Gianpiero Gaidano

The receptor-mediated action of sex steroid binding protein (SBP, SHBG): Accumulation of cAMP in MCF-7 cells under SBP and estradiol treatment

The interaction of sex steroid binding protein (SBP) with its specific receptor in MCF-7 cell (estrogen-sensitive human breast cancer cells), followed by the binding of estradiol (E2) to the complex SBP-receptor, induced a significant accumulation of intracellular cAMP. SBP alone as well as E2 alone did not elicit any modification of the nucleotide. The maximal increase in cAMP was observed with 1 nM SBP + 1 nM E2. Increasing doses of both SBP and E2, even raising cAMP levels with respect to basal, did not give any higher response. Both testosterone and dihydrotestosterone, used instead of E2, were not able to induce any significant modification of cAMP. E2-induced MCF-7 cell proliferation was significantly reduced by 8Br-cAMP. MDA-MB 231 cells (estrogen-insensitive breast cancer cells) were not shown to bind SBP, or to respond to SBP + E2 as far as both their proliferation and cAMP content are concerned. In summary, the present study provides evidence that the SBP receptor is part of the G-protein receptor family, and that SBP can act as modulator of E2 action at cell site through the second messenger cAMP.

Sex steroid binding protein (SBP) receptors in estrogen sensitive tissues

Since the discovery of a specific membrane binding site for sex steroid binding protein (SBP) in human decidual endometrium and in hyperplastic prostate numerous speculations have been raised on the existence of an additional non-receptor-mediated system for steroid hormone action. In the present work SBP cell membrane binding was investigated in human estrogen target tissues other than those previously studied either in the absence of steroids or in the presence of varying amounts (10(-10)-10(-6) M) of estradiol, testosterone and dihydrotestosterone, respectively. Plasma membranes obtained by differential centrifugation from homogenized samples of pre-menopausal endometrium, endometrium adenocarcinoma, normal liver and post-menopausal breast showed a specific binding of highly purified [125I]SBP: a major displacement of labeled SBP was elicited by radioinert SBP, while no significant displacement occurred when other human plasma proteins were used as cold competitors (molar excess ranging 500-10,000-fold). A specific, time-dependent binding of [125I]SBP was also observed in MCF-7 and in Hep-G2 cell lines. The different patterns of specific binding, observed in membranes from different tissues when SBP was liganded with different sex steroid molecules, leads us to consider the tissue individuality of the receptor as a further entity in the membrane recognition system for SBP.

Smoking Effects on the Hormonal Balance of Fertile Women

We evaluated serum pituitary hormones (prolactin, follicle-stimulating hormone, luteinizing hormone), gonadal hormones (estrone, estradiol, progesterone), sex steroid binding protein (SBP) and urine estrogens in 684 healthy fertile women, subdivided into smokers (n = 237) and nonsmokers (n = 447). The aim of the work was to elucidate whether smoking habits can affect hormonal balance. Smoking interference of estrogen metabolism has been postulated, but no unequivocal data have been reported. A protective role against breast cancer has even been suggested on the basis of a reduced estrogenic activity found in smokers. Our data showed a considerable interference of smoking on PRL secretion, probably related to a direct inhibiting activity of nicotine. Estrogen catabolism could also be involved, and a catabolic shift of 16 alpha-hydroxylation in favour of 2 alpha-hydroxylated catabolites, via the hepatic cytochrome P-450 system could be hypothesized.

The receptor for human sex steroid binding protein (SBP) is expressed on membranes of neoplastic endometrium.

Sex steroid binding protein (SBP) receptor was detected on cell membranes obtained from human endometrium adenocarcinoma. The binding of SBP was proved to be highly specific, saturable, and at high affinity. It was, additionally, shown to occur at two sites at different affinities, as previously described for other human tissues. SBP was, therefore, demonstrated to recognized a specific receptor on endometrium adenocarcinoma membranes. The effect of steroid hormones on SBP-receptor interaction was also evaluated. Both dihydrotestosterone and estradiol were shown to inhibit the binding of SBP to its specific receptor on neoplastic membranes. Testosterone at a dose of 10(-9) M was shown not to interfere to a significant extent with SBP-receptor binding. The sensitivity for estradiol we had previously observed in normal premenopausal endometrium was completely lost in postmenopausal neoplastic tissue. These observations suggest that the SBP-membrane recognition system is still present in neoplastic postmenopausal endometrium, but it has been modified either by the postmenopausal endogenous milieu or by the neoplastic transformation.

Hormonal and clinic evaluation of patients with moderate body hair growth

Plasma prolactin (PRL), gonadotropins (FSH, LH), estradiol-17 beta (E2), progesterone (P), total testosterone (T), sex steroid binding protein (SBP), T/SBP index, cortisol (F), 17-OH-progesterone (17OH-P), dehydroepiandrosterone sulphate (DHEA-S) and androstenedione (A), were measured in 50 fertile non-obese women presenting with moderate body hair growth and in 30 matched controls. DHEA-S and PRL were significantly higher (P < 0.002, P < 0.001, respectively) and SBP was lower (P < 0.001) in patients than in controls. Regression analyses showed that PRL levels were independent of the other parameters, while a negative correlation was found between DHEA-S and SBP values. Since the decision to treat a woman with mild body hair growth is usually a clinical one, PRL behaviour has to be taken into account before deciding the type of treatment. Clinical improvement was observed in subjects treated with ethynylestradiol plus desogestrel or plus cyproterone acetate, so as to produce an increase in SBP rather than a decrease in DHEA-S.

Behaviour of serum α-fetoprotein in acute hepatitis

Summaryα-FP was determined by radioimmunoassay in the serum of 28 normal subjects aged between 21 and 40 and in the serum of 27 male subjects aged between 19 and 49 with acute hepatitis. The measurements made on the hepatitis cases were systematically repeated during the course of the disease from the 5th day after its clinical appearance to the 30th–45th day; at the same time the behaviour of GPT, alkaline phosphatase and bilirubinemia was also studied. In normal adult subjects, α-FP was constantly present with a mean value of 1.37±0.67 in males and 1.10±0.63 in females; no statistically significant difference was found between the two sexes. In the hepatitis cases α-FP was found in significantly higher quantities than the normal mean values; it increased progressively to reach amaximum on about the 15th–20th day from the clinical onset, and then decreased quite rapidly remaining, however, at a level higher than normal for the whole observation period. The findings confirm the persistence of the α-FP synthesis in the normal adult subject and reveal an increase in this synthesis in the course of acute hepatitis, possibly linked to the process of hepatic regeneration, but without this synthesis being correlated with the behaviour of the other serum parameters (GPT, alkaline phosphatase, bilirubin).