Journal Article Immunohistochemical Progesterone Receptor Assay: Measurement by Image Analysis Get access Nafissa El-Badawy, M.D., Nafissa El-Badawy, M.D. Department of Pathology and Laboratory Medicine, Emory University School of Medicine, Atlanta, Georgia. Address reprint requests to Dr. Cohen: Anatomic Pathology, Emory University, 1364 Clifton Road, NE, Atlanta, Georgia 30322. Search for other works by this author on: Oxford Academic Google Scholar Cynthia Cohen, M.D., Cynthia Cohen, M.D. Department of Pathology and Laboratory Medicine, Emory University School of Medicine, Atlanta, Georgia. Search for other works by this author on: Oxford Academic Google Scholar Patricia B. Derose, MT(ASCP)SH, Patricia B. Derose, MT(ASCP)SH Department of Pathology and Laboratory Medicine, Emory University School of Medicine, Atlanta, Georgia. Search for other works by this author on: Oxford Academic Google Scholar Irene J. Check, PH.D., Irene J. Check, PH.D. Department of Pathology and Laboratory Medicine, Emory University School of Medicine, Atlanta, Georgia. Search for other works by this author on: Oxford Academic Google Scholar Demetrios Sgoutas, PH.D. Demetrios Sgoutas, PH.D. Department of Pathology and Laboratory Medicine, Emory University School of Medicine, Atlanta, Georgia. Search for other works by this author on: Oxford Academic Google Scholar American Journal of Clinical Pathology, Volume 96, Issue 6, 1 December 1991, Pages 704–710, https://doi.org/10.1093/ajcp/96.6.704 Published: 01 December 1991 Article history Received: 15 December 1989 Accepted: 15 January 1991 Published: 01 December 1991
Image cytometric quantitation of nuclear DNA of paragangliomas may provide prognostic information that cannot be obtained from histopathologic study. Flow cytometry has demonstrated DNA aneuploid tumors to have a higher risk of progression than diploid neoplasms.DNA ploidy of 56 paragangliomas was assessed by image cytometry of 5-micron, Feulgen-stained, formalin-fixed, paraffin-embedded tissue sections.Thirty-three (59%) paragangliomas were diploid and 23 (41%) aneuploid. Of the 30 adrenal pheochromocytomas, 15 (50%) were diploid. Thirteen (93%) of the 14 carotid body tumors were diploid. Five of seven (71%) glomus jugulare tumors and two of five (40%) extraadrenal paragangliomas were aneuploid. During a mean follow-up of 57 months (range, 1 month to 36 years) of 44 patients with 47 paragangliomas, 33 (75%) were alive and without disease; 7 (16%), including 1 glomus jugulare, 2 carotid body and 4 pheochromocytoma patients, developed recurrences/metastases. By multivariate analysis, image cytometric DNA ploidy was predictive of disease-free survival for adrenal pheochromocytomas. No significant differences in overall survival, disease-free survival or recurrence/metastasis rate were noted between other diploid and aneuploid tumors.Aneuploidy suggests a risk of early recurrence for adrenal pheochromocytomas.
To define the histogenesis of the Paget cell and possibly identify differences in cells from the two sites, six vulvar and 23 mammary specimens from Paget's disease lesions were studied for immunocytochemical antigens. All vulvar and 21 (91%) mammary lesions were strongly reactive for glandular cytokeratin. All lesions showed immunopositive Paget cells with epithelial membrane antigen (EMA). An apocrine antigen, gross cystic disease fluid protein (GCDFP-15), decorated 66.5% and 56.5% of extramammary and mammary lesions, respectively. All vulvar Paget cells stained for carcinoembryonic antigen (CEA), a frequency significantly greater than the 35% in mammary lesions (p = 0.02). However, CEA is expressed by both eccrine and apocrine sweat glands and their tumors. Vulvar Paget cells were negative for lysozyme, casein, lactalbumin, and S100 protein, compared with 9%, 4%, 4%, and 26% in nipple lesions. S100 protein expression is similar to that in mammary ductal carcinoma (32%). The glandular origin of both extramammary and mammary Paget cells is indicated by the presence of glandular cytokeratin, EMA, and CEA. Approximately 60% of all cases in both sites showed evidence of apocrine derivation (GCDFP-15 positivity). Variable antigen expression suggests possible malignant transformation of pluripotent germinative cells able to differentiate in an apocrine or an eccrine direction, or in both.
Ductal carcinoma in situ of the breast (DCIS) was studied for frequency of biologic markers. Sections were immunostained for the presence of estrogen receptor (ER), progesterone receptor (PR), c-erbB-2, epidermal growth factor receptor (EGFR), cathepsin D, and p53 with the TechMate automated immunostainer, using an avidin-biotin technique and microwave antigen retrieval. Feulgen-stained and immunostained sections were assayed for DNA ploidy and for proliferative activity (MIB-1) and angiogenesis (Factor VIII-related antigen [FVIII), respectively, using the CAS 200 image cytometer. Of the 74 DCIS, 22% were comedocarcinomas, 10% high grade, and 18% nuclear poorly differentiated. ER was present in 72%, PR in 86%, c-erbB-2 in 34%, EGFR in 0%, cathepsin D in 43%, and p53 in 57%. Sixty-five percent were aneuploid and 65% had proliferative activity (mean, <9% MIB-1); 58% had angiogenesis (mean, <6.2% FVIII). There was no significant difference in frequencies of any of these parameters according to classification except for aneuploidy and nuclear differentiation (p = 0.04). Eighty-seven percent were treated by modified radical mastectomy and 13% by local resection only. In a 6.2-year mean follow-up after mastectomy (range, 1 month to 9 years), there were no recurrences or deaths due to disease. Longer follow-up after mastectomy and follow-up of patients after lumpectomy with and without radiation is warranted.
Estrogen receptor (ER) positivity demonstrated in malignant melanomas by histochemical and biochemical assays suggested the possibility of hormonal management and improved prognosis as for breast carcinoma patients. We studied the ER status of 5 primary and 28 metastatic malignant melanomas with a commercial immunohistochemical kit (ER-ICA monoclonal), that utilizes monoclonal anti-ER and a peroxidase-antiperoxidase technique, and by a histochemical method using fluorescein-conjugated estradiol (Fluoro-Cep Estrogen assay), on frozen sections. In addition, we conducted a biochemical assay [dextran-coated charcoal cytosolic assay (DCC)] in 16 cases. All 33 cases were ER negative by ER-ICA and Fluoro-Cep: 11 biochemical assays were negative (less than 3 fmol ER/mg protein), four were in the borderline range (3 to 10 fmol ER/mg protein), and one was positive (greater than 10 fmol ER/mg protein) at 11 fmol. The melanomas in 97% of the cases we studied were ER negative by two or three different assays. Low-level estrogen binding of MM tissues may be the result of interactions other than with Type I true ER. The low frequency of ER positivity of malignant melanomas appears to preclude the clinical use of ER status as an indicator for response to hormonal manipulation in patients with malignant melanoma.
