In situ hybridization of chromosome 6 on fine-needle aspirates from breast carcinomas: Comparison of numerical abnormalities and ER/PgR status and staining pattern

The estrogen receptor (ER) gene is located on chromosome 6. The aim of our study was to investigate whether numerical chromosomal aberrations were reflected in estrogen/progesterone receptor (PgR) status and staining pattern. Fine-needle aspirates from 51 breast carcinomas were investigated immunocytochemically for ER/PgR and by in situ hybridization technique using digoxigenin-labeled α-satellite probe for chromosome 6. Cases with ≥70% two-signal nuclei were regarded as disome; the remaining tumors showed aneusomy with a variable number of signals. Aneusomy was found in 32 tumors (63%), whereas 19 (37%) had a normal number of chromosome 6. Chromosomal gain occurred in all aneusome cases except one. ER- and/or PgR-positive tumors had an equal distribution of disomy and aneusmy. Variable ER staining pattern or ER and/or PgR negativity was associated with numerical aberrations in chromosome 6 in 76% of the tumors. Cancers with uniform ER staining pattern all had normal chromosome number. Diagn. Cytopathol. 16:420–424, 1997. © 1997 Wiley-Liss, Inc.     

Steroid hormone receptors in breast cancer

The significance of hormone receptor (estrogen(ER)- and progesterone receptor (PgR) assay was described with special reference to the treatment of advanced and early breast cancer. Fifty to sixty percent of ER-positive breast cancer responds to endocrine treatment, while only about 10% of ER-negative cancer do. Advanced breast cancer patients with ER-positive tumors survive longer than those with ER-negative tumors, mainly because of better response to therapy. The clinical benefit of assaying the hormone receptors in primary breast cancer was discussed, particularly concerning with the relapse-free and overall survivals of the patients after primary operation. The findings suggest the possibility of selecting operable breast cancer patients for the most appropriate adjuvant using ER and PgR.     

Hormone receptor status and HER2/neu overexpression determined by automated immunostainer on routinely fixed cytologic specimens from breast carcinoma: correlation with histologic sections determinations and diagnostic pitfalls

Although fine-needle aspiration cytology is a routine procedure for the diagnosis of breast carcinoma, cytologic specimens have rarely been used for evaluation of hormone receptor status and HER2/neu overexpression. In order to compare the biological markers on cytology and on histology, routinely fixed smears of 110 primary breast carcinomas were immunostained for estrogen receptor (ER), progesterone receptor (PgR), and HER2/neu by automated immunostainer and the results were compared with the corresponding histologic sections. ER was expressed in 76 of 110 (69%) cases and PgR was expressed in 51 of 110 (46%). Overexpression of HER2/neu was observed in 30 of 110 (27%) cases. Concordance between cytology and histology was 98% for ER, 95% for PgR, and 100% for HER2/neu. There was no false positive result on smears. Diagnostic pitfalls in determination of hormone receptor status on smears included intratumoral heterogeneity and presence of mucin.     

Evaluation of androgen receptor and GATA binding protein 3 as immunohistochemical markers in the diagnosis of metastatic breast carcinoma to the lung

Differentiating metastatic breast carcinoma in the lungs from primary lung tumors and mesotheliomas is important for determining prognosis and treatment. We evaluated novel breast specific markers, androgen receptor (AR) and GATA binding protein 3 (GATA3) immunohistostaining, for this differential, and compare to other traditional markers. The specimens comprised 33 metastatic breast carcinomas to the lung, 566 primary lung tumors (170 adenocarcinomas, 157 squamous cell carcinomas, 31 pleomorphic carcinomas, 115 large cell neuroendocrine carcinomas, 43 small cell carcinomas, and 49 typical carcinoids) and 42 malignant mesotheliomas. They were analyzed by immunohistochemistry using antibodies to AR, GATA3, estrogen receptor (ER), progesterone receptor (PgR), mammaglobin, gross cystic disease fluid protein‐15 (GCDFP‐15). Of the metastatic breast carcinomas, immunohistostaining of AR, GATA3, ER, PgR, mammaglobin, GCDFP‐15 were positive in 27 cases (81.8%), 24 cases (72.7%), 26 cases (78.8%), 13 cases (39.4%), 12 cases (36.4%), 9 cases (27.3%), respectively. Of primary lung tumors and mesotheliomas, staining of AR, GATA3, ER, PgR, mammaglobin, GCDFP‐15 were positive in 18 cases (3%), 3 cases (0.5%), 4 cases (0.7%), 2 cases (0.3%), 0 case (0%), 2 cases (0.3%), respectively. Immunohistochemistry of AR and GATA3 are reliable for differentiating metastatic breast carcinoma from primary lung tumors and mesotheliomas.     

DNA of mouse mammary tumor virus‐like virus is present in human tumors influenced by hormones

Mouse mammary tumor virus (MMTV) is a hormonally regulated, oncogenic virus of mice. MMTV‐like virus DNA has previously been detected in human breast cancers, liver disease, and liver cancers. It is hypothesized that local hormonal effects might be of primary importance in determining MMTV‐like virus detection in human tumors. MMTV‐like virus envelope (env) DNA was determined using nested PCR in 89 ovarian, 147 prostate, 50 endometrial, 141 skin, and 51 lung cancers. Viral‐positive sequences were compared with published MMTV‐like viral sequences from human breast cancer, liver cancer and MMTV. Immunohistochemistry for estrogen receptor (ER‐α) and progesterone receptor (PgR) was performed on a subset of tumors. MMTV‐like virus env DNA was detected in ovarian cancers (14/89; 16%), prostate cancers (53/147; 36%), endometrial cancers (5/50; 10%), skin cancers (13/141; 9%) but not in lung cancers (0/51). Phylogenetic analysis of the viral‐positive sequences showed no clustering of the isolates according to tissue type. A significant association was observed between the presence of hormone receptors and detection of MMTV‐like virus in the human cancers screened (P = 0.01). A significant association between MMTV‐like virus and PgR was noted in skin cancers (P = 0.003). Therefore, unlike the mouse model, the detection of MMTV‐like env sequences in human cancers in addition to breast indicates that MMTV‐like viral expression is not breast cancer‐specific and may relate to hormone‐dependent viral expression. J. Med. Virol. 82:1044–1050, 2010. © 2010 Wiley‐Liss, Inc.     

Immunocytochemical assay of progesterone receptors in paraffin-embedded specimens of endometrial carcinoma and hyperplasia: a preliminary evaluation

An immunohistochemical method for the detection of progesterone receptors (PgR) using the monoclonal anti-PgR antibody KD 68 was utilized to study paraffin-embedded tissue sections from women with endometrial carcinoma and hyperplasia. Stromal as well as myometrial nuclear PgR were nearly always apparent. In carcinoma, 11/24 (46%) of cases showed epithelial positivity, whereas in hyperplasia 8/9 (89%) were PgR-positive (P less than 0.05). Initial biochemical PgR assays by the dextran-coated charcoal method were compared with results of PgR-immunocytochemical assays (ICA) in the paraffin-embedded tissue and were in concordance in 92%. In the one discordant specimen, PgR-ICA-negative tumor cells were seen infiltrating PgR-ICA-positive myometrium, and the biochemical assay was thus felt to be falsely positive. Twelve additional cases of endometrial carcinoma were also studied for estrogen receptor (ER) by immunocytochemistry. Two were positive for both ER and PgR, while five were negative for both receptors. The immunocytochemical methods described allow for analysis of routinely fixed, paraffin-embedded specimens, thus permitting analysis of very small specimens and archival material.     

Immunocytochemical analysis of receptors for estrogen and progesterone in fine-needle aspirates from human breast carcinomas

Monoclonal antibodies were used for a preoperative analysis of estrogen receptors (ERs) and progesterone receptors (PgRs) in fine-needle aspirates from 44 primary human breast carcinomas. The semiquantitative receptor values obtained in cytologic specimens correlated well with those from enzyme immunoassay analysis on surgically removed tumor tissue (r = 0.74 for PgR; r = 0.75 for ER). Cytologic smears showed a heterogenous tumor cell distribution of ER and PgR in 29 and 23 cases, respectively. The results suggest that measurement of the ER and PgR in cytologic smears is an accurate and reliable technique that can be performed on a minimum amount of tissue.     

Estrogen and progesterone receptor contents in ThinPrep-processed fine-needle aspirates of breast.

This study was undertaken to assess the potential value of ThinPrep-processed (Cytyc, Boxborough, MA) smears from malignant breast fine-needle aspirates (FNAs) for the determination of estrogen receptor (ER) and progesterone receptor (PR) status. The ER and PR content of 142 malignant FNAs were compared with the results of the surgically excised tumors in which the assay was done by enzyme immunoassay in 97 cases or by immunohistochemistry in 45 cases. Monoclonal antibodies directed against ER-1D5 (Dako, Carpinteria, CA) and PR-1A6 (Dako) were used with the antigen retrieval technique. By using enzyme immunoassay and immunohistochemistry as standards, the overall accuracy for ER was 97% and for PR was 89%. The results of this study show that the ThinPrep smear with microwave antigen retrieval pretreatment is a reliable method and a suitable alternative for hormone receptor analysis in breast carcinoma.     

Immunohistochemical study of hormone receptor and hormone-regulated protein expression in phyllodes tumour: comparison with fibroadenoma

 The histogenesis of phyllodes tumour (PT) and that of fibroadenoma (FA) of the breast appear to be closely related. FA is thought to be hormonally responsive, while the hormone-responsiveness of PT is uncertain. To gain insight into hormone-responsiveness of PT, we performed immunohistochemical analysis of oestrogen-regulated pS2 and androgen-regulated prostate-specific antigen (PSA) protein expression and also of oestrogen receptor (ER), progesterone receptor (PgR) and androgen receptor (AR) expression in paraffin sections obtained from 50 female PT patients. Paraffin sections taken from 50 female fibroadenoma (FA) patients were analysed for comparison. ER, PgR, pS2, AR and PSA expression were detected in 32%, 96%, 20% 98% and 4.0% of PT sections and in 28%, 96%, 42% 80% and 10% of FA sections, respectively. No correlations were detected among ER, PgR and pS2 expression or between AR and PSA expression in PT or FA sections. PgR expression was significantly associated with AR expression in PT (P<0.0001). The present investigations indicate that PT and FA have almost similar hormone receptor status. However, different positivities of pS2 expression suggest that oestrogen-responsiveness may differ between PT and FA. In addition, a wide-ranging co-expression of AR and PgR in PT sections suggests that these receptors may play an important part in the proliferation, although the functional significance of these receptors should be elucidated. Received: 26 January 1998 / Accepted: 5 April 1998     

Comparative analysis of various biochemical and immunohistochemical methods for evaluating hormone receptors in breast carcinoma

The aim of this study was to compare the grade of concordance between three methods to evaluate and to quantify the expression of estrogen and progesterone receptors in 180 cases of breast cancer. The methods compared were dextran-coated charcoal (DCC) method, immunochemical analysis (ICA), and immunohistochemistry (IHC). The grades of concordance are so summarized: (1) 77.8% for estrogen receptors (ER) and progesterone receptors (PgR) with DCC and ICA methods (r = 0.53 and r = 0.64, respectively); (2) 83.3% for ER (r = 0.53) and 75.6% for PgR (r = 0.64) with DCC and IHC methods; (3) 87.8% for ER (r = 0.98) and 86.7% for PgR (r = 0.99) with ICA and IHC methods. Nowadays, the technique carried out on tissue sections provides the most reliable information. The IHC method performed on formalin-fixed and paraffin-embedded tissue is preferred in laboratories with a high work load because it is easy and inexpensive and can be automated.     

New monoclonal antibodies to oestrogen and progesterone receptors effective for paraffin section immunohistochemistry

Assessment of oestrogen and progesterone receptors (ER and PgR) in breast cancer is widely used for the prediction of response to endocrine therapy and as a prognostic marker. Cytosolic assays have been replaced in many centres by immunochemical techniques, which have many advantages including applicability to small samples, simplicity, and cost-effectiveness. This study describes the generation and characterisation of two novel murine monoclonal antibodies recognizing ER and PgR, designated NCL-ER-6F11 and NCL-PGR respectively, which are effective in heat-treated formalin-fixed, paraffin-embedded tissue. The antibodies have been characterized by Western blotting and by immunohistochemistry on normal and pathological breast and other tissues. NCL-ER-6F11 has been shown to compare favourably with a currently available ER antibody. These antibodies may prove of value in the assessment of hormone receptor status in human breast cancer. © 1997 John Wiley & Sons, Ltd.     

Use of monoclonal antibody for assessment of estrogen receptor content in fine-needle aspiration biopsy specimen from patients with breast cancer

With increasing emphasis on application of fine-needle aspiration biopsy (FNAB) for an earlier detection of breast cancer, there is clearly a need for a method that requires only a small number of tumor cells for hormone receptor analysis. An immunocytochemical assay utilizing monoclonal antiestrophilin antibody and the peroxidase-antiperoxidase technique has been shown to be highly specific and sensitive for the detection of estrogen receptor (ER) in breast cancer tissues. To assess the usefulness of this technique in FNAB, 62 cases of primary, recurrent, and metastatic breast cancers were studied. The findings from the aspirated cells employing estrogen receptor immunocytochemical assay (ER-ICA) were compared with results obtained from cell population of the same tumors following their removal using both cytochemical (ER-ICA) and biochemical (dextran-coated charcoal assay) methods for ER determination. Overall, there was a 92% concordance between biochemical and cytochemical results. This result suggests that anti-ER monoclonal antibody in immunocytochemical analysis is an effective tool in assessment of ER content in breast cancers. The technique can be easily performed at community hospitals and is well suited for specimens of insufficient size for biochemical assay. It may extend the scope of FNAB and make ER studies possible on material unsuitable for biochemical assay from sites other than breast.     

Immunohistochemical detection of the estrogen receptor‐α and progesterone receptor in the canine pregnant uterus and placental labyrinth

The presence of hormone receptors is as important as the amount of hormone to predict hormone action. Therefore, the presence of estrogen receptors of the alpha subtype (ER‐α) and progesterone receptors (PR) was evaluated in six pregnant uteri including the placenta and in three postpartum uteri of dogs. This preliminary study is part of our immunohistochemical research project on steroid hormone receptor distribution in the canine female genital tract. Specific staining for ER‐α or PR was found only in cell nuclei. Staining for ER‐α was rare in the various cell types of pregnant and postpartum uteri. Staining for PR was absent or weak in epithelial cells. Moderate staining for PR was observed in endometrial stromal cells and myometrial smooth muscle cells, two cell types playing an important role in the maintenance of pregnancy. Stromal cells stained more frequently positive for ER‐α and PR than epithelial cells, indicating that both hormones may act on epithelial cells indirectly via stromal cells. In the placental labyrinth, fetal cells showed no evidence of ER‐α or PR. In contrast, both receptors were present in maternal mesenchymal cells that were located around the basement membrane of the maternal blood vessels. These cells showed signs of decidualization. No difference in PR distribution was seen between pregnant and postpartum uterine tissue, suggesting that during parturition the decrease in serum progesterone levels and the concomitant increase in the estrogen/progesterone ratio are probably more important than the decline in receptor availability. Anat Rec 260:42–50, 2000. © 2000 Wiley‐Liss, Inc.     

Discrepancies of the biochemical and immunohistochemical estrogen receptor assays in breast cancer

We examined the estrogen receptor (ER) content of 124 primary breast cancers by hormone binding and immunohistochemical (ER-ICA) assays. Both assays were in agreement in 110 tumors (89%; P less than .0001); 68 tumors were positive and 42 were negative. In 14 cases (11%), the assays yielded discordant results. Three tumors showed hormone binding in the absence of immunohistochemically detectable ER; the false positive hormone binding resulted from the presence of normal epithelium adjacent to ER-ICA negative malignant cells. Eleven tumors failed to show hormone binding but were ER-ICA positive. Four of these were from premenopausal patients whose circulating endogenous estrogen may occupy the receptor, giving rise to false negative hormone binding assays. In four cases, the discrepancy of negative hormone binding assay and positive ER-ICA assay was attributed to scant tumor cells in the tissue sample. The remaining three discrepancies could not be resolved with certainty, but possibly resulted from alteration of the hormone binding site with preservation of the immunoreactive epitope on the ER molecule. These results indicate that the ER-ICA assay is more accurate than the hormone binding assay in identifying the presence of ER in cancer cells. The heterogeneous immunostaining of ER in tumor sections, which may reflect mosaicism of tumor cells, rate of cell proliferation, or phase of cell cycle, remains unexplained.     

Fatty acid synthase expression in Japanese breast carcinoma patients.

Fatty acid synthase (FAS) is the key enzyme required for the conversion of dietary carbohydrates to fatty acids. Recent studies have demonstrated that high levels of FAS expression occur in a variety of cancers, including breast cancer. We evaluated 243 primary breast cancer patients in the period between 1989 and 1996. Immunohistochemical staining for FAS was performed on formaline-fixed, paraffin-embedded sections. FAS staining intensity was graded as low or high. The expression of FAS was high in 145 (60%) and low in 98 cases (40%). A weak correlation between FAS expression and nodal status was noted in premenopausal patients (p=0. 01). FAS was associated with estrogen receptor (p=0.0022) and progesterone receptor (p=0.0085) status. We found that a low expression of FAS was significantly related to a shorter disease-free survival (DFS) rate in estrogen receptor positive patients (p=0.024) and a similar trend was recognized in progesterone receptor positive patients (p=0.083). The low FAS group showed better DFS and OS in all but ER-/PgR- cases (p=0.011, 0.076). This study showed close correlations between immunohistochemical FAS expression and steroid hormone receptors in premenopausal patients. The use of FAS expression may increase the diagnostic utility of ER and PgR in premenopausal patients. FAS may be able to predict the responsiveness of tumors to endocrine therapy.     

Significance of hormone receptor status and tumor vessels in normal, hyperplastic and neoplastic endometrium

The aims of this study were to identity the roles of tumor vessels and hormone receptor status in normal, hyperplastic, and neoplastic endometrium, and to explore their relationships with other prognostic factors of endometrial adenocarcinoma. Endometrial curettage specimens of proliferative phase and secretory phase endometrium, simple hyperplasia with or without atypia, complex hyperplasia with or without atypia, and grade 1 adenocarcinoma were examined for estrogen receptor alpha (ER alpha), progesterone receptor (PgR), Ki-67 labeling index (LI), cyclin D1, microvessel density (MVD), and area of venules (AV) using an immunoperoxidase method. The results showed high levels of ER alpha in complex hyperplasia, and high levels of PgR in simple hyperplasia without atypia. Expression of ER alpha in the endometrium decreased in a stepwise manner from complex hyperplasia without atypia to grade 1 adenocarcinoma. Expression of PgR in the endometrium decreased in a stepwise manner from simple hyperplasia without atypia to grade 1 adenocarcinoma. In contrast, the expressions of Ki-67 LI, cyclin D1, MVD and AV in the endometrium increased in a stepwise manner from normal, simple or complex hyperplasia with or without atypia to grade 1 adenocarcinoma. These changes may become irreversible on progression from simple or complex hyperplasia to neoplasia.     

Estrogen receptor is significantly associated with the epithelioid variants of renal angiomyolipoma: a clinicopathological and immunohistochemical study of 67 cases

Perivascular epithelioid cells (PEC) in angiomyolipoma (AML) were recently proposed to be its most common progenitor cells. Histologically, triphasic components were present in various proportions, but were overwhelmingly myogenic in epithelioid variants of AML. Despite histological discrimination, the immunophenotypic profiles between triphasic and epithelioid AML have never been compared. The aim of the present study was to clarify the identity of PEC by using immunoreactivity to estrogen receptor (ER), progesterone receptor (PR), bcl-2 and placenta alkaline phosphatase (PLAP), and to use this information to compare triphasic and epithelioid AML. A total of 33 out of 67 cases of renal angiomyolipoma that underwent surgery were reviewed over the period 1998-2003. Two cases were associated with tuberous sclerosis. Ten patients had other malignant tumors, and three patients had a nodal extension. Immunohistochemistry showed that bcl-2 (59.4%), PLAP (46.9%), HMB-45 (100%) was predominantly localized around vessels. The stem cell markers were absolutely negative in all AML types. The estrogen receptors were positive in 14 cases (42.4%) and the progesterone receptors were positive in five cases. Bcl-2 and both female sex hormone receptors were significantly more frequent in the epithelioid variant of AML than in the triphasic type. Perivascular epithelioid cells express bcl-2, ER, PR and PLAP, and ER could be partly associated with myogenic proliferation.