Detection and dynamic localisation of estradiol-receptor complexes in intact target cells by immunofluorescence technique

Nenci, Italo; Beccati, Maria Donatella; Piffanelli, A; Lanza, Giovanni

doi:10.1016/0022-4731(76)90194-1

Cited by 89 publications

(23 citation statements)

References 20 publications

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“…In our cases we did not find nuclear posi tivity [29,30] for estradiol as reported in target tissues [29,30], therefore the cytoplas matic positivity could be an expression of endogenous estradiol produced ex novo by the neoplastic cells [31] or diffused into the cytoplasm across the cytoplasmatic mem brane as we reported in cutaneous melano mas [28]. Alternatively the cytoplasmatic positivity could be an estradiol-receptor com plex that cannot be activated or translocated [29,30].…”

Section: Discussionmentioning

confidence: 59%

Steroids and Neuroendocrine Hormones Detected by the Immunoperoxidase Technique from Malignant Melanomas and Nevi of the Choroid and Conjunctiva

Tondo¹,

Balestrazzi

Santeusanio³

et al. 1986

Ophthalmologica

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By the peroxidase-antiperoxidase technique, the authors studied 7 malignant choroidal melanomas, 7 conjunctival nevi and 1 malignant conjunctival melanoma with the aim to detect the presence of vasoactive intestinal polypeptide (VIP), adrenocorticotropic hormone (ACTH), gastrin, estradiol and testosterone. Positive staining reaction for VIP, estradiol and testosterone was observed in both malignant melanomas of the choroid and conjunctival nevi. The case of conjunctival melanoma was positive for VIP and ACTH but not for estradiol and gastrin.

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Section: Discussionmentioning

confidence: 59%

Steroids and Neuroendocrine Hormones Detected by the Immunoperoxidase Technique from Malignant Melanomas and Nevi of the Choroid and Conjunctiva

Tondo¹,

Balestrazzi

Santeusanio³

et al. 1986

Ophthalmologica

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“…Numerous studies have documented the progressive loss of activity of labile molecules after the surgical interruption of blood flow. [16][17][18][19][20] The standardization of cold ischemic time is important and is a modifiable factor in proper assessment of biomarkers. The pathologist should effectively communicate this priority to all members of the breast care management team so processes are put in place to make sure the time of tissue collection, and the fixation start time is routinely recorded.…”

Section: Discussionmentioning

confidence: 99%

“…15 The progressive loss of activity of labile macromolecules after the surgical interruption of blood flow leading to tissue ischemia, acidosis, and enzymatic degradation has been documented. [16][17][18][19][20] The warm and cold ischemic times are accepted as important variables in the analysis of these macromolecules from clinical tissue samples. While it is not possible to control the warm ischemic time, cold ischemic time can be controlled with proper specimen handling.…”

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confidence: 99%

The effect of cold ischemic time on the immunohistochemical evaluation of estrogen receptor, progesterone receptor, and HER2 expression in invasive breast carcinoma

2012

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The American Society of Clinical Oncology/College of American Pathologist (ASCO/CAP) guidelines on breast cancer currently recommend a cold ischemic time of o1 h, although data are limited. Breast resection specimens were subjected to variable cold ischemic time periods (0.5, 1, 2, 3, 4, 24, and 48 h) within the refrigerator and at room temperature. The study included 25 tumors, all of which had refrigerated samples. Nonrefrigerated samples (samples at room temperature) were present on 23 cases. Hormone receptors were semiquantitatively scored using the H-score method. Human epidermal growth factor receptor 2 gene ERBB2 (HER2) was scored using the ASCO/CAP guidelines. The results were compared with the core biopsy scores, which have negligible cold ischemic time period before fixation. Mild reduction in staining for hormone receptors was judged present if the H-score on the resection specimen was between one-half and three-fourth of the H-score at core biopsy. Significant reduction was judged present when the H-score on resection was less than one-half of the core biopsy H-score. Mild reduction in HER2 staining was judged present if there was one-step reduction; and significant reduction was judged present if there was two-step reduction in staining. A true reduction was judged present only when the reduction was consistently present for the increasing time interval. A focal reduction for a particular time sample was attributed to the heterogeneity of the tumor sample. Non-refrigerated samples are affected more by prolonged cold ischemic time than refrigerated samples. Cold ischemic time period of as short as one-half hour may occasionally impact the immunohistochemical (IHC) staining for progesterone receptor. Significant reduction in IHC staining for hormone receptors, and HER2, however, generally does not result until 4 h for refrigerated samples and 2 h for non-refrigerated samples. The ASCO/CAP guideline of cold ischemic time period of o1 h is a prudent guideline to follow. Modern Pathology (2012Pathology ( ) 25, 1098Pathology ( -1105 doi:10.1038/modpathol.2012 published online 30 March 2012 Keywords: breast carcinoma; cold ischemic time; ER/PR/HER2 All invasive primary breast cancers are analyzed for at least estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor 2 gene ERBB2 (HER2), as these markers have been shown to be the most important prognostic and predictive factors in breast cancer. The patients with ER þ primary invasive breast cancer derive considerable benefit from endocrine therapy, while ER-negative tumors do not. Similarly to ER expression, PR expression by immunohistochemistry also provides prognostic and predictive information. 1,2 Accurate determination of biomarkers is essential for the proper management of breast cancer patients. [3][4][5][6][7][8] The HER2 is overexpressed in B15-20% of breast carcinomas. 9-11 HER2 positivity is associated with higher rate of recurrence and mortality in newly diagnosed breast cancer patients, and agents that t...

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“…In order to circumvent these disadvantages of the current ER assay, many immunocytochemical methods using antiestradiol antibody (7)(8)(9)(10)(11) and cytochemical methods using fluorescein-or peroxidase-labeled estradiol (12)(13)(14)(15) have been proposed. However, sucrose gradient analysis revealed that the antibodies against estradiol do not necessarily detect the estradiol-ER complex (16).…”

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confidence: 99%

Immunocytochemical staining of estrogen receptor in paraffin sections of human breast cancer by use of monoclonal antibody: comparison with that in frozen sections.

Shimada¹,

Kimura²,

Abe³

et al. 1985

Proc. Natl. Acad. Sci. U.S.A.

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Estrogen receptor (ER) in human breast cancer tissues was demonstrated in paraffin sections as well as in frozen sections by immunoperoxidase methods using monoclonal antibody (H222) against ER. The avidin-biotin-peroxidase complex method was used for the paraffin sections fixed in cold buffered formalin, and the peroxidase-antiperoxidase method was used for the fixed frozen sections. The results were compared with the ER content in the respective tumor tissue determined by dextran-coated charcoal assay. The specific staining for ER was located exclusively in the nuclei of cancer cells in both paraffin and frozen sections. Differences in the intensity and distribution of nuclear staining within a section were often observed, suggesting heterogeneity of the ER content of individual breast cancer cells. In 24 breast cancer tissues studied simultaneously by both paraffin and frozen section methods, 21 (88%) showed similar evaluation of the presence of ER. (6).In order to circumvent these disadvantages of the current ER assay, many immunocytochemical methods using antiestradiol antibody (7-11) and cytochemical methods using fluorescein-or peroxidase-labeled estradiol (12-15) have been proposed. However, sucrose gradient analysis revealed that the antibodies against estradiol do not necessarily detect the estradiol-ER complex (16). In addition, affinity of the estradiol-conjugate for ER was shown to be extremely low when compared with that of free estradiol (17)(18)(19). Although these published methods were aimed at visualizing the specific binding of estrogen to ER, the methods of tissue preparation used in these studies could not prevent the loss of ER through diffusion or the reduced ability of estrogen to bind to its receptor (20). Therefore, it is reasonable to conclude that the stains obtained by these immunocytochemical and cytochemical methods are not specific for ER (16)(17)(18)(19)(20).Recently, monoclonal antibodies to human ER were developed by Greene et al. (21) and Miller et al. (22). By use of these monoclonal antibodies against ER, King and Greene first demonstrated ER in frozen tissue sections prepared from human breast cancer and other sources (23). These monoclonal antibodies are highly specific for ER and could serve as a more reliable probe for detecting ER in tissues. However, frozen sections have limitations in their use for detailed examination of tumor morphology and for retrospective studies.In this study, methods were developed to demonstrate ER in cold formalin-fixed paraffin sections as well as in frozen sections of human breast cancer tissues by use ofone of these monoclonal antibodies. The utility of immunocytochemical staining of ER was evaluated by comparing the results with the results of determining ER content by the dextran-coated charcoal (DCC) assay. MATERIALS AND METHODSBreast Cancer Tissues. One hundred and thirteen breast cancer tumors (104 primary and 9 metastatic tumor tissues) were obtained at surgery at the National Cancer Center Hospital and Keio University Hosp...

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Detection and dynamic localisation of estradiol-receptor complexes in intact target cells by immunofluorescence technique

Cited by 89 publications

References 20 publications

Steroids and Neuroendocrine Hormones Detected by the Immunoperoxidase Technique from Malignant Melanomas and Nevi of the Choroid and Conjunctiva

Steroids and Neuroendocrine Hormones Detected by the Immunoperoxidase Technique from Malignant Melanomas and Nevi of the Choroid and Conjunctiva

The effect of cold ischemic time on the immunohistochemical evaluation of estrogen receptor, progesterone receptor, and HER2 expression in invasive breast carcinoma

Immunocytochemical staining of estrogen receptor in paraffin sections of human breast cancer by use of monoclonal antibody: comparison with that in frozen sections.

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