients. The detection of the onset of LH surge was simultaneous in plasma and urine in only 11 cases. In two thirds of the patients, the urine LH surge onset is delayed by 3 to 21 h as compared with plasma LH surge onset. In some of these cases the oocyte would probably be missed if the laparoscopy had been scheduled according to urine data.

[Evaluation of plasma tumor markers in germinal tumors of the testis]. / Evaluation des marqueurs tumoraux plasmatiques dans les tumeurs germinales du testicule.

Pregnancy specific beta 1 glycoprotein (PS beta 1G), human choriogonadotropin (hCG), alpha-foetoprotein (AFP) and carcinoembryonic antigen (CEA) were measured simultaneously in 58 patients with a progressive germinal testicular tumour, and in 20 patients in complete remission. All markers were negative in all the cases in remission, and in 1 case of progressive teratoma. The specificity of the association of these 4 markers was 76 p. 100, higher than any of them measured alone. There was no correlation between individual markers with respect to histological classification. The highest diagnostic specificity was obtained with hCG in all types of tumours. PS beta 1G was associated with hCG but with a lower specificity. AFP is mainly related to embryonic carcinoma but may be raised in all forms of tumour, except seminomas. CEA was positive less often, irrespective of the type of tumour, and never alone. The value of measuring these tumour markers is double: on the one hand it allows better characterisation of the tumour, and on the other, the clinical course and efficacy of treatment can be followed-up more closely and tumour recurrence may be diagnosed at an early stage.

[Tumors secreting choriogonadotropin in children]. / Les tumeurs de l'enfant sécrétant de la choriogonadotropine.

Fourteen cases of sexual precocity due to choriogonadotropin (hCG) secreting tumors are reported. One pineal and 4 gonadal tumors were observed in 5 girls, whilst 2 supra-sellar, 4 mediastinal, 1 gonadal and 2 hepatic tumors were observed in 9 boys. In all children, immunoreactive plasma LH was high (4 to 30 mIU/ml) but non-responsive to LH-RH, due to the presence of hCG (15 to 6,000 mIU/ml). In the 4 boys with mediastinal tumors the karyotype was 47,XXY. In girls, hCG secreting tumors are a uncommon cause of sexual precocity (less than 1 p. 100). In boys their incidence seems to be the same as the incidence of idiopathic precocious puberty, that is 21 p. 100 of sexual precocities due to androgenic secretion of testicular origin.

[Plasma and intrafollicular hormone profiles in the late preovulatory phase. I. Spontaneous cycles]. / Profil hormonal plasmatique et intrafolliculaire au cours de la période préovulatoire tardive. I. Cycles spontanés.

The hormone state of development in the late preovulatory phase is described in its relationship to the start of the LH surge which occurs 37-39 hours before ovulation. No precise relationship has been found between the oestradiol (E2) peak in the plasma and that which occurs before ovulation. All the same, the succession of hormone events that have been observed in 77 spontaneous cycles is the following: the level of E2 suddenly rises between 35 and 30 hours before the ovulatory discharge and this rise is immediately followed by a rise in the level of LH. Then the plasma concentration of 17-hydroxyprogesterone (17-OHP) increases 5 hours before the LH surge. In the preovulatory follicle 5 hours after the start of the LH surge, a drop in the level of E2 and a rise in the level of progesterone is noted. Concentrations of androgens only lessen in the last 12 hours before ovulation whereas the level of 17-OHP stays stable. We have thus been able to work out the sequence of plasma hormone phenomena that occur in determining whether a preovulatory gonadotrophic surge will occur. We have described the hormone levels in follicular fluid immediately before spontaneous ovulation. These findings will be able to serve as a reference to evaluate the quality of the late preovulatory phase in abnormal cycles or in cycles that have been induced.

Pituitary and ovarian dysfunctions in women on haemodialysis.

28 female uraemic patients treated by chronic haemodialysis were studied. Of these patients, 6 had regular cycles, 6 irregular cycles, 7 were amenorrhoeic and 9 post-menopausal. Plasma gonadotropins (FSH and LH) were determined in all patients. Plasma concentrations of oestradiol and progesterone were determined only when of possible interest. LRH stimulation test was performed in 5 amenorrhoeic patients. In 19 pre-menopausal women baseline plasma FSH values were always normal, but LH concentrations were above normal in 5. In all 9 post-menopausal women both FSH and LH were markedly higher than in the pre-menopausal patients., either healthy or uraemic. Persistent oestrogen activity was found in the 6 regularly menstruating women as well as in the 5 women with oligomenorrhoea. Luteal phase plasma progesterone was in the low normal range in 3 patients and was markedly depressed in the others. A prolonged rise in LH and to a lesser extent in FSH was observed in 4 of the 5 patients studied after 100 micrograms of LRH were administered intravenously. These results suggest that the defect underlying gonadal dysfunction in uraemic women treated by chronic haemodialysis is mainly of suprahypophyseal origin.