estradiol-6-(o-carboxymethyl)oxime and Breast-Neoplasms

According to the character of structure and activity of the test reagents I-IV, two new reagents V and VI for detecting estrogen receptor of human mammary cancer cells were synthesized. This simplifies the route of synthesis and increases activity. Key intermediates VIII and IX were confirmed by IR, MS, UV and elemental analysis. The quantification of the final products V and VI were determined by UV. The result of preliminary clinico-pathological test shows compound V to be effective on estrogen receptor.

Topics: Breast Neoplasms; Estradiol; Estrogens, Conjugated (USP); Ethinyl Estradiol; Humans; Reagent Kits, Diagnostic; Receptors, Estrogen

The performance of a fluorescent cytochemical oestrogen receptor (ER) assay has been compared with an established dextran-coated charcoal ER assay in 85 cases of primary breast cancer. Both assays were performed on tissue samples from the same tumour, by independent observers in different laboratories. A good qualitative correlation between the two methods was found, with agreement in 92 per cent of cases (P less than 0.0005). Criticisms of the fluorescein-labelled oestrogen methods are discussed. It is concluded that such methods should be validated in each laboratory by comparison with a conventional, quantitative ER assay before being used for clinical decisions.

Topics: Adsorption; Adult; Aged; Breast Neoplasms; Charcoal; Estradiol; Female; Fluorescein-5-isothiocyanate; Fluoresceins; Histocytochemistry; Humans; Methods; Microscopy, Fluorescence; Middle Aged; Receptors, Estrogen; Serum Albumin, Bovine; Thiocyanates

The present study defines criteria for determining the presence of estrogen-receptors in human breast carcinomas demonstrated by a histochemical assay using 17 beta-estradiol-carboxy-methyl-oxim-bovine serum albumen-FITC. The criteria were: 1) the percentage of cells showing fluorescence; 2) the intensity of the fluorescence observed, and 3) the percentage of epithelial structures in tissue specimens. Using these predefined criteria in 132 human breast carcinomas as 91.6% agreement was found between the results of the histochemical assay and those of the biochemical Charcoal method. The main causes of disagreement (7 of the 11 cases) were sampling errors between the tissue specimens used for the histochemical and biochemical assay, and an insufficient percentage of epithelial structures (less than 15%) to allow biochemical identification of estrogen receptor activity. In the hands of pathologists with experience of the field of histochemistry this histochemical assay may be the method of choice for the assessment of estrogen receptors.

Topics: Adult; Aged; Animals; Breast Neoplasms; Cattle; Cell Transformation, Neoplastic; Epithelium; Estradiol; Fluorescein-5-isothiocyanate; Fluoresceins; Histocytochemistry; Humans; Middle Aged; Receptors, Estrogen; Serum Albumin, Bovine; Thiocyanates

Three histochemical techniques for estrogen binding localization in tissue sections were compared to sucrose density gradient analyses for estrogen receptor in 186 breast carcinomas. Apparent localization to epithelial elements was noted using 6-BSA-Fluor-CMO-17 beta-estradiol, 17-BSA-Fluor-TSC-estrogen, and polyestradiol phosphate/anti-estradiol antibody methods. The correlation between the histochemical methods and standard sucrose density gradient techniques was poor for the polyestradiol antibody method and the 6-BSA-fluor-CMO-17 beta-estradiol techniques. While an improved correlation was observed with the 17-BSA-Fluor-TSC-estrogen compound, the compound's binding was not effectively blocked by preincubation with diethylstilbestrol. This failure to show convincing saturability of binding, combined with problems of extraction of soluble receptor proteins in the aqueous incubation media and washed from cells rendered permeable by cryostat sectioning, indicates that several areas will require clarification before histochemical techniques can begin to be considered as a method for estrogen ""receptor" analyses in the clinical evaluation of breast neoplasms.

Topics: Binding, Competitive; Breast Neoplasms; Centrifugation, Density Gradient; Estradiol; Estrogens, Conjugated (USP); Female; Histocytochemistry; Humans; Oximes; Receptors, Estrogen; Structure-Activity Relationship