SEX HORMONE BINDING GLOBULIN BINDING
CAPACITY, TESTOSTERONE, 5\g=a\-DIHYDROTESTOSTERONE,
OESTRADIOL AND PROLACTIN IN PLASMA OF PATIENTS WITH PROSTATIC CARCINOMA UNDER
VARIOUS TYPES OF HORMONAL TREATMENT
W. Bartsch,
By H.-J. Horst, H. Becker and G. Nehse1) ABSTRACT
Sex hormone binding globulin (SHBG) binding capacity, the concentrations of testosterone (T), of 5\g=a\-dihydrotestosterone(DHT), of oestradiol\x=req-\ 17\g=b\(Oe2), of oestrone (Oe1), of prolactin (hPr) and the percentual specific binding of T to SHBG (% TB) were measured in plasma of patients suffering from prostatic carcinoma and of a control group of similar age. No significant differences in any of the investigated parameters were found between the control group and the carcinoma patients before treatment although 15 % of the latter showed distinctly elevated hPr values. Treatment of carcinoma patients with
1) Antiandrogen (cyproterone acetate, Androcur\s=r\)resulted in a signifidecrease of T, Oe2 and SHBG. The DHT/T-ratio increased. n 5. 2) Orchidectomy caused an even more pronounced fall in T, DHT, Oe1 and Oe2 blood levels. SHBG was not altered. DHT/T-ratio increased. cant
n
=
=
32.
3) Cyproterone
acetate after
orchidectomy
led to elevated hPr values.
n=5.
4) Oestrogen (diethylstilboestrol-diphosphate, Honvan\s=r\) after tomy increased SHBG and hPr. 0 Part of doctoral thesis.
n
=
6.
orchidec-
Corticosteroid (Prednisone, Decortin®) after orchidectomy decreased T and SHBG below the levels found after orchidectomy alone, n 5. 6) Diureticum (Mefruside, Baycaron®) (n 5) or 7) a placebo (n 7) did not alter any of the parameters measured. 8) Treatment with HCG (Primogonyl®) of patients suffering from oligozoospermia resulted in a significant increase of T, DHT and Oe2. SHBG 7. was not altered. DHT/T-ratio decreased, n
5)
=
=
=
=
It is well known that the prostate is an androgen dependent organ. Conse¬ quently the most common treatment of prostatic carcinoma is the withdrawal of androgens either by administration of oestrogens, anti-androgens and/or orchidectomy. These treatments reduce plasma androgen levels (Young 8c Kent 1968; Boyns et al. 1974; Jönsson et al. 1975; Harper et al. 1976) to low con¬ centrations, which then originate mainly from adrenal precursors (Cowley et al. 1976). Besides the total concentrations of T and DHT in plasma, SHBG is an important factor as it might influence the prostatic uptake of androgens. Its role in lowering the biologically available androgens at the tissue level has been clearly demonstrated by Lasnitzki 8c Franklin (1972, 1975) and Giorgi 8c Moses (1975). It has been shown that prolactin, too, has a stimulating effect on the prostate (Famsworth 1972; Lasnitzki 1972). These properties of pro¬ lactin in connection with prostatic carcinoma have been discussed by Boyns et al. (1972) and Harper et al. (1976). According to these authors a rise of prolactin release by therapy should be avoided in prostatic carcinoma. Ad¬ ditionally we found in 20 °/o of prostatic carcinoma patients elevated serum prolactin values both by radioimmunoassay and to an even higher extent by bioassay, which could be correlated with the histologically type of tumour present (Bartsch et al. 1977). Probably oestrogens also act directly on the prostate (Neumann el al. 1975; Hawkins et al. 1975; Wagner et al. 1975). Better known, however, is their inhibition of pituitary LH secretion and their potent stimulation of SHBG (Vermeulen et al. 1969) and hPr (Mortimer et al. 1974) secretion. These facts prompted us to study the plasma levels of SHBG, T, DHT, Oe2 and hPr in patients with prostatic carcinoma under the influence of various
types of
treatment.
MATERALS AND METHODS
Patients Patients with included in this
recently diagnosed and histologically proven prostatic carcinoma were study. They did not receive any therapy of known influence on endo-
crinological parameters. Conditions
of
Blood
was
drawn between 8—10
a. m.
treatment
1) Five patients received 300 mg cyproterone acetate (Androcur®) im every 2 weeks. Blood was drawn on 3 consecutive days before treatment and every 4 days after the
injections. Mean values of results before and between the injections were used for statistical analysis. 2) From all other patients blood was taken once before and 1 and 6 weeks after orchidectomy. After that time the further treatment was performed as follows: 3) Five patients received cyproterone acetate (300 mg/2 weeks im). 4) Six patients received diethylstilboestrol-diphosphate (Fosfestrol, Honvan®) (3 x 120 mg per os/d). 5) Prednisone (Decortin®) was given to 5 patients (3 x 5 mg per os/d). 6) Mefruside (Baycaron® a diureticum) was administered to 5 patients (2 x 25 mg
os/d). 7) A placebo was given to 7 patients. Treatment 3)-7) started at least 6 weeks -
per
1 week and at least 3 months after
after
orchidectomy.
Blood
was
collected
starting the therapy. 8) To evaluate opposite effects to orchidectomy patients with oligozoospermia re¬ ceived every 2nd day for 2 weeks 5000 IU HCG (Primogonyl®), total dose 30 000 IU im. Blood
was
taken before the first and 2 h after the last dose.
All treatments and sampling of blood were done with the consent of the after they were informed about the study.
patients,
Assays binding capacity was measured by the method of Dennis et al. (1977). Plasma stripped of the endogenous steroids with charcoal, SHBG precipitated with ammoniumsulfate (28 "la final concentration) and then redissolved in a buffer containing a saturating amount of DHT plus a tracer amount of radioactive DHT. A differential dissociation technique with dextran coated charcoal was used to eliminate unspecific binding. Values were recorded in Moles/1 binding capacity for DHT which should correspond to the molar amount of SHBG under the assumption of an 1:1 binding relation of SHBG and DHT. Coefficient of variation (CV) was 3.8 °/o. The porcentual amount of specifically SHBG bound T (%> TB) was estimated by the method of Horst et al. (1974) by differential dissociation in 1:5 diluted plasma after equilibrating with a tracer amount of radioactive T. Plasma T, DHT, Oe4 and Oe2 were estimated by radioimmunoassay (Bartsch et al. 1977) after extraction of 3 ml plasma with ether and separation of three steroids (DHT, Oe4, Oe2 or DHT, T, Oe2) from one sample by celite column chromatography: columns of 0.5 cm diameter were packed tightly with 0.8 g celite:ethyleneglycol:propyleneglycol 4:1:1 (w, v, v) and eluted under nitrogen pressure with 3.5 ml ¿so-octane (discarded), 3.5 ml 5 "la toluene in ¿so-octane (androstenedion, discarded), 3.0 ml 40 "la toluene in ¿so-octane (DHT), 3.5 ml 50 "la toluene in ¿so-octane (T), 3.0 ml 60 % toluene in ¿so-octane (discarded) and 3.5 ml 25 % ethylacetate in toluene (Oe2). T and Oej were not completely separated; to estimate DHT, Oe4 and Oe2 3.0 ml of the 50 "la SHBG
was
-
toluene in ¿so-octane fraction
were
discarded and 3.5 ml of the 60 "la toluene in ¿so-
containing Oe4 were collected. Appropriate amounts of the eluates were pipetted into glass vials and dried before pipetting reagents for radioimmuno¬ assay, which was performed by the dextran coated charcoal method. Antisera against T and oestrogens (100% cross reaction of Oej and Oe.j) were a generous gift of Organon Internatinal, B.V., Oss (The Netherlands). Anti-DHT antibody was pur¬ chased from Miles Corp., Slough (England). Sensitivity is 12, 20, 3, 3 pg/ml plasma for DHT, T Oe, and Oe2, respectively. The absence of systematic errors was shown by dilution experiments and addition of authentic steroids. Coefficient of variation octane
fraction
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(CV) was 10.5, 8.4, pool serum. hPr
was
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measured
by
Sprendlingen (Germany). was
a
DHT, T, Oej and Oe2 using
normal male
specific radioimmunoassay
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was
19.8 "la.
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a
kit from Isotopendienst West, NIH 1. Coefficient of variation (CV)
analysis
All results are given in geometric mean (95 "la confidence limits of the mean). Sta¬ tistical differences are evaluated by Student's paired, two tailed i-test, or comparing normals and patients by Student's multiple, two tailed i-test, after transformation of data to the logarithmic scale.
RESULTS
As summarized in Table 1 SHBG binding capacity as well as all other para¬ investigated in plasma of patients with prostatic carcinoma, were found be in the same order of magnitude as in males of similar age not suffering to clinically from prostatic diseases. 5/34 (15%) of the patients showed serum hPr higher than the 2 o limit of normal men. The elevated hPr values did not correlate with high Oe2 or SHBG levels. For comparison normal values of younger men were determined. They showed significantly lower Oe2 (P