Criteria for the Establishment of a Double-Labeling Assay for Simultaneous Determination of Estrogen and Progesterone Receptors

Grill, H. J.; Manz, B.; Bělovský, O.; Pollow, K.

doi:10.1159/000225785

Cited by 24 publications

(8 citation statements)

References 9 publications

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“…As a first Step, we adapted double isotope methods for Simultaneous receptor determination in cytosols (19 -22) to a cytosolic assay with isoelectric focussing for receptor quantitation. The results were comparable to those described in the literature (19)(20)(21)(22). In a second Step, this method was used in the analysis of the receptor content in supernatants from frozen sections.…”

Section: Discussionsupporting

confidence: 90%

“…To further minimize the amount of tissue needed, and to determine both receptor species quantitatively and simultaneously from the same set of frozen sections, we describe an assay in which sections are incubated in the presence of an iodinated gestagenic and tritiated oestrogenic ligand and vice versa, according to assays described for cytosols (19)(20)(21)(22).…”

Section: Introductionmentioning

confidence: 99%

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Simultaneous Quantification of Oestrogen and Progesterone Receptors by a Ligand Binding Assay in Frozen Sections

Vollmer¹,

Helmchen²,

Knüppen³

1989

Clinical Chemistry and Laboratory Medicine

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Summary:We describe two modifications of a double-isotope assay for measuring the concentrations of oestrogen receptors and progesterone receptors in tumour cytosols and extracts of frozen tumour sections and endometrial sections. The concentrations of these receptors are derived from single-point/isoelectric focussing assays after incubation of cytosols or section supernatants either with ([ H]pregn-4-ene-3,20-dione). The concentrations of the oestrogen and progesterone receptors found in cytosols (r > 0.93) and extracts from sections (r > 0.8) by dual label assays are highly correlated with those found by single label assays. The method described represents an approach to the determination of oestrogen and progesterone receptors biochemically in an amount of tissue which is comparable to that needed for immunocytochemical procedures.

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Section: Discussionsupporting

confidence: 90%

Section: Introductionmentioning

confidence: 99%

Simultaneous Quantification of Oestrogen and Progesterone Receptors by a Ligand Binding Assay in Frozen Sections

Vollmer¹,

Helmchen²,

Knüppen³

1989

Clinical Chemistry and Laboratory Medicine

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“…An ER-positive breast cancer was used as positive control. ER positivity for this breast cancer sample was determined biochemically as described (17). Corresponding species-and isotype-specific IgGs were used as negative controls.…”

Section: Methodsmentioning

confidence: 99%

Estrogen Signaling Is Active in Cartilaginous Tumors: Implications for Antiestrogen Therapy as Treatment Option of Metastasized or Irresectable Chondrosarcoma

Cleton‐Jansen

Beerendonk²,

Baelde³

et al. 2005

Clinical Cancer Research

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Purpose: Chondrosarcoma is a malignant cartilaginous matrix^producing tumor that can be lethal in 10% to 50% of the patients. Surgery is the only effective treatment known as these tumors are notorious refractory to all types of conventional chemotherapy or radiotherapy. To identify a target for therapy, we want to determine whether estrogen signaling is active in chondrosarcoma because estrogen is important in the regulation of longitudinal growth that is initiated by chondrocyte proliferation and differentiation in the epiphyseal growth plate of long bones. Experimental Design: We studied protein expression of the estrogen receptor in 35 cartilaginous tumors as well as mRNA levels for the estrogen receptor and for aromatase, an enzyme for estrogen synthesis and another potential therapeutic target. Furthermore, the activity of aromatase was determined in vitro by the tritiated water release assay. Dose-response experiments with chondrosarcoma cultured cells were done with estrogen, androstenedione, and exemestane. Results: All chondrosarcomas tested showed mRNA and nuclear protein expression of the estrogen receptor. Also, aromatase mRNA was detected. The aromatase activity assay showed a functional aromatase enzyme in primary chondrosarcoma cultures and in a cell line. Growth of chondrosarcoma cell cultures can be stimulated by adding estrogen or androstenedione, which can be inhibited by exemestane. Conclusions: These results show, on the RNA, protein, and cell biological levels, that the ligand and the receptor are active in estrogen-mediated signal transduction. This observation implicates potential use of targeted drugs that interfere with estrogen signaling, such as those applied for treating breast cancer.

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“…Estrogen and progesterone receptor analysis of cytosols was performed with a dual label assay using 160 ␣-125 I-iodo-3,17-estradiol ( 125 I-E2) and 3 H-R5020 as ligands, according to the method of Grill et al (1984). Scatchard analysis was used to analyze the estrogen and progesterone binding data.…”

Section: Measurement Of Hormone Receptorsmentioning

confidence: 99%

Differential characteristics of two new tumorigenic cell lines of human breast carcinoma origin

Möbus¹,

Moll²,

Gerharz³

et al. 1998

Int. J. Cancer

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Criteria for the Establishment of a Double-Labeling Assay for Simultaneous Determination of Estrogen and Progesterone Receptors

Cited by 24 publications

References 9 publications

Simultaneous Quantification of Oestrogen and Progesterone Receptors by a Ligand Binding Assay in Frozen Sections

Simultaneous Quantification of Oestrogen and Progesterone Receptors by a Ligand Binding Assay in Frozen Sections

Estrogen Signaling Is Active in Cartilaginous Tumors: Implications for Antiestrogen Therapy as Treatment Option of Metastasized or Irresectable Chondrosarcoma

Differential characteristics of two new tumorigenic cell lines of human breast carcinoma origin

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