Sex steroid receptor proteins in foetal, adult and malignant human liver tissue.

Sex steroid receptor proteins were studied in human normal liver and hepatocellular carcinoma (HCC). Oestrogen receptor (ER) was detected in nucleosol and cytosol of 4 normal adult and 5 malignant liver specimens and in the cytosol of 6 foetal liver samples. Levels were 27.6-500 fmol mg-1 soluble protein in normal adults (Kd 1.48 X 10(-8) -1.12 X 10(-10) mol 1(-1) ), 45-290 fmol mg-1 in malignant liver tissue (Kd 3.26 X 10(-9) -3.64 X 10(-10) mol 1(-1] and a mean of 93 fmol mg-1 in foetal tissue (Kd 1.55 X 10(-9) mol 1(-1]. Androgen receptors (AR) were found only in cytosol and nucleosol of HCC (23-370 fmol mg-1) and in cytosol from foetal liver (29 fmol mg-1) with Kd from 2.90 X 10(-9) to 3.734 X 10(-10) mol 1(-1]. AR was distinguished from sex hormone binding globulin, which was also present in all cytosol samples, by the former''s ability to selectively bind to methyltrienolone and the latter''s absence from nucleosol. These findings provide further support for suggestions that oestrogen-related hepatic functions in man may be mediated by receptors and raise the possibility that hepatocellular carcinoma may be androgen dependent.     

Differences in oestrogen receptors in malignant and normal breast tissue as identified by the binding of a new synthetic progestogen

Oestrogen receptor protein (ER) was detected in 9 of 11 samples of malignant breast tissue and 8 of 9 samples of normal breast tissue. Levels of cytosolic ER (ERc) in malignant breast were 21-1102 fmol mg-1 soluble protein (Kd 1.8 X 10(-9)-3.1 X 10(-8) mol l-1) and those of nucleosolic ER (ERn), 13-526 fmol mg-1 soluble protein (Kd 2.1 X 10(-9)-1.4 X 10(-8) mol l-1). In normal breast tissue ERc levels were 33-640 fmol mg-1 soluble protein (Kd 1.3 X 10(-10)-3.2 X 10(-9) mol l-1), ERn was detected in only 2 samples, 8 and 87 fmol mg-1 soluble protein with Kd 3.2 X 10(-9) and 1.4 X 10(-9) l mol-1 respectively. 17 alpha-ethinyl-13 beta-ethyl-17 beta-hydroxy-4,15-gonadiene-3-one (gestodene), a new synthetic progestogen displaced 3H-oestradiol (3H-E2) from both ERc and ERn in malignant tissue but not in normal breast, or these receptors from endometrial tissue. In competition studies gestodene was approximately 3 times more effective in displacing 3H-E2 from ERc and ERn in malignant breast tissue than the natural ligand. Quantitation of ER by gestodene were ERc, 12-1134 fmol gestodene bound mg-1 soluble protein (Kd 1 X 10(-9)-8.1 X 10(-9) mol l-1); ERn, 17-531 fmol gestodene bound mg-1 soluble protein (Kd 1.6 X 10(-9)-1.1 X 10(-8) mol l-1). L-13-ethyl-17 alpha-ethinyl, 17 beta-hydroxy-gonen-3-one (levonorgestrel) showed no binding to ER in malignant breast, normal breast or endometrial tissue. In circulation both gestodene and levonorgestrel displaced E2 from sex hormone binding globulin more than any of the androgens tested. These results suggest that gestodene is a progestogen with oestrogenic and/or antioestrogenic properties and provide strong evidence for differences in ER from malignant and normal breast tissue.     

Presence of steroid receptors in human soft tissue sarcomas of diverse histological origin

The incidence of cytosol receptors for androgen, estrogen, glucocorticoid, and progesterone was determined in a series of histologically diverse soft tissue sarcomas. Receptor binding was characterized in tumor specimens from 29 adult patients (14 female, 15 male) obtained during surgery. Nineteen (66%) tumor cytosols bound at least one steroid. Seven (24%) had more than one receptor. Ten of 23 (43%) cytosols assayed for androgen binding were positive (Kd 0.97 +/- 0.23 X 10(-9) M; 60.6 +/- 25.9 fmol/mg protein), 7 of 29 (24%) for estrogen (Kd 1.00 +/- 0.19 X 10(-9) M; 33.2 +/- 10.9 fmol/mg protein), 9 of 26 (35%) bound glucocorticoid (Kd 4.22 +/- 1.77 X 10(-9) M; 231.0 +/- 155.0 fmol/mg protein), and 1 of 28 (4%) bound progesterone (Kd 0.41 X 10(-9) M; 18.7 fmol/mg protein). Density gradient analysis suggested that androgen and estrogen binding was located predominantly in the 6S and 7 to 8S regions, respectively, whereas receptor for glucocorticoid sedimented at 8S. Steroid binding was not related to patient age. Estrogen-positive (71%) and glucocorticoid-positive (78%) cytosols appeared predominantly in tumors from female patients. Receptor distribution also appeared to be correlated to tumor histogenetic origin.     

Estrogen receptors in hepatocellular carcinoma

Estrogen receptors (ER) were assayed on hepatocellular carcinoma (HCC) and surrounding liver tissue in 30 adult patients. All specimens were obtained at the time of surgery. Cirrhosis of the liver was associated with 28 patients and chronic hepatitis in 2 patients. ERs were detected in 12 of 30 HCCs. The value ranged from 1.4 to 9.2 fmol/mg cytosol protein with the dissociation constant (Kd) value less than 1 nanomol. On the other hand, 13 of 28 cirrhotic livers had measurable amounts of the receptors that ranged from 1.5 to 4.1 fmol/mg cytosol protein. Two livers with chronic hepatitis did not have detectable amounts of ERs. The receptors were not detected in both the tumor and liver in ten patients. The ER titers in HCC did not have any correlation with serum levels of alpha-fetoprotein or carcinoembryonic antigen, hepatitis B virus profiles, and histologic types of the tumor. In the light of the current results, it would be of great interest whether hormone therapy can be used or not as a treatment of naturally occurring HCC in humans.     

人胃癌NKM—45细胞株胞浆,胞核性激素受体含量测定及分析

作者采用葡聚糖包被活性炭法测定了人胃低分化腺癌ＮＫＭ－４５培养细胞中分离的胞浆及胞核ＫＣＩ抽提组分中的ＥＲ、ＰＲ和ＡＲ的含量。结果显示胃癌ＮＫＭ－４５细胞的胞浆及胞核抽提液中ＥＲ含量为２４．８ｆｍｏｌ／ｍｇ和２５．１ｆｍｏｌ／ｍｇ蛋白，而ＰＲ含量为１６．４ｆｍｏｌ／ｍｇ和１８．６ｆｍｏｌ／ｍｇ蛋白，ＡＲ均为阴性（＜１０ｆｍｏｌ／ｍｇ蛋白），说明ＮＫＭ－４５细胞为ＥＲ及ＰＲ阳性细胞，且ＥＲ和ＰＲ在该胃癌细胞的胞浆、胞核中均存在。作者认为胃癌可能是性激素依赖性肿瘤，适当的内分泌治疗可能对胃癌有效。     

Sex hormone receptors in gastric cancer

Gastric adenocarcinoma that originates from mucosal tissue invades submucosa, muscle, and serosa in different stages. The level of progesterone receptors (PgR), estrogen receptors (ER), and androgen receptors (AdR) in the superficial part of gastric cancer tissues (CAs) from 16 patients was determined and compared with that of the corresponding normal gastric mucosal tissues (NLm). There were PgR in all CAs (100%) with values that ranged from 20.5 to 548.4 fmol/mg protein. Eight CAs (50%) had ER values that ranged from 6.8 to 325.1 fmol/mg protein. AdR was found in two CAs with values of 14.7 and 16.4 fmol/mg protein. In NLm, 15 (93.8%) had PgR values that ranged from 7.3 to 473.2 fmol/mg protein and ten (62.5%) had ER values that ranged from 0.9 to 87.9 fmol/mg protein. AdR were present in two NLm with values of 1.5 and 73.5 fmol/mg protein. There was no statistical difference in levels of PgR and ER between CAs and NLm. There were PgR in all gastric cancers and in 93.8% of NLm. The results suggest that gastric mucosa may be the target tissues for progesterone action. Furthermore, the lack of correlation between the levels of ER and PgR in gastric cancer tissue suggests that the PgR in gastric cancers are probably estrogen independent.     

Hormone receptor status in human endometrial adenocarcinoma

Steroid receptor levels were determined in 196 samples of endometrial adenocarcinoma: cytosol estradiol receptors (ERc) were measured in 171 samples, cytosol progesterone receptors (PRc) in all samples; nuclear estradiol receptors (ERn) and nuclear progesterone receptors (PRn) in 68 samples; total estradiol receptors (ERt = ERc plus ERn) and total progesterone receptors (PRt = PRc plus PRn) were measured in 68 samples. The ERc levels were 88.2 +/- 8.9 (mean +/- SEM) and ERn were 94.4 +/- 15.6 fmol/mg protein; PRc levels were 197.9 +/- 25.9 and PRn 178.3 +/- 55.9 fmol/mg protein. The ERt levels were 162.6 +/- 23.2 and PRt 249.8 +/- 75.7 fmol/mg protein. The presence of PRc was related to the ERc levels according to the cut-off used. Estradiol receptors (ER) and progesterone receptors (PR) were present in the cytoplasmic and nuclear fractions in 60.2% and 36.8% of cases, respectively. The simultaneous presence of both ERt and PRt was observed only in 27.9% of cases. In the normal endometrium ERc and PRc were negatively correlated (r = -0.525, P less than 0.005), whereas in endometrial adenocarcinoma the correlation was positive (r = 0.491, P less than 0.001). In contrast with the normal endometrium the correlation between ERc and ERn was positive (r = 0.582, P less than 0.001) in tumor tissue. In neoplastic tissue Scatchard analysis showed a single class of specific ERc sites with a dissociation constant (Kd) of 1.39 +/- 0.8 X 10(-9) mol/l, one tenth of that found in the normal premenopausal endometrium. Qualitative and quantitative analysis of the receptor status showed that in 30% to 40% of cases studied the behavior of the neoplastic cell was similar to that found in the normal endometrial cell. In a 4-year follow-up of patients affected by endometrial adenocarcinoma there is better survival in the groups of patients with a simultaneous presence of ERt and PRt than in the group with their absence.     

Acetylsalicylic acid (ASA) protects the prostaglandin-cAMP-system of human hypernephroma cells against irradiation-induced alterations.

There is abundant evidence that inhibitors of prostaglandin (PG) biosynthesis might increase the radioresponse of certain tumour cells. This study investigated specific PG binding sites, eicosanoid production as well as intracellular cAMP levels in cultured human hypernephroma cells derived from 11 patients upon nephrectomy. Scatchard analyses of the binding data revealed specific PGE1-, PGE2- as well as PGI2-binding sites (PGE1: Bmax = 755 +/- 206 fmol mg-1 protein, Kd = 3.7 +/- 2.7 nM PGE2: Bmax = 494 +/- 221 fmol mg-1 protein, Kd = 4.2 +/- 2.5 nM; PGI2: Bmax = 693 +/- 164 fmol mg-1 protein, Kd = 6.0 +/- 4.5 nM). Significant (P < 0.01) increase in PG binding sites expressed on human hypernephroma cells (PGE1: Bmax = 1084 +/- 303 fmol mg-1 protein, Kd = 2.8 +/- 1.3 nM; PGE2: Bmax = 663 +/- 309 fmol mg-1 protein, Kd = 2.2 +/- 1.5 nM; PGI2: Bmax = 1021 +/- 391 fmol/protein, Kd = 4.2 +/- 3.6 nM) and inhibition of PG biosynthesis (TXB2: -82.5%, PGE2: -87.5%. PGD2: -80.6%, PGF2: -81.3%) were found after acetylsalicylic acid (ASA)-treatment (0.5 mg 10(-6) cells for 24 h). Following irradiation (60Co, 1.0 Gy/min-1 over 10(min), PG binding sites (PGE1: Bmax = 266 +/- 153 fmol mg-1 protein, Kd = 5.0 +/- 5.0 nM; PGE2: Bmax = 148 +/- 66 fmol mg-1 protein, Kd = 4.7 +/- 3.6 nM; PGI2: Bmax = 325 +/- 194 fmol mg-1 protein, Kd = 6.8 +/- 7.1 nM) were significantly (P < 0.01) diminished. However, irradiation had no significant effect on PG binding sites in ASA-pretreated cells (PGE1: Bmax = 699 +/- 240 fmol mg-1 protein, Kd = 3.5 +/- 1.8 nM; iloprost: Bmax = 766 +/- 452 fmol mg-1 protein, Kd = 3.2 +/- 2.2 nM). Although there was no significant difference in the basal values for cAMP between control and ASA-treated group cells, the PG-induced cAMP-production was less pronounced in the control group. Taken together, the findings suggest that ASA may modify the radioresponse of cultured human hypernephroma cells by preventing the decrease of PG binding sites induced by irradiation.     

Nuclear translocation of the estrogen receptor in autonomous C3H mouse mammary tumors

The reason for estrogen independence of C3H mouse mammary tumors has been sought in the initial steps of estradiol action. The characteristics of the estrogen receptors were similar to those observed in estrogen-responsive tissues: high affinity and binding specificity, DNA binding and 8S sedimentation constant as shown by sucrose gradient centrifugation. Their concentration averaged 18.5 +/- 3.5 (S.E.) fmol/mg cytosol protein in the cytosol and 3.5 +/- 1.0 fmol/mg cytosol protein in the KCl nuclear extract. The nuclear translocation of the cytosol receptor was investigated with the use of biopsy and in vivo injections of radioactive estradiol. No nuclear translocation of estrogen receptor could be ascertained with the dextran-coated charcoal assay since the free and nonspecifically bound estrogen conjugate(s) were also assayed by this technique. However, when the estrogen-receptor complexes were estimated by more specific methods such as protamine sulfate or hydroxylapatite precipitations, the estrogen receptor translocation into the nucleus was clearly shown. We therefore conclude that the estrogen independence of C3H mammary tumors cannot be explained by a defect in the two initial steps of the mechanism of action of estradiol, namely, cytosol binding and nuclear translocation of receptors.     

Decrease of prostaglandin I2 binding sites in thyroid cancer

The properties of specific prostaglandin I2 (prostacyclin, PGI2) binding sites in normal thyroid tissue have been characterised. Tissue samples obtained intraoperatively from patients with 'cold' solitary thyroid nodules (as preoperatively selected by thyroid gland scintigraphy, thyroid gland ultrasonography and Papanicolaou cytology following fine needle aspiration of the nodule area) have been used for thyroid membrane preparation. Employing [3H]iloprost, a chemically stable PGI2-analogue as a radioligand, saturation experiments for comparative binding studies have been attempted. Scatchard analysis of the binding data obtained for normal thyroid parenchyma distant to the nodule area revealed heterogeneity of the [3H]iloprost sites exhibiting a high-affinity binding capacity (Bmax) of 613.2 +/- 130.4 fmol mg-1 membrane protein and a low-affinity binding capacity of 5.1 +/- 1.6 pmol mg-1 membrane protein. The equilibrium dissociation constant (Kd) amounted to 18.9 +/- 8.9 nM and to 131.5 +/- 39.2 nM, respectively. Scatchard analysis of the binding data obtained for benign thyroid adenoma indicated significant lower binding capacities exhibiting a Bmax of 325.8 +/- 110.0 fmol mg-1 membrane protein (Kd: 31.0 +/- 7.5 nM) for the high-affinity sites and of 3.9 +/- 2.5 pmol mg-1 membrane protein (Kd: 364.9 +/- 183.6) for the low affinity sites. In cancer tissue a selective loss of the low affinity sites and a significant diminution of the high-affinity sites was observed: in well differentiated cancer the high-affinity sites showed a Bmax of 299.7 +/- 46.0 fmol mg-1 membrane protein (Kd: 38.9 +/- 7.3 nM), in anaplastic cancer, less differentiated papillar and follicular cancers of 180.6 +/- 25.1 fmol mg-1 membrane protein (Kd: 54.6 +/- 16.7 nM). Well differentiated papillar and follicular cancers did not differ from each other.(ABSTRACT TRUNCATED AT 250 WORDS)     

雌二醇对胃癌细胞周期的影响

目的:初步探讨雌二醇（E2）对胃癌细胞SGC-7901增殖的影响。方法:SGC-7901细胞分为E2干预组和对照组,E2干预组加入不同浓度E2（0.1μmol/L、1.0μmol/L）干预24小时,对照组加入等体积含无水乙醇的培养基。CCK8检测E2对胃癌细胞增殖能力的影响,流式细胞术检测细胞周期的分布,PCR检测E2对胃癌细胞雌激素受体ERα和ERβ mRNA表达水平的影响,Oncomine公共数据库查询ERα的表达情况。结果:E2对胃癌细胞的增殖抑制率具有浓度依赖性。E2处理组SGC-7901细胞G0/G1期细胞比例［(68.866±3.336)%］较对照［(59.333±0.294)%］显著增加,G2期和S期细胞总比例下降,1.0μmol/L E2作用可使胃癌细胞SGC-7901发生G1期阻滞（P<0.01）。两种雌激素受体（ERα和ERβ）均表达于胃癌细胞SGC-7901,且其相关基因ERα和ERβ mRNA表达水平不随E2浓度的增加而改变。利用Oncomine公共数据库查询发现ERα在胃癌中的表达高于癌旁。结论:E2直接作用可抑制胃癌细胞SGC-7901的增殖,引起G1期阻滞。     

Status of sex steroid hormone receptors in large bowel cancer

To determine the potential role of sex steroid hormones in the development of colorectal tumors in humans, specific androgen (AR), estrogen (ER), and progesterone (PGR) receptors were investigated in normal mucosa (NM) and in tumor (T) paired biopsy specimens from 94 patients. Androgen receptors were detected in 98% and 96% of NM and T samples, ER in 91% and 83% of NM and T biopsy samples, whereas PGR were detected only in 14% and 10% of NM and T specimens, respectively. These incidences are independent of the sex and age of the patients. They are not related to tumor localization, histologic grade, or stages of Dukes' classification. Scatchard analysis of labeled ligand binding indicated the existence of one single class of high affinity binding sites; the calculated dissociation constant (Kd) was 1.7 +/- 0.6 10(-9) molar concentration (M) for 5 alpha-dihydrotestosterone (DHT) and 0.6 +/- 0.3 10(-9) M for estradiol (E2). These values were identical in NM and T tissue for both AR and ER. The binding capacity for DHT was 148 +/- 67 and 93 +/- 43 fmol/mg of cytosol protein in NM and T tissues, respectively (P less than 0.05). The ER content was lower and similar in NM and T biopsy specimens: 19 +/- 9 and 18 +/- 10 fmol/mg protein, respectively. The PGR content was 10 +/- 4 in NM versus 17.5 +/- 6 fmol/mg protein in T specimens. It is observed that the elevated AR in normal mucosa is not related to any known function for androgens in the digestive tract. The receptor pattern observed in tumors does not support the hypothesis previously raised in the case of chemically induced colonic tumors in rodents.     

Effects of glucocorticoids on the growth of human fibrosarcoma cell line HT-1080

The human fibrosarcoma cell line HT-1080 exhibits rapid growth following s.c. inoculation in 4-6-week-old male athymic mice. Cytosols from tumors carried in athymic mice bind glucocorticoid (Kd, 1.8 +/- 0.48 X 10(-8) M; Bmax, 240.5 +/- 35.3 fmol/mg cytosol protein, mean +/- SEM). Receptor sediments primarily in the 8-9S region on 5-20% sucrose gradients and is specific for the glucocorticoids. HT-1080 growth in vitro (as measured by cell count) was inhibited over a range of 10(-6)-10(-8) M after 7 days of incubation with dexamethasone and triamcinolone acetonide. Progesterone, estradiol, and dihydrotestosterone had no effect on HT-1080 growth in vitro. Preincubation with a 100-fold excess of progesterone reversed the growth inhibition observed with triamcinolone acetonide but not dexamethasone acetate. HT-1080 tumor cell growth responded biphasically to dexamethasone in vivo. Athymic mice given s.c. injections every other day with 5 or 25 micrograms dexamethasone showed an increase in tumor size inversely proportional to dose. In contrast, 200 micrograms of dexamethasone significantly inhibited tumor growth. Adrenalectomy did not significantly alter HT-1080 growth or glucocorticoid binding to tumor cytosols (Kd, 3.4 X 10(-8) +/- 1.1, Bmax, 236.9 +/- 9.9 fmol/mg cytosol protein, mean +/- SEM) although tumor incidence was decreased in sham adrenalectomized mice. Glucocorticoid binding in tumors grown in vivo was decreased by increasing amounts of dexamethasone. High pharmacological doses of glucocorticoids inhibit the growth of human fibrosarcomas in vivo and in vitro.     

Somatostatin binding in normal and malignant human gastrointestinal mucosa.

Somatostatin is a regulatory peptide implicated in the control of cellular proliferation in epithelial tissues and this regulation may occur directly via membrane bound receptor activation. The aim of this study was to investigate somatostatin binding in human gastrointestinal cancer and normal mucosa. Plasma membranes were prepared from specimens of tumour and normal mucosa from 51 patients undergoing surgical resection for malignancy (28 gastric, 23 colorectal). Using a competitive displacement assay, specific 125I-tyrosine-11-somatostatin-14 binding to plasma membranes was assessed and and characterised in terms of receptor affinity (Kd) and maximum binding capacity (Bmax) as determined by Scatchard analysis. Specific low affinity (Kd = 166 nM), high capacity (Bmax = 1.2 pmol mg-1 protein) somatostatin binding was demonstrated in 22 of the gastric cancers and 17 of the colorectal cancers (Kd = 140 nM, Bmax = 1.8 pmol mg-1 protein). Similar affinity and binding capacity was demonstrable in normal mucosal samples. High affinity receptors for somatostatin were expressed by one gastric carcinoma (Kd = 0.9 nM; Bmax = 0.23 pmol mg-1 protein). Thus, low affinity, high capacity binding is a common feature of gastrointestinal tumours and normal mucosa, and high affinity receptors may occasionally be demonstrated. The functional significance of these low affinity binding sites requires elucidation to determine whether long-acting somatostatin analogues may have therapeutic benefit in gastrointestinal malignancy.     

Comparative studies of estrogen receptor determinations by enzyme immuno-assay using the monoclonal antibody, dextran-coated charcoal, and sucrose density gradient methods

Estrogen receptor (ER) determination in human mammary tumors was performed by enzyme immunoassay (EIA) using monoclonal antibody, and the results were compared with those obtained using the dextran-coated charcoal (DCC) and sucrose density gradient (SDG) method. Twenty ER-positive tumors by the DCC method were all ER-positive by EIA, when determined in the same cytosol fraction. Three out of 21 ER-negative tumors by the DCC method were ER-positive by EIA. Estrogen receptors of 8S, 8S + 4S, and 4S determined by the SDG method were all shown to be ER-positive using EIA. Between the values of estrogen receptors determined by the DCC method and by EIA, a high correlation was observed (r = 0.868). Values of ER less than 13.5 fmol/mg protein were considered as negative ER.     

Comparison of steroid receptor levels in renal-cell carcinoma and autologous normal kidney

Since a number of renal-cell carcinomas regress with hormonal manipulation, we have identified and measured the levels of estrogen, progestin and glucocorticoid receptors in 47 autologous pairs of normal and neoplastic kidney tissues. High-affinity receptors for these hormones were detected in kidney tissues of both sexes by means of a dextran-coated charcoal assay. Glucocorticoid receptors were demonstrated in renal cancer tissues for the first time, and were higher in the tumor (mean 31.3 ± sem 5.6) than in the normal tissue (mean 18.5 ± sem 3.1 fmol/mg cytosol protein). There was a significant difference in the quantities of progestin receptors (expressed as fmol/mg cytosol protein) in normal (mean 18.4 ± sem 3.3) versus neoplastic (mean 10.4 ± sem 4.0) kidney specimens (p < 0.007). There was a significant difference between the binding affinity of the progestin receptor in the male tumors (Kd = 2.2 ± sem 0.9 nM, n = 10) and that of the females, (Kd = 9.3 ± sem 6.5 nM) (p < 0.04). When an affinity of < 9.9 × I0^?9M and > 10 fmol/mg cytosol protein were used as criteria for classifying a tissue as positive for progestin receptors, only 17% of tumors contained these receptors while 45% of normal tissues exhibited them. According to these criteria, no differences were observed in the frequency of occurrence of either estrogen receptors or glucocorticoid receptors in tumor versus normal kidney. Data from this study suggest that the use of endocrine therapy should be re-examined in the treatment of renal-cell carcinoma.     

A study of the number of binding sites of estrogen receptors as a malignancy factor in primary breast cancer

Whether or not estrogen receptor (ER) is an indicator of malignancy in breast cancer remains to be clarified. Although there are a number of reports on the prognosis examined in relation to the presence or absence of ER, few studies have examined the number of ER binding sites. We investigated whether or not the number of ER binding sites could serve as an indicator of malignancy in ER positive patients. The subjects were 109 patients with primary breast cancer who underwent surgery between 1985 and 1987. The dextran-coated charcoal assay was used to determine ER. The number of ER binding sites over 5 fmol/mg cytosol protein (fmol/mg c.p.) was regarded as positive. The ER positive rate among all patients with breast cancer was 69.7%. Analysis of the relations of ER positive rate to each grade of the stage, n factor, menstruation, CA15-3 and nuclear DNA content revealed no correlation with any of these factors, except for the nuclear DNA content, suggesting that the presence or absence of ER alone does not indicate malignancy. There was a significant difference between the average number of ER binding sites of pre-climacteric patients and that of post-climacteric patients, 32.31 +/- 26.72 fmol/mg c.p. and 126.94 +/- 125.88 fmol/mg c.p., respectively. So it is considered that the analysis of the number of ER binding sites should be evaluated according to their menstrual states.(ABSTRACT TRUNCATED AT 250 WORDS)     

人胃癌SGC-7901细胞胞浆胞核性激素受体测定

 作者采用葡聚糖包被活性碳饱和分析法(DCC法)分别测定了培养的人胃低分化腺癌SGC-7901细胞的胞浆及胞核KCL抽提液中的雌激素受体(ER)、孕激素受体(PR)和雄激素受体(AR)的含量。 结果证明胃癌SGC-7901细胞的胞浆及胞核抽提液中ER含量为30.2和35.7fmol/mg蛋白，而PR的含量为20.3和22.7fmol/ng蛋白, 但AR均为阴性(<10fmol/mg蛋白), 这说明SGC-7901细胞为ER及PR阳性细胞, 其ER和PR在胞浆、胞核中均有分布。 提示胃癌可能为性激素依赖性肿瘤, 有内分泌治疗的可能。     

Correlation of lactogenic receptor concentration in human breast cancer with estrogen receptor concentration

The presence of receptors for lactogenic hormones in human breast cancer tissue has been documented previously, but the relationship between the expression of these receptors and estrogen receptor (ER) status has not been adequately studied. In this report, the specificity of 125I-human growth hormone (HGH) binding in both cultured human breast cancer cell lines and tumor biopsies was studied to establish that HGH was a suitable ligand for investigating lactogenic receptor concentration in these tissues. In addition, the relationship between specific binding of 125I-HGH and ER concentration in human breast cancer was investigated. Specific 125I-HGH binding to 14 breast cancer cell lines in long term culture and to membrane preparations (microsomal and plasma membrane fractions) from 31 breast cancer biopsy specimens was examined. Human prolactin and HGH were approximately equipotent in inhibiting binding of 125I-HGH to both cultured breast cancer cell lines and to membrane preparations from breast cancer biopsy specimens. Competitive inhibition experiments using lactogenic and somatogenic hormones established that the specificity of 125I-HGH binding to breast cancer biopsy material was similar to that of cultured breast cancer cell lines and similar to that reported for subprimate lactogenic receptors. Saturable, high-affinity (Ka = 0.53 to 2.33 nM-1), low-capacity (330 to 6560 sites/cell) growth hormone binding sites were found on each of the ER-positive cell lines, whereas no specific 125I-HGH binding to ER-negative cell monolayers was detected. When all cell lines were considered, a significant linear correlation (r = 0.745, p less than 0.001) between ER and lactogenic receptor concentrations was found. Significant specific 125I-HGH binding, greater than 1% of the total radioactivity added, was detected in 20 of 31 (65%) breast tumor biopsy specimens. The mean affinity and capacity of the lactogenic receptor as measured in 8 separate membrane preparations were Ka = 0.52 +/- 0.09 (S.E.) nM-1 and 255 +/- 85 fmol/mg protein. Membrane preparations from ER-negative tumors (less than 3 fmol ER/mg cytosol protein) bound significantly less 125I-HGH than did membrane preparations from ER-positive tumor biopsies (1.22 +/- 0.44 versus 3.21 +/- 0.56%, p less than 0.05). A significant linear correlation between specifically bound 125I-HGH and ER concentration (r = 0.412, p less than 0.02) was demonstrated in the 31 breast cancer biopsy specimens studied.(ABSTRACT TRUNCATED AT 400 WORDS)     

Receptor characteristics of the rat mammary carcinoma cell line 64-24

Saturable binding of androgens, glucocorticoids, and triiodothyronine was found in the 64-24 hormone-responsive rat mammary carcinoma cell line. Androgen receptors had a dissociation content (Kd) for methyltrienolone of 3.4 X 10(-10) M and a binding capacity of approximately 10,000 sites/cell in whole cells. 5 alpha-[3H]dihydrotestosterone (DHT) was specifically taken up into approximately 2,150 nuclear sites with an affinity of 8.3 X 10(-10) M when nuclei were isolated from whole cells incubated with [3H]DHT. Sucrose gradient centrifugation of cytosol prepared from these cells revealed a displaceable [3H]DHT-binding component which migrated at 8S. Sedimentation analysis with high salt gradients of nuclear extracts from cells incubated with [3H]DHT revealed a peak of radioactivity in the 4S region which was abolished by coincubation of the cells with excess nonradioactive methyltrienolone. Receptors for [3H]dexamethasone were more abundant (approximately 50,000 sites/cell) in whole cells and had a Kd of 7.5 X 10(-9) M, but the number of nuclear binding sites was similar to that for androgens. Specificity studies using unlabeled steroids showed that each of the two classes of steroid receptors had greater affinities for their appropriate hormones. High affinity receptors for estrogens and progestins were not detectable in these cells. Triiodothyronine receptors were demonstrable but at a very low binding capacity (1,100 sites/cell). The Kd of these receptors was 0.6 X 10(-10) M. Cytogenetic studies revealed 44 chromosomes/mitosis with several unique markers. These receptor and karyotypic features suggest that the 64-24 cells may be useful in studying androgen action on breast cancer independently of estrogen or progestin influence, as well as the effects of thyroid hormone and glucocorticoids on breast cancer cells.