Dextran-coated Charcoal Technique Research Articles

38P Frequent changes in the environment can lead to heterogeneity of tumour cells

The mouse mammary tumors developed on the background of changes in the hormonal environment during several cycles of estrus differ from tumors of mice developed during a stable high or stable low hormonal environment. The difference is that the changing hormonal background leads to heterogeneity in the level of estrogen and progesterone receptors in tumors. Female C3H/Sn mice were used in this study. Transplantable mammary tumors were received from spontaneous mammary tumors C3H/Sn mice. ER-positive (ER +) adenocarcinomas in 90% of cases were obtained from mice at the age of 12 months with intensive reproduction (giving birth at least 4-5 times). The level of ER was determined by means of the dextran-coated charcoal technique with the use of 3H-estradiol. The level of receptors protein in mice tumors developed with stable high level of hormones reaches up the range higher than 20,0 fmol/mg of protein up to 110 fmol/mg. The transplanted mice mammary tumors which developed against the environment of high level of estrogens (frequent parturition without mammary feeding of off-springs) contains high level of estrogen receptors. The hormonal environment of the organism determines the level of receptors in the tumors at the end of their development independent of the level of receptors in tumor before transplantation. The wavy development of tumor depending from oestrus phase of cycles during 3-5 cycles is observed in transplanted mice tumors with normal oestrus cycle (cycle changes of hormonal environment). The change of hormones level which is observed during 4-daily of mice oestrus cycle is capable to cyclic influence on change of volume under progressive growth of receptor-positive tumors. The growth loses the wavy feature after 5-th cycle. At the end of tumor development approximately half of cells lose the receptors. Thus, the population become heterogeneous in the content of estrogen receptors. Heterogenization of tumors indicates on the one hand the harmful properties of tumors to adapt to new conditions. On the other hand, the high sensitivity of tumors to changes in the environment gives hope for multi-stage therapy.

1,25-Dihydroxyvitamin D3 receptors and human colon adenocarcinoma

Epidemiological evidence suggests that dietary calcium and 1,25-dihydroxyvitamin D3 (1,25-(OH)2D3) might have a protective effect against colorectal cancers. Since the presence of receptors is required for steroid action, specific 1,25-(OH)2D3 receptors (RD3) were investigated in biopsies taken at different levels of the colon. The study involved 90 biopsies from patients operated on for colorectal adenocarcinoma. They were paired biopsies from adenocarcinoma tissue and adjacent normal mucosa. In addition, 26 normal intestinal mucosa biopsies from patients without cancer were examined. RD3 receptors were assayed in tissue extract by the dextran-coated charcoal technique and also characterized by sucrose density gradient sedimentation. Scatchard analysis showed a single class of specific high affinity-low capacity sites binding for 1,25-(OH)2D3. The incidence of RD3 was 86 per cent in normal mucosa (n = 77) and lower in carcinoma (n = 34), for which the incidence decreased significantly (P less than 0.001) from right colon (58 per cent) to left colon (37 per cent) and rectum (19 per cent). These data suggest that the normal colon is a potential target organ for 1,25-(OH)2D3 which might modulate calcium transport in the colon. Loss of receptivity to 1,25-(OH)2D3 is associated with malignant transformation.

Tissue heterogeneity of immunohistochemically detected estrogen receptor. Implications for image analysis quantification.

The increasingly popular immunohistochemical techniques for assay of the estrogen receptor (ER) allow localization of receptor positivity to specific cell populations. Heterogeneity of the ER in tumor cell populations may have important implications for analytic cell selection and for prognosis in ER-positive carcinomas. We studied 84 tissue blocks for level-to-level and geographic heterogeneity within level of the ER and cytokeratin by staining alternate serial sections for ER and cytokeratin. Distribution of ER and cytokeratin positivity was manually assessed. Homogenous positive staining was seen in 63 of 84 cases for ER and 71 of 84 cases for cytokeratin. Distinct geographic variability constant from level to level was seen in 7 cases for ER. In each of these cases, the cell populations stained uniformly for cytokeratin. Artifactual heterogeneity seemed to be uncommon for ER. Automated image analysis and manual ER estimation resulted in more positive cases than did the dextran-coated charcoal (DCC) technique. Interobserver correlation for the manual method seemed high, as did correlation between the manual method and image analysis. Because a majority of the immunohistochemical staining heterogeneity for ER seemed to be biologic, we believe it may mark carcinomas that are less responsive to tamoxifen and more likely to progress than would be predicted by more traditional methods of ER analysis.

Application of photoshop-based image analysis to quantification of hormone receptor expression in breast cancer.

The benefit of quantifying estrogen receptor (ER) and progesterone receptor (PR) expression in breast cancer is well established. However, in routine breast cancer diagnosis, receptor expression is often quantified in arbitrary scores with high inter- and intraobserver variability. In this study we tested the validity of an image analysis system employing inexpensive, commercially available computer software on a personal computer. In a series of 28 invasive ductal breast cancers, immunohistochemical determinations of ER and PR were performed, along with biochemical analyses on fresh tumor homogenates, by the dextran-coated charcoal technique (DCC) and by enzyme immunoassay (EIA). From each immunohistochemical slide, three representative tumor fields (x20 objective) were captured and digitized with a Macintosh personal computer. Using the tools of Photoshop software, optical density plots of tumor cell nuclei were generated and, after background subtraction, were used as an index of immunostaining intensity. This immunostaining index showed a strong semilogarithmic correlation with biochemical receptor assessments of ER (DCC, r = 0.70, p < 0.001; EIA, r = 0.76, p < 0.001) and even better of PR (DCC, r = 0.86; p < 0.01; EIA, r = 0.80, p < 0.001). A strong linear correlation of ER and PR quantification was also seen between DCC and EIA techniques (ER, r = 0.62, p < 0.001; PR, r = 0.92, p < 0.001). This study demonstrates that a simple, inexpensive, commercially available software program can be accurately applied to the quantification of immunohistochemical hormone receptor studies.

Expression of p53, glutathione S-transferase-pi, and Bcl-2 proteins and benefit from adjuvant radiotherapy in breast cancer.

In clinical breast cancer research, the utility of certain biomarkers as predictors of response to surgery, chemotherapy, or hormonal therapy has been studied intensively. Much less research has been done on the relevance of biologic predictors of response to radiotherapy, which represents an effective local-regional treatment for breast cancer. The utility of biomarkers involved in DNA damage repair (p53 protein), control of programmed cell death (p53 and Bcl-2 proteins), and cellular detoxification (glutathione S-transferase-pi [GST-pi] enzyme) in predicting local breast cancer recurrence was analyzed retrospectively in two cohorts of breast cancer patients. These patients had had no detectable metastases in the axillary lymph nodes (i.e., node-negative) or in distant sites and had had similar distributions of clinicopathologic and biologic prognostic features. They had been treated by conservative surgery alone (139 case patients) or by conservative surgery followed by adjuvant radiotherapy (496 case patients) during the period from 1984 through 1990. The expression of the p53, GST-pi, and Bcl-2 proteins in the specimens of primary breast tumor obtained from these patients was determined by use of immunohistochemistry; cell proliferation activity and levels of steroid receptors were determined by use of a [3H]thymidine-labeling index assay and the dextran-coated charcoal technique, respectively. The median time of follow-up of patients was 6 years. In the analyses of patient outcomes, only local failures that presented as first events were considered. After surgery alone, the risk of local recurrence at 6 years was higher for patients with tumors exhibiting elevated levels of p53 and GST-pi protein expression than for patients with low levels (hazard ratio [HR] = 3.1, 95% confidence interval [CI] = 1.3-7.7, two-sided P = .012; HR = 2.7, 95% CI = 1.1-6.4, two-sided P = .026, respectively). Weak or no observable expression of Bcl-2 protein was only suggestive of a higher frequency of local failures. Adjustment for patient age, tumor size, cell proliferation, and estrogen receptor status did not change these findings. Conversely, in the series of patients given conservative surgery followed by radiotherapy, there was no difference in local tumor recurrence between patients with tumors expressing or not expressing each of the three markers. Our study provides indirect evidence of a benefit from radiation therapy in preventing local breast cancer relapse, particularly among node-negative patients with tumors that express elevated levels of the p53 or GST-pi proteins or that express little or no Bcl-2 protein.

Steroid Hormone Receptor Content and Lymph Node Status in Endometrial Cancer

The purpose of this study was to correlate the steroid hormone receptor status in endometrial adenocarcinoma with tumor metastasis to the pelvic and para-aortic lymph nodes, and with other known prognostic variables which influence survival. Tumor samples from 85 patients with endometrioid adenocarcinoma, or adenocarcinoma with squamous differentiation of the endometrium who underwent complete surgical staging, were assayed for cytoplasmic steroid hormone receptors using a dextran-coated charcoal technique. Steroid hormone receptor content was correlated to lymph node status, along with other prognostic variables, such as patient's age, depth of myometrial invasion, tumor grade, and pelvic cytology. By univariate analysis, the likelihood of nodal involvement was associated with younger age and poorly differentiated tumors. Multivariate analysis revealed that age, tumor grade, and myometrial involvement added significant independent prognostic information. Estrogen or progesterone receptor content did not add independent prognostic information concerning lymph node status once other factors were controlled. Knowledge of estrogen and progesterone receptor binding status in adenocarcinoma of the uterus does not replace the prognostic information imparted by careful sampling of lymph nodes.

Prognostic value of steroid hormone receptors for 5-year survival in stage II endometrial cancer.

In a prospective study, the 5-year survival of 57 patients with Stage II endometrial cancer was correlated to the estrogen (ER) and progesterone (PR) receptor concentrations using the dextran-coated charcoal (DCC) technique. The cutoff level was 3 fmol/mg of protein for ER and 6 fmol/mg of protein for PR. An histopathologic examination of the tumor tissue samples before treatment demonstrated that 16% of the cases (9 patients) were well-differentiated (Grade 1), 45% (26 patients) moderately differentiated (Grade 2), and 39% (22 patients) poorly differentiated (Grade 3). Only 3 were ER negative, whereas 19 were PR negative. The mean concentrations for ER and PR were 106 and 162 fmol/mg of protein, respectively. All patients received standard treatment, including preoperative irradiation. The total 5-year survival rate was 82% (47 of 57 patients). Deep myometrial invasion was important in Grade 3, but not in Grades 1 and 2; in these two grades, all five patients with deep infiltration survived. There was a correlation between receptor concentration and grade, with a significant difference between Grades 1 and 2 versus Grade 3. All patients who were ER negative survived, as did all patients who were PR negative in Grades 1 and 2. Of the patients with Grade 3 disease who died, four of six (67%) were PR negative. Of the patients with Grade 3 disease who survived, 10 of 16 (63%) were PR negative. Patients who survived for 5 years did not have significantly different ER and PR concentrations than those who died. The mean ER and PR concentrations in patients who survived were 99 and 159 fmol/mg of protein, respectively, compared with 108 and 178 fmol/mg of protein, respectively, for those who died.

Comparison of Tritiated Estradiol and Tamoxifen Aziridine for Measurement of Estrogen Receptors in Human Breast Cancer Cytosolss

We examined the estrogen receptor measurement in 265 human breast cancer cytosols by using a specific method based on [3H]tamoxifen aziridine labeling, sequential immunoadsorption with an antiestrogen receptor monoclonal antibody (H-222), sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and autoradiography. These new tools of molecular endocrinology revealed an impressive estrogen receptor molecular polymorphism. Given the recent finding of a similar estrogen receptor polymorphism at the messenger RNA level by several laboratories, it is tempting to speculate about its possible biological significance. To gain insight into the potential clinical relevance of this polymorphism in terms of breast cancer hormone dependence, we compared the 265 cytosols for their [3H]tamoxifen aziridine- and [3H]estradiol-binding capacities using the above-mentioned method and the conventional dextran-coated charcoal assay. We failed to identify a specific [3H]tamoxifen aziridine electrophoretic pattern with respect to the tumor estrogen receptor content as measured by the dextran-coated charcoal assay. However, an excellent correlation overall was found between the intensities of both labeling methods. Some tumors were positive for only one of these two ligands. It will be clinically important to see whether the tumors positive for [3H]tamoxifen aziridine only correspond to the small subset of tumors (10%) which respond to tamoxifen treatment despite very low estrogen receptor levels, as measured by the dextran-coated charcoal technique.

Progesterone receptor in hepatocellular carcinoma. Correlation with androgen and estrogen receptors

Progesterone receptors (PgR), estrogen receptors (ER), and androgen receptors (AR) were assayed consecutively for hepatocellular carcinoma (HCC) that was surgically removed from 19 men and three women. The methods of receptor assay were the enzyme immunoassay (EIA) for PgR and the dextran-coated charcoal (DCC) technique for ER and AR. The patients ranged in age from 32 to 77 years (average, 60.3 years). No patients had received any specific anti-cancer therapy before tissue collection. All patients but one had underlying liver disease: cirrhosis in 13 and chronic hepatitis in eight. The positive rate of each receptor was 18% for PgR, 48% for ER, and 82% for AR. The titer was highest for AR, intermediate for ER, and lowest for PgR. The titers of PgR in four PgR-positive patients ranged from only 1.1 to 3.0 fmol/mg of protein. There was no relationship between PgR, ER, and AR in terms of positivity and titer. Also, other clinical and histopathologic data did not influence the positivity or concentration of these three sex hormone receptors. It can be concluded that no or little PgR exists in the cytosol of untreated HCC.

Steroid hormone receptor levels and adjuvant tamoxifen in early breast cancer

Ten year disease-free survival (DFS) results of the Naples randomized trial of adjuvant tamoxifen (TM), 30 mg per day for 2 years versus no therapy according to receptor levels, are reported. From Feb. 1, 1978, through Dec. 31, 1983, 308 pre- and postmenopausal patients with early breast cancer entered the trial. Estrogen receptor (ER) data were available on 239 (77.6%) patients, progesterone receptor (PgR) data on 194 (63.0%), and both receptor data on 181 (58.8%). ER and PgR were assayed by dextran-coated charcoal technique in a single laboratory. The effect of adjuvant TM was significantly related to ER and PgR concentration of the primary tumor. The greatest TM benefit on DFS was evident in patients with the highest levels of receptors. The interaction between the treatment effect and receptor concentration was found whether ER and PgR were considered separately or together.

A New and Convenient Assay of Growth Hormone Binding Protein Activity in Human Serum

The recent discovery of a GH-binding protein (GH-BP) in human serum has important implications for the distribution, metabolism, and physiological activity of human GH (hGH). In this report we describe an accurate, simple, and rapid assay for measuring GH-BP in human serum, based on the dextran-coated charcoal technique for separating bound from free hGH. The present method reduces the degree of dissociation of the GH-BP complex during the separation procedure, thus increasing accuracy, allows routine use of small working volumes of sera, and can be practically used for running large scale assays. Binding of [125I]hGH to human serum GH-BP, identified and characterized by this procedure, was time, temperature, and dose dependent; the binding was highly specific for hGH and with a high affinity (Ka, 1.08 +/- 0.19 x 10(9) L/mol). Preliminary results, expressed as percent specifically bound [125I]hGH corrected for endogenous hGH, indicate that the mean GH-BP activity in 50 microL sera from normal adult men and women was 11.32 +/- 0.45%. Significantly lower values were obtained in sera of infants, 3 days after birth (1.65 +/- 0.15%), and in short-statured children with normal GH secretion (7.38 +/- 0.22%). In patients with liver cirrhosis GH-BP levels decreased to 5.93 +/- 1.14%. The availability of a simple and convenient procedure for large scale determination of GH-BP coupled with the suggestion that this protein can reflect the GH receptor provides us with an indirect means for assessing changes in GH receptor activity in various physiological and pathological conditions.

Androgen receptors in laryngeal carcinoma

Specific androgen receptors were assayed from the cytosol fraction of laryngeal carcinomas from 15 male patients. Specimens also included the non-malignant laryngeal tissues from 3 of these patients. A specific radioligand was used and involved (3H) dihydrotestosterone incubation and a dextran-coated charcoal technique. Twelve of the laryngeal carcinomas showed detectable levels of high affinity but limited capacity androgen receptors. The number of binding sites varied from zero to 23.3 fmol/mg cytosol protein, with the dissociation coefficient (Kd) ranging from 0.46 to 1.85 x 10(-9) M. Androgen receptors were also found in all three non-malignant laryngeal tissues. Binding sites for these latter androgen receptors were 4.09 to 8.16 fmol/mg cytosol protein, with a Kd of 0.81 to 0.35 x 10(-9) M. Although further studies are necessary, the presence of androgen receptors in laryngeal carcinomas suggests that antiandrogen therapy might be used in the future for treating such disease.

Circadian variations of serum sex hormone binding globulin binding capacity in normal adult men and women

Diurnal variations of serum sex hormone binding globulin (SHBG), testosterone (T) and estradiol (E2) in five normal adult men and five normal adult women were investigated. SHBG binding capacity was measured by both polyacrylamide gel electrophoresis and dextran-coated charcoal technique (DCC); T and E2 were assayed by RIA and free T and free E2 were determined by means of equilibrium dialysis. In male subjects the variations of SHBG binding capacity was associated with the changes of total T, free T and T/SHBG index, which had the highest concentrations in the morning and the lowest levels in the evening during the 24 h test period, but percentage free T remained unchanged. Serum protein concentrations did not change significantly during 24 h. No significant diurnal changes of SHBG binding capacity, total E2, free E2, percentage free E2 and percentage free T were found in female subjects in the mid-luteal phase of the menstrual cycle, although significant fluctuations of total T, free T and T/SHBG index were observed throughout the day. The results suggested that SHBG may play a buffer role in the presence of fluctuations of testosterone production during 24 h period, allowing stabilization of a bioactive fraction of the hormone both in normal adult male and female. However, the concentrations of T in normal adult women may be too low to drive any change of SHBG levels while there were no significant variations ofE2 throughout a day in the mid-luteal phase of the menstrual cycle.

Evidence of 1,25-dihydroxyvitamin D 3-receptors in human digestive mucosa and carcinoma tissue biopsies taken at different levels of the digestive tract, in 152 patients

Epidemiological and experimental data suggest that dietary calcium and 1,25-dihydroxy vitamin D 3 (1,25-(OH) 2D 3) are protective against colorectal cancers, while their activity on colon mucosa still remains unknown. Since the presence of receptors is required for steroid action, specific 1,25-(OH) 2D 3 receptors were investigated in biopsies taken at different levels of the digestive tract from the oesophagus to the rectum and in pancreas. The total study involved biopsies from 152 patients. In 82% of the cases they were paired biopsies in adenocarcinoma tissue and in adjacent normal mucosa (NM). There were 120 operated on for colorectal adenocarcinoma (HCRA). 1,25-(OH) 2D 3 receptor was assayed in tissue extract by the dextran-coated charcoal (DCC) technique and also characterised by sucrose density gradient centrifugation. Scatchard analyses showed a single class of specific high affinity-low capacity sites binding for 1,25-(OH) 2D 3 with a K d = 1.48 ± 0.8 · 10 −10 M ( n = 119). The sedimentation coefficient of the steroid receptor complex was approximately 3.2 S. The incidence of 1,25-(OH) 2D 3 receptors was significantly higher in NM (82.5%) than in HCRA (34.5%). In HCRA this incidence decreased from right colon (64.7%) to left colon (27.7%) and rectum (15%). All positive HCRA in left colon and rectum ( 16 76 ) were histologically well differentiated. The receptor content in NM and HCRA was in the same range: (median) 10–314 (58) and 13–175 (64) fmol/mg protein. These data suggest that 1,25-(OH) 2D 3 may modulate calcium transport in colon, as in the intestine. Also, loss of receptivity to 1,25-(OH) 2D 3 is observed as associated with malignant transformation of the human colorectal mucosa.

Characterization of Laboratory Working Standard for Quality Control of Immunometric and Radiometric Estrogen Receptor Assays. Clinical Evaluation on Breast Cancer Biopsies

The objective of the study was to characterize a low-cost and reliable working standard material for quality control of estrogen receptor (ER) determination with dextran-coated charcoal (DCC) and enzyme immunoassay (EIA) methods. Human fibromatous uterine lyophilized cytosol demonstrated good characteristics of stability and applicability for this purpose. Eleven laboratories participated in the intralaboratory and interlaboratory quality control study, and they achieved slightly higher coefficients of variation for ER-EIA (interlaboratory, 37.7%; intralaboratory, 22.9%) than for ER-DCC (interlaboratory, 24.2%; intralaboratory, 15.7%). There was an excellent correlation between ER results with ER-EIA and ER-DCC for 268 breast cancer biopsies. Quality assurance for ER assays using DCC techniques and immunometric methods with monoclonal antibodies (ER-EIA) can be set up with this available material of human origin to satisfy the characteristics of both techniques and the species specificity of monoclonal antibodies.

Immunohistochemistry of Estrogen Receptor Protein in Paraffin Sections: Effects of Enzymatic Pretreatment and Cobalt Chloride Intensification

Monoclonal antibodies provide important tools for the demonstration of estrogen receptors (ERs) in cases of breast cancer. This study reports an improved immunohistochemical method for the demonstration of ER in formalin-fixed paraffin-embedded tissue samples using the Abbott monoclonal antibody to ER protein. Tissue sections were pretreated briefly with trypsin, followed by DNase before the performance of the immunohistochemical reaction and cobalt chloride was used to intensify the color of the diaminobenzidine reaction product. In 20 cases, the results in paraffin sections were compared with biochemical assays with the dextran-coated charcoal technique or with immunohistochemistry performed on frozen sections. There was an excellent correlation between the results obtained with all three methods. The introduction of cobalt chloride into the chromogen solution significantly increased the sensitivity of this approach as compared with the use of diaminobenzidine alone.

Comparison of enzyme immunoassay and dextran-coated charcoal techniques for progesterone receptor determination in human breast cancer cytosols

We have compared a new enzyme immunoassay for progesterone receptors (Abbott PgR-EIA, monoclonal) with our standard radioligand binding-assay (PgR-RBA). For both assays cytosols were freshly prepared from human breast cancer specimens that had been stored frozen for up to 1 year, and predominantly tissues were used which contained critically low amounts of progesterone receptors. A highly significant correlation was observed between the PgR-EIA and PgR-RBA ( Spearman: Rs = 0.85, n = 100). Using a cut-off point of 10 fmol PgR/mg protein, 87% of the values were in accordance with each other (52% negative, and 35% positive in both assays), whereas 13% scored positive (median: 16, range 11–38 fmol PgR/mg protein) in one assay and negative (median: 7, range 0–10 fmol PgR/mg protein) in the other. Also in cytosols from human ovarian and endometrial carcinoma tissues immunoreactive PgR could be detected, and significant correlations with PgR-RBA were observed ( Rs = 0.94, n = 6) for both tissues.

Relevance of cell kinetics to hormonal response of receptor-positive advanced breast cancer.

The relationship between cell kinetics and hormonal status and the relevance of the cell kinetic variable on success of hormone-therapy in estrogen receptor positive (ER+) breast tumors were analyzed in patients with advanced disease. Cell kinetics were evaluated as in vitro 3H-thymidine labeling index (LI), and estrogen receptor (ER) and progesterone receptor (PgR) with the dextran-coated charcoal technique. The analyses performed on primary tumor or soft tissue metastases from 52 patients showed a general association between the presence of hormone receptors and low proliferative activity, or the absence of receptors and high proliferative activity (ER and L.I.: p greater than 0.05; PgR and L.I.: p = 0.05). However, hormonal status and cell kinetic status were unrelated in about 40% of the cases. Clinical response to additive hormonotherapy was analyzed in relation to pretreatment LI in 29 patients with ER+ tumors. Time to reach maximum response was significantly longer in slow than in fast proliferating tumors, but complete remission was reached in 88% of slow proliferating tumors compared to only 46% of fast proliferating tumors. These preliminary results show that ER+ fast proliferating tumors largely escape hormonal control, and if confirmed on larger series, could identify cell kinetics as an important tool to select patients who will benefit from hormonal treatment.

Cytoplasmic steroid receptors in cancer of the larynx.

There are many data that suggest that the larynx is a target organ for steroid hormones, especially androgens. Since laryngeal cancers occur much more often in males, it is conceivable that androgens might be the growth-stimulating factor for such tumors. In order to test this concept further, neoplastic tissues from 68 patients (64 males, 4 females) were obtained during planned total laryngectomies and were tested for the presence of androgen (AR) and estrogen (ER) receptors. In 18 cases progesterone receptors (PgR) were also quantified. The dextran-coated charcoal technique was used to determine hormone receptors and the number of the binding sites and the equilibrium constant of dissociation were calculated according to Scatchard. 3H-R1881 was used as the ligand for the measurement of AR, 3H-estradiol for ER, and 3H-R5020 for PgR. AR was present in 11 tumor specimens (1 female, 10 males) and the concentration varied from 2.8 to 17.1 fmol/mg proteins. ER were found in 6 tumors (1 female, 5 males) in concentrations of 2.9-11.2 fmol/mg proteins. Three tumor specimens contained both AR and ER. All tumors analyzed lacked PgR. These results suggest that at least some laryngeal carcinomas might be hormone dependent, indicating that certain patients could benefit from antiandrogen therapy.

Correlation between ERICA and DCC Assay in Hormone Receptor Assessment of Human Breast Cancer

To study the basic relationships between estrogen receptor immunocytochemical assay (ERICA) and dextran-coated charcoal (DCC) techniques, 116 women affected by breast cancer were admitted to the study from June 1985. In 56% of cases the tumor sample came from patients with operable disease and in 44% from patients with advanced inoperable disease. We found an overall agreement of 80% between the two methods characterized by a high sensitivity (95%) and a low specificity of ERICA versus DCC. ER/ICA negative results were found in 20 out of 78 ER/DCC positive cases; on the contrary, discordant results were found in only 3 out of 38 cases described by DCC as ER negative. 29 patients with advanced disease and ER/DCC positive tumors were considered for clinical analysis and reliability of each hormone receptor assay. A clinical response was reached in 69% of ER/DCC positive patients and on 64% of ER/ICA positive ones (16/25). However, it must be noted that 4/29 cases described as negative by ERICA were clinically responsive to hormone therapy.