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178. Specific binding of testosterone (T) and 5α-dihydro-testosterone (5α-DHT) to plasma proteins in human males with benign prostatic hypertrophy, prostatic carcinoma, after hypophysectomy and after orchidectomy
33x
Abstracts
tation. equilibrium dissociation constants of the hormonecomplexes were determined in the clone M1, : K, = 0.49 + 0.04 nM i’or the androstandlone binding to the A-receptor. and 0.41 f 0.04nM for estradiol binding to the E-receptor. The concentration of the hormone receptors in the cytosol of the clone cells were 79 & 0.04 pM/mg protein for the A-receptor and 49 i 0.02 pMimg protein for the E-receptor. The MI, clone cells contained approx. 20 times more A-receptor and approx. IO times more E-receptor than the primary culture cells.
176. Steroid hormone receptors in human renal carcinoma FANESTIL, D. D., VAUGHN. D. A. and LUDENS. J. H.. Department of Medicine, University of California. San Diego. La Jolla, California. U.S.A. Slices (0.5 mm thick) from 5 human renal adenocarcinomas and from adjacent normal renal tissue were incubated in Krebs-Ringer solution for 60 m at 37’ C in the presence of 6 x 10m9 M H”-aldosterone. H3-testosterone or H3-progesterone (in 4 cases) both in the absence and in the presence of 6 x IO-’ M non-radioactive steroid of the respective type. Cytosol (105.000 pg supernatant) and purified nuclei (PN) obtained by sedimenting the 6OOpg pellet through 2.2 M sucrose were prepared from ail slices. The CH,Cl,-extractable H3-steroid bound to protein was determined in cytosol after Bio-gel filtration and in PN. The amount of HA-steroid displaced by the lOO-fold excess of non-radioactive steroid was taken as “specific binding”. In general, the tumour cytosol contained Z-3 times more specifically bound steroid than normal cytosol. However, the nuclear specific binding was more variable. The normal and tumour nuclei contained 3.7 + 3.5 (mean i SEM) and 0.5 + 0.3 x IO ” mot aldosterone/mg protein; for testosterone the respective values were 1.05 i 3.9 and 15.7 k 8.1; for progesterone I.70 + .86 and 18.7 + 17.6. Thus, when compared with normal renal tissue. the nuclear steroid uptake by human renal carcinomas was low for aldosterone, increased for testosterone and variably increased for progesterone. This suggests that some human renal tumours may be susceptible for steroidal regulation.
177. Effects of diethylstilbestrol
upon the morphology function of mouse testes in relation to Leydig tumorigenesis
and cell
HUSEHY. R. A. and MC‘CORMA~K. S. A., American Medical Center at Denver, Denver. Colorado. U.S.A. Continuous treatment of mice of certain strains with large doses of estrogens results in the development of Leydig cell tumors, while it is ineffective in producing such neoplasms in other strains. In two “susceptible” strains and in five F, hybrid crosses, “large dose” estrogen therapy failed to suppress spermatogenesis very significantly while in animals of two resistant strains. as well as the hybrids between them, such treatment greatly decreased testicular size and severely inhibited spermatogenesis. Testosterone (T) levels were evaluated by radioimmunoassay employing sera from individual mice of one high (BALB/c) and one low (C,H) tumor strain as well as from a F, hybrid cross of intermediate susceptibility. The data indicate that there is a tremendous variation in serum T between samples from mice of the same strain under controlled conditions of housing and bleeding. Despite this, BALB/c mice had a significantly higher level of serum T than did C,H mice. T levels in the
“intermediate” hybrid crash were c\en louer than thohc 111 the non-susceptible animals. In all genetlc type\. DES suppressed T strum levels. In treated BALB:c anomaly the T levels were as high as those 1n the untreated (‘,H mice
178. Specific binding of testosterone (T) and Sr-dihydrotestosterone (5~DHT) to plasma proteins in human males with benign prostatic hypertrophy, prostatic carcinoma, after hypophysectomy and after orchidectomy HORSI-. H.-J.. BFC~~I~R.H. and VOIC,I, K. D.. Internal
Medicine, Hamburg.
Dept. of Clinical Chcmihtr>. Hamburg. West German>
l.Jni\,cr\it)
01
Since both prostatic carcinoma (pr. ca.) and benign prostatic hypertrophy tbph) seem to be dependent on alter:)tions in secretion. and/or metabolism of androgcn,. the binding of the hormones could also be changed in these diseases. The problem therefore was inlehtigated by mcanh of a new technique for the mcasurcment of the specificall!, bound testosterone (T) and ~7-dihydrotc~toater~llie (5xDHT). Concentrations in plasma of T and 5x-DHT were determined using a radioimmunoassay and a competitive binding protein (CPB) technique respectively. In normal males an age dependent increase of specitically bound T from 30 to 5Sng,‘lOOml plasma occurred. the percentage bound T increasing from 4.3 to X.5. Sr-DHT was hound in approximately the same absolute amounts: the pcrcentage bound hormone. however. ranged from 55 to ho”,,. Despite normal blood values of T and 5x-DHT in bph. the bound fractions were 1 and I.3 times higher than occurring normally. In pr.ca. the pel- cent binding \vah lower than in bph: Estrogen treated patients with pr.ca. showed hirhlv increased value5 for both T and 5x-DHT binding. T-concentration was low and ir-DHT concentrution clearly elevated in some of these patients. Hypophysectomy concomitant with withdrawal of estrogcnh resulted in a sharp decrease of the elevated binding values found during estrogen treatment. whereas orchidectom) had an opposite effect. In one case the rfl‘ect of a combined infusion of LH and T on the snecific binding of T and Sr-DHT has been studied. Wlth’increas!ng an;ount\ of T (and LH) the specific binding of T and 5x-DHT dccrea\cd (Supported by the DFG. Sonderfor\chungsbereich 3-l “Endocrinology”~.
179. Studies on the distribution
of H.‘-17/j-oestradiol
and
H3-progesterone in normal and malignant tissues of the rat using a new autoradiographic technique
TRAMS, G. and HOLSTEIN. A.-F.. Department 01 Obstetricsand Gynaecology, Department of Anatomy. University of Hamburg. Germany The cellular and subcellular distribution of H.‘-I 7/ioestradiol and H’-progesterone was studied by autoradiographyin different organsof the female rat. For this purpose a special semi-thin-section technique wab developed which satisfied the criteria which have to he fultillcd working on the localization of steroid hormones. After injection of H”-oestradiol most radioactivity was obbrrved within the nuclei of the uterus cells. Pre-treatment of the animals with unlabelled progesterone had no effect on the uptake of subcellular location of oestradiol. This give> some evidence that progesterone does not influcncc the transport of the oestrddiol-receptor complex into the nucleus of the
Abstracts
tation. equilibrium dissociation constants of the hormonecomplexes were determined in the clone M1, : K, = 0.49 + 0.04 nM i’or the androstandlone binding to the A-receptor. and 0.41 f 0.04nM for estradiol binding to the E-receptor. The concentration of the hormone receptors in the cytosol of the clone cells were 79 & 0.04 pM/mg protein for the A-receptor and 49 i 0.02 pMimg protein for the E-receptor. The MI, clone cells contained approx. 20 times more A-receptor and approx. IO times more E-receptor than the primary culture cells.
176. Steroid hormone receptors in human renal carcinoma FANESTIL, D. D., VAUGHN. D. A. and LUDENS. J. H.. Department of Medicine, University of California. San Diego. La Jolla, California. U.S.A. Slices (0.5 mm thick) from 5 human renal adenocarcinomas and from adjacent normal renal tissue were incubated in Krebs-Ringer solution for 60 m at 37’ C in the presence of 6 x 10m9 M H”-aldosterone. H3-testosterone or H3-progesterone (in 4 cases) both in the absence and in the presence of 6 x IO-’ M non-radioactive steroid of the respective type. Cytosol (105.000 pg supernatant) and purified nuclei (PN) obtained by sedimenting the 6OOpg pellet through 2.2 M sucrose were prepared from ail slices. The CH,Cl,-extractable H3-steroid bound to protein was determined in cytosol after Bio-gel filtration and in PN. The amount of HA-steroid displaced by the lOO-fold excess of non-radioactive steroid was taken as “specific binding”. In general, the tumour cytosol contained Z-3 times more specifically bound steroid than normal cytosol. However, the nuclear specific binding was more variable. The normal and tumour nuclei contained 3.7 + 3.5 (mean i SEM) and 0.5 + 0.3 x IO ” mot aldosterone/mg protein; for testosterone the respective values were 1.05 i 3.9 and 15.7 k 8.1; for progesterone I.70 + .86 and 18.7 + 17.6. Thus, when compared with normal renal tissue. the nuclear steroid uptake by human renal carcinomas was low for aldosterone, increased for testosterone and variably increased for progesterone. This suggests that some human renal tumours may be susceptible for steroidal regulation.
177. Effects of diethylstilbestrol
upon the morphology function of mouse testes in relation to Leydig tumorigenesis
and cell
HUSEHY. R. A. and MC‘CORMA~K. S. A., American Medical Center at Denver, Denver. Colorado. U.S.A. Continuous treatment of mice of certain strains with large doses of estrogens results in the development of Leydig cell tumors, while it is ineffective in producing such neoplasms in other strains. In two “susceptible” strains and in five F, hybrid crosses, “large dose” estrogen therapy failed to suppress spermatogenesis very significantly while in animals of two resistant strains. as well as the hybrids between them, such treatment greatly decreased testicular size and severely inhibited spermatogenesis. Testosterone (T) levels were evaluated by radioimmunoassay employing sera from individual mice of one high (BALB/c) and one low (C,H) tumor strain as well as from a F, hybrid cross of intermediate susceptibility. The data indicate that there is a tremendous variation in serum T between samples from mice of the same strain under controlled conditions of housing and bleeding. Despite this, BALB/c mice had a significantly higher level of serum T than did C,H mice. T levels in the
“intermediate” hybrid crash were c\en louer than thohc 111 the non-susceptible animals. In all genetlc type\. DES suppressed T strum levels. In treated BALB:c anomaly the T levels were as high as those 1n the untreated (‘,H mice
178. Specific binding of testosterone (T) and Sr-dihydrotestosterone (5~DHT) to plasma proteins in human males with benign prostatic hypertrophy, prostatic carcinoma, after hypophysectomy and after orchidectomy HORSI-. H.-J.. BFC~~I~R.H. and VOIC,I, K. D.. Internal
Medicine, Hamburg.
Dept. of Clinical Chcmihtr>. Hamburg. West German>
l.Jni\,cr\it)
01
Since both prostatic carcinoma (pr. ca.) and benign prostatic hypertrophy tbph) seem to be dependent on alter:)tions in secretion. and/or metabolism of androgcn,. the binding of the hormones could also be changed in these diseases. The problem therefore was inlehtigated by mcanh of a new technique for the mcasurcment of the specificall!, bound testosterone (T) and ~7-dihydrotc~toater~llie (5xDHT). Concentrations in plasma of T and 5x-DHT were determined using a radioimmunoassay and a competitive binding protein (CPB) technique respectively. In normal males an age dependent increase of specitically bound T from 30 to 5Sng,‘lOOml plasma occurred. the percentage bound T increasing from 4.3 to X.5. Sr-DHT was hound in approximately the same absolute amounts: the pcrcentage bound hormone. however. ranged from 55 to ho”,,. Despite normal blood values of T and 5x-DHT in bph. the bound fractions were 1 and I.3 times higher than occurring normally. In pr.ca. the pel- cent binding \vah lower than in bph: Estrogen treated patients with pr.ca. showed hirhlv increased value5 for both T and 5x-DHT binding. T-concentration was low and ir-DHT concentrution clearly elevated in some of these patients. Hypophysectomy concomitant with withdrawal of estrogcnh resulted in a sharp decrease of the elevated binding values found during estrogen treatment. whereas orchidectom) had an opposite effect. In one case the rfl‘ect of a combined infusion of LH and T on the snecific binding of T and Sr-DHT has been studied. Wlth’increas!ng an;ount\ of T (and LH) the specific binding of T and 5x-DHT dccrea\cd (Supported by the DFG. Sonderfor\chungsbereich 3-l “Endocrinology”~.
179. Studies on the distribution
of H.‘-17/j-oestradiol
and
H3-progesterone in normal and malignant tissues of the rat using a new autoradiographic technique
TRAMS, G. and HOLSTEIN. A.-F.. Department 01 Obstetricsand Gynaecology, Department of Anatomy. University of Hamburg. Germany The cellular and subcellular distribution of H.‘-I 7/ioestradiol and H’-progesterone was studied by autoradiographyin different organsof the female rat. For this purpose a special semi-thin-section technique wab developed which satisfied the criteria which have to he fultillcd working on the localization of steroid hormones. After injection of H”-oestradiol most radioactivity was obbrrved within the nuclei of the uterus cells. Pre-treatment of the animals with unlabelled progesterone had no effect on the uptake of subcellular location of oestradiol. This give> some evidence that progesterone does not influcncc the transport of the oestrddiol-receptor complex into the nucleus of the