Papillary neoplasia of the breast: immunohistochemically defined myoepithelial cells in the diagnosis of benign and malignant papillary breast neoplasms

The presence or absence of myoepithelial cells (ME) has been considered as an important feature in the differential diagnosis of benign and malignant papillary lesions of the breast. We evaluated the distribution of myoepithelial cells in formalin-fixed paraffin-embedded tissue sections of 25 papillomas and 18 papillary carcinomas by ABC immunoperoxidase technique with antibodies to muscle actin (HHF-35) and high molecular weight (HMW) keratin (clone 34BE12, cytokeratins 1, 5, 10, and 14; reacting preferentially with ME cells) and an antiserum to S-100 protein. Also included in the study were eight cases of micropapillary ductal carcinoma in situ (DCIS) having a few fibrovascular cores and five peripheral papillomas with accompanying ductal carcinoma in situ or atypical hyperplasia. The antibodies to muscle actin were sensitive and relatively specific for ME cells of the breast and uniformly labeled ME cells in all 25 papillomas. ME cells were absent or extremely sparse in papillary carcinomas. They were present focally in some of the fibrovascular cores of the micropapillary DCIS, and a mixed pattern was observed in peripheral papillomas with areas of carcinoma. HMW keratin was variably expressed in ME cells in most cases with positive internal controls and was present in several normal ductal and papilloma epithelial cells but not in epithelial cells of papillary carcinomas. HMW keratin, although less specific for ME cells, was a useful adjunct because of its reactivity with ME cells as well as hyperplastic epithelial cells in papillomas, which resulted in a combined positive reaction.(ABSTRACT TRUNCATED AT 250 WORDS)     

Validation of immunolocalization of the urokinase receptor expression in ductal carcinoma in situ of the breast: comparison with detection by non-isotopic in-situ hybridization

AIMS: Ductal carcinoma in situ (DCIS) is a pre-invasive form of mammary carcinoma with no microscopic evidence of cancer cell invasion through the basement membrane. However, for initiation of invasion, tumour cells have to acquire and focus proteolytic activity on to the cell surface in order to infiltrate the surrounding extracellular matrix. The receptor (uPA-R or CD87) for the serine protease urokinase-type plasminogen activator (uPA) plays a central role in invasion and metastasis. This study was performed to determine and localize m-RNA and protein of uPA-R in ductal carcinoma in situ of the breast. METHODS AND RESULTS: We analysed uPA-R mRNA and protein expression by in-situ hybridization and immunohistochemistry, respectively, in 50 formalin-fixed, paraffin-embedded specimens of DCIS. Three different antibodies were used to stain cell-associated uPA-R; chicken polyclonal antibody (pAb) HU277 and monoclonal antibodies (mAb) IID7 and 3936. In all cases, myoepithelial and stromal cells reacted with either antibody. Especially, reaction of macrophage-like cells with mAb 3936 resulted in a well-marked and bright staining. Applying mAb IID7, in 46 of the 50 breast specimens tumour cells showed a positive immunoreaction. Likewise pAb HU277 stained tumour cells in 40 of the 50 cases, whereas mAb 3936 reacted with only 24 of the 50 tissue sections. Endothelial cells were marked by both mAb IID7 and pAb HU277 (46/50 and 35/50, respectively); mAb 3936 did not label at all. All of the cell types stained by mAb IID7 and pAb HU277 also displayed reactivity with uPA-R mRNA-specific antisense oligonucleotides in in-situ hybridization. CONCLUSIONS: Our results reveal the presence of the tumour invasion-related receptor for the protease uPA not only in invasive ductal breast carcinoma but also in different types of DCIS.     

Involucrin expression in breast carcinomas: An immunohistochemical study

The expression of involucrin, a structural component of the envelope of mature squamous epithelium, was studied in 166 paraffin-embedded breast carcinomas. In 41 cases (24.7%) involucrin-positive, light microscopically non squamous tumour cells were detected. The number of involucrin-positive tumour cells varied considerably from case to case. For further characterization, involucrin-positive cases were studied using monoclonal antibodies to various cytokeratins (PKK1, EAB 903, EAB 904) and, in selected cases, double immunostaining with antibodies to cytokeratins and involucrin were performed. Coexpression of involucrin and cytokeratins demonstrated by PKK1 was seen in all tumour cells, whereas coexpression of involucrin and cytokeratins detected by EAB 904 was only seen in single and scattered cells in a few cases. Cytokeratins detected by EAB 903 were not coexpressed with involucrin in our cases. Our results indicate heterogeneity of cytokeratins in breast carcinomas and suggest a dissociation in the regulation of involucrin and cytokeratin expression.     

伴有神经内分泌分化的乳腺梭形细胞癌

目的探讨乳腺伴有神经内分泌分化的梭形细胞癌的病理形态学和免疫表型特点及鉴别诊断。方法复习2500例乳腺癌切片,找出以梭形细胞占主要优势（〉80％）的癌5例,其中2例梭形细胞型导管内癌和3例梭形细胞型浸润癌。采用HE、阿辛蓝（AB）／PAS和网织染色,以及用癌胚抗原（CEA）、上皮膜抗原（EMA）、细胞角蛋白（CK7、3413E12、AE1／AE3）、神经元特异性烯醇化酶（NSE）、突触素、嗜铬蛋白（cg）A、Lue-7、波形蛋白,S-100、平滑肌肌动蛋白（SMA）、calponin、雌激素受体（ER）、孕激素受体（PR）、c—erbB-2、E-钙黏素、Ki-67、p53抗体进行免疫组织化学观察。其中4例有随访信息。结果患者平均年龄在68岁。镜下：5例癌细胞形态主要为长梭形的上皮样细胞,3例有少数胞质内空泡状细胞,4例可见散在AB阳性细胞。免疫组织化学5例均表达AE1／AE3、EMA、CEA、E-钙黏素和突触素,CK7有4例表达,NSE阳性3例,CgA和Lue7阳性2例,ER阳性4例,PR阳性2例,1例表达c-erbB-2,1例有灶状波形蛋白阳性。免疫组织化学结果显示2例梭形细胞型导管内癌和1例梭形细胞型浸润性癌是梭形细胞型的神经内分泌癌,另外2例梭形细胞型浸润性癌是伴有神经内分泌分化的化生性癌。随访3例存活（24～58个月）,1例27个月内死亡。结论上皮样梭形细胞和细胞内黏液的出现是乳腺伴有神经内分泌分化癌的一个形态学特点。梭形细胞神经内分泌型导管内癌需要和普通导管增生及导管内乳头状瘤鉴别。梭形细胞型的神经内分泌癌和伴神经内分泌分化的梭形细胞浸润性癌需要与梭形细胞肌上皮肿瘤、恶性黑色素瘤及某些软组织肿瘤鉴别。     

The nature of breast dense core granules: chromogranin reactivity

Ultrastructural immunoreactivity for chromogranin with the LK2H10 antibody was examined in nine cases of breast carcinoma and one example of normal resting breast. These tissues were selected for study on the basis of argyrophilia or LK2H10 immunostaining by light microscopy. In two cases, positive reactivity with the chromogranin antibody was seen in breast neuroendocrine-like dense core granules. In a further six cases chromogranin reactivity was not seen in similar granules that showed the neuroendocrine properties of ultrastructural argyrophilia and uranaffinity. Inability to exhibit the full complement of neuroendocrine characteristics in breast carcinomas contrasts with the facility to demonstrate them in tissues of usual neuroendocrine differentiation. Furthermore, in two mucoid carcinomas and one example of normal resting breast ultrastructural cytoplasmic LK2H10 reactivity was evident, which was not localized to dense core granules, although these were present in two of the cases. Our findings demonstrate the heterogeneity of breast dense core granules and discourage acceptance of such characteristics as evidence of histogenesis of these carcinomas from a neuroendocrine cell type in breast tissue.     

There is more than one kind of myofibroblast: analysis of CD34 expression in benign,in situ,and invasive breast lesions

AIMS: Smooth muscle actin (SMA) positive myofibroblasts have been implicated in tumour invasion; however, acquisition of SMA is not limited to peritumorous fibroblasts and other changes in fibroblasts may be more specifically related to the malignant environment. CD34 is a sialomucin expressed by normal breast fibroblasts but lost in invasive carcinomas. The aim of this study was to establish the relation between CD34 and SMA expression in breast fibroblasts and to analyse whether loss of CD34 is specific for invasive disease. METHODS: Immunohistochemistry for CD34 and SMA was performed on 135 cases including 10 normal, 10 fibroadenomas, 40 infiltrating ductal carcinomas, 55 cases of ductal carcinoma in situ (DCIS), and 20 radial scar/complex sclerosing lesions. The relation between staining pattern and histopathological features was recorded as positive, negative, or reduced. RESULTS: Fibroblasts around all normal duct-lobule units and those showing epithelial hyperplasia were CD34 positive and mainly SMA negative. In fibroadenomas, fibroblasts retained CD34 but acquired SMA expression. In contrast, fibroblasts around invasive carcinoma were CD34 positive and SMA negative. In DCIS, loss of CD34 was significantly more frequent in high grade tumours than in low or intermediate grade ones (p < 0.001). The acquisition of SMA was seen more frequently than the loss of CD34, particularly in non-high grade DCIS. In all radial scars, fibroblasts were SMA positive but CD34 negative, and a similar pattern was seen in stromal cells in areas of fibrosis following core biopsy. CONCLUSIONS: These results show that SMA positive myofibroblasts exhibit variable expression of CD34, indicating that these markers are not coordinately controlled. Loss of CD34 is strongly related to the malignant phenotype, in both invasive and preinvasive disease, but is not entirely specific because radial scar fibroblasts and fibroblasts in reactive fibrosis exhibit a similar phenotype. The functional relevance of altered CD34 expression is unclear but the very focal changes implicate local signalling mechanisms probably of epithelial origin.     

乳腺神经内分泌型导管内癌

目的对乳腺神经内分泌型导管内癌（E-DCIS）的临床病理特点、预后和鉴别诊断进行探讨。方法用光镜、免疫组织化学EnVision法行嗜铬素A、突触素和神经元特异性烯醇化酶（NSE）染色和消化PAS、消化阿辛蓝和嗜银染色,对18例具有E-DCIS特征的乳腺癌进行观察。结果具有E-DCIS特征的乳腺癌具有以下特点：（1）好发于老年女性,平均年龄71岁。最常见的临床症状为乳腺肿块或乳头溢液。（2）E-DCIS呈导管内肿瘤细胞的膨胀性生长,在肿瘤周边常可见导管内乳头状瘤。（3）肿瘤细胞呈多边形,卵圆形或梭形,胞质丰富,嗜酸性或细颗粒状。细胞核往往只有轻一中度异型,消化PAS或AB染色显示细胞内或细胞外存在黏液,有些肿瘤细胞呈印戒细胞样。（4）〉50％的肿瘤细胞表达嗜铬素A、突触素和NSE中的至少两种,部分病例CD56和CD57染色阳性。（5）E-DCIS中常可见到肿瘤细胞向邻近导管的派杰样扩散,且在膨胀性生长的导管内不存在肌上皮成分。这两点有助于E-DCIS与导管上皮增生的鉴别。结论E-DCIS是一种低度恶性的乳腺导管内癌,有其独特的组织形态、免疫组织化学特征,应作为一种独立的导管内癌类型加以认识。     

A morphological and immunocytochemical study of a distinctive variant of ductal carcinoma in-situ of the breast

Because so-called 'carcinoid' tumour of the breast has proven to be a difficult entity to define, we studied in-situ carcinoma as there were reasons to believe that this might help clarify the complex problems involved. We studied a consecutive series of 30 cases of ductal carcinoma in-situ (DCIS) by light microscopy and silver impregnation methods and identified a relatively common endocrine variant of DCIS. This variant was studied by immunocytochemical and ultrastructural methods, using conventional DCIS as a control. Endocrine DCIS is identified by its organoid pattern, stromal 'inclusions', festooned structure and a distinctive type of polypoid invagination. It is argyrophilic and rich in neuron-specific enolase. Ultrastructurally it contains abundant dense core granules which are impregnated selectively by Grimelius' method. This tumour type frequently contains peptide hormones of the ACTH family. Three of seven cases contained cells reactive for ACTH and corticotropin-like intermediate lobe peptide CLIP or their precursor, pro-opiomelanocortin. A fourth tumour contained neurotensin, recently identified in a variety of endocrine tumours. Argyrophil invasive carcinomas are a much more heterogeneous group of tumours than argyrophil DCIS and only a minority have an endocrine structure comparable to that described here.     

Expression of steroid hormone receptors, their regulated proteins, and bcl-2 protein in myofibroblastoma of the breast

AIMS: To investigate the possible role of steroid hormones in the pathogenesis of myofibroblastoma (MFB) of the breast, we analysed the immunohistochemical expression of oestrogen, progesterone, androgen receptors, their regulated proteins and bcl-2 protein in a series of this rare tumour. METHODS AND RESULTS: Paraffin-embedded sections from seven cases of MFB of the breast (five male; two female) were immunohistochemically tested for the expression of oestrogen receptor (ER), progesterone receptor (PR), androgen receptor (AR), oestrogen-regulated pS2 protein, androgen-regulated prostate-specific antigen (PSA) and bcl-2 protein. Rare cases of benign spindle cell tumours or tumour-like lesions of the breast (primitive fibromatosis, inflammatory pseudotumour, muscular hamartoma) which enter into the differential diagnosis with MFB, were also investigated and compared with MFB. All cases of MFB showed a diffuse (70-90% of neoplastic cells) and strong nuclear labelling with ER and PR, whereas AR was expressed only in three cases (two men and one woman) in about 60-70% of cells. Conversely, no immunostaining was detected for the pS2 protein and PSA. bcl-2 protein immunoreactivity was found in all cases of MFB, although with a variable degree of expression. No expression for steroid hormone receptors, their regulated-proteins and bcl-2 protein was observed in the rare benign spindle cell lesions of the breast included in this study. CONCLUSION: The in-situ detection of ER, PR and AR suggests that steroid hormones and their receptors are implicated in the pathogenesis of breast MFB. The consistent demonstration of bcl-2 protein, associated with a positive ER/PR status, provides evidence that bcl-2 may be an oestrogen-regulated protein also in MFB and that probably plays a role in the tumorigenesis. Finally, we postulate that the ER/PR and bcl-2 positive immunoprofile of MFB of the breast, in contrast to the negative profile of other rare primitive benign spindle cell lesions of the breast herein studied, might be exploited as an ancillary diagnostic aid in differential diagnosis of doubtful cases.     

Rhabdoid tumour of the lung is a dedifferentiated phenotype of pulmonary adenocarcinoma

AIMS: Primary rhabdoid tumour of the lung is rare, and histological and biological characteristics have not been fully documented. We describe three cases of primary lung rhabdoid tumour, all associated with adenocarcinoma, and investigate the histological features and biological characteristics. METHODS AND RESULTS: Three cases were obtained from a total 902 cases of surgically removed primary lung tumours between 1986 and 1998. The rhabdoid cells were found to occupy about 50-90% of each tumour. All of the tumours had nonrhabdoid adenocarcinoma foci in the centre of the tumours. Transition between the adenocarcinomatous and rhabdoid components was demonstrated. Detailed immunohistochemical studies were carried out. The epithelial markers, cytokeratins and epithelial membrane antigen (EMA), were strongly expressed in rhabdoid and adenocarcinomatous components. Furthermore, surfactant apoprotein A was positive in both components in one case, but myoglobin, MyoD and HHF35 were not expressed. Vimentin was strongly and diffusely stained in all cases. The neuroendocrine markers, chromogranin A (all cases), neuron-specific antigen (NSE) (two cases) and CD56 (one case) were occasionally positive in only a small number of the rhabdoid tumour cells. GM-CSF was positively stained in one case, and the dedifferentiated characteristics of the rhabdoid cells was suggested. Proliferative cell nuclear antigen (PCNA) was strongly demonstrated in the rhabdoid tumour cells (all cases). To gain better understanding the highly proliferative characteristics of the tumours, p53 gene (exons 5-8) mutation was examined by DNA sequencing analysis; mutation of the p53 DNA was not detected. Overexpression of p53 protein was also not demonstrated in all cases. HPV6 was demonstrated in one case by PCR method and also non-isotopic in-situ hybridization (NISH). Two cases died in a short period of time (3 years and 4 months, respectively). CONCLUSION: The rhabdoid cells in these three cases were considered to represent the dedifferentiated components of the accompanying adenocarcinoma. Dedifferentiated characteristics (neuroendocrine markers, GM-CSF, vimentin, and the aggressive behaviour) were evident.     

Value of cytokeratin 5/6 immunostaining using D5/16 B4 antibody in the spectrum of proliferative intraepithelial lesions of the breast. A comparative study with 34βE12 antibody

Previous studies have shown that basal-type cytokeratins (CKs) can distinguish usual ductal hyperplasia (UDH) from the spectrum of atypical ductal hyperplasia (ADH), ductal carcinoma in situ (DCIS) and lobular carcinoma in situ (LCIS). Indeed, expression of these CKs is weak or absent in ADH, DCIS and LCIS. However, the diagnostic usefulness of D5/16B4 antibody (anti-CK5/6) has never been compared with that of 34betaE12 antibody (anti-CK1/5/10/14). We performed immunostaining of CK 5/6 and CK1/5/10/14 on 100 breast lesions, including UDH ( n=31), ADH ( n=5), DCIS ( n=54) and LCIS ( n=10). Abundant immunostaining was observed in all UDH using both antibodies. Four of five of the ADH cases showed less than 5% of CK5/6 stained cells, the remaining case showed 30% of labeled cells. With 34betaE12 antibody, three of five of the ADH cases showed less than 5% labeled cells, while two cases showed more than 30% of stained cells. None of the 54 DCIS or the 10 LCIS was labeled by D5/16B4, while a lack of 34betaE12 immunostaining was observed in only 15 of 54 DCIS and 2 of 10 LCIS. We confirmed that D5/16B4 antibody directed against CK5/6 is useful in distinguishing UDH from the spectrum of ADH/DCIS/LCIS. We also demonstrated that D5/16B4 is far a more specific marker than 34betaE12 antibody.     

p53 mutation in breast cancer. Correlation with cell kinetics and cell of origin

AIM: Several studies have investigated the expression of the cytokeratins (CKs), vimentin, the epithelial growth factor receptor (EGFR), the oestrogen receptor (ER), and the progesterone receptor (PgR), in breast cancer, but no study has directly compared p53 mutations with these phenotypic and differentiation markers in the same case. The present study was designed to provide some of this information. METHODS: The expression of the p53 and bcl-2 proteins was evaluated by immunohistochemistry in relation to phenotypic characteristics and cellular kinetic parameters (mitotic index and apoptotic index) in 37 cases of ductal carcinoma in situ (DCIS) and 27 cases of infiltrating ductal carcinoma (IDC) of the breast. In addition, p53 gene mutation was examined by polymerase chain reaction single strand conformation polymorphism analysis (SSCP). RESULTS: Thirteen cases (eight DCIS and five IDC) showed expression of CK8, CK14, CK18, vimentin, and EGFR, consistent with a stem cell phenotype, whereas 44 cases (27 DCIS and 17 IDC) showed expression of CK8 and CK1, weak or negative expression of CK18, but were negative for vimentin and EGFR, consistent with a luminal cell phenotype. DCIS and IDC cases with a stem cell phenotype were ER/PgR negative and intermediately or poorly differentiated. In contrast, the cases with luminal cell phenotype were ER/PgR positive and well or intermediately differentiated. In addition, intermediately or poorly differentiated cases with a stem cell phenotype showed higher proliferative activity (per cent of MIB-l positive cells) than did intermediately or well differentiated cases with a luminal cell phenotype. Both DCIS and IDC cases with a stem cell phenotype were p53 positive and bcl-2 negative by immunohistochemistry. In IDC, p53 expression was associated with a reduction of both mitotic index and apoptotic index compared with DCIS. Most of the tumours showing a more differentiated phenotype (luminal) were p53 negative and bcl-2 positive. In these cases, cell kinetic parameters increased from DCIS to IDC. These data suggest the existence of subsets of DCIS and IDC that, because of their phenotypic characteristics, could be derived from subpopulations of normal breast cells having different control mechanisms of cell proliferation and neoplastic progression. CONCLUSIONS: These results are compatible with the hypothesis that the phenotype of the cell of origin constrains both tumour phenotype and the choice of genetic events; however, the occurrence of p53 mutants by chance during neoplastic transformation cannot be excluded.     

Distribution of calponin and smooth muscle myosin heavy chain in fine-needle aspiration biopsies of the breast

The cell types that may be present in any fine-needle aspiration biopsy (FNAB) of breast include epithelial cells (EC), myoepithelial cells (MEC), bipolar stromal cells (BSC), vascular pericytes/endothelial cells (VPEC), and adipose cells (AC). The recognition of most of these benign cellular elements in aspirates of the breast is relatively straightforward, based on distinct cytomorphologic criteria. However, there is controversy regarding the recognition of MEC because BSC are often referred to as MEC by cytopathologists. It is important to identify MEC in breast aspirates, because their presence has been associated with benign epithelial proliferations. In this study we used immunocytochemical methods on archival cytology slides with antibodies specific for MEC, calponin, and smooth muscle myosin heavy chain (SMMHC), to determine the distribution of MEC in FNAB of the breast and to ascertain the distribution of MEC in in situ and invasive carcinomas. Fifteen benign FNABS of breast and corresponding tissue biopsies were obtained along with 10 malignant FNABS and corresponding excisional breast biopsies from 1989-1993. Calponin and SMMHC antibodies were used on archival alcohol-fixed Papanicolaou-stained direct smears as well as the corresponding tissue sections. The distribution and pattern of positive immunostaining with both antibodies were recorded on the benign elements and the carcinomas for both cytologic and histologic slides. Benign breast tissues demonstrated strong continuous immunostaining for calponin and SMMHC of MEC. The interlobular stromal cells as well as intralobular stromal cells showed no immunostaining with either antibody. In cytologic preparations, MEC staining with calponin and SMMHC appeared as spindle cells between epithelial cells or along the edges of the epithelial groups. The bipolar stromal cells did not stain with either antibody. The tissues with DCIS (ductal carcinoma in situ) often showed the presence of MEC with strong calponin immunostaining, but sometimes the immunostaining was discontinuous or entirely absent around markedly dilated ducts. The SMMHC antibody was invariably negative, with architectural DCIS in dilated ducts. Two cases of DCIS with prominent periductal fibrosis or inflammation were positive for calponin, but the periductal stromal cells were calponin- and SMMHC-negative. Invasive carcinoma was negative for both calponin and SMMHC, but areas of DCIS were often positive in a discontinuous pattern. In conclusion, 1) Benign cellular elements from breast tissue FNAB showed strong continuous decoration of MEC with both calponin and SMMHC. Vascular pericytes and vascular smooth muscle were positive for both antibodies, but these cells were not observed in the FNAB. Benign proliferative epithelium showed no local increase in MEC with either antibody. Bipolar stromal cells in tissue and smears did not stain for MEC antibodies. 2) BSC did not correspond morphologically to MEC, and were not decorated with calponin or SMMHC. 3) Calponin-positive MEC were commonly associated with in situ ductal lesions, although they may at times have been discontinuous or absent entirely. DCIS may be recognized in FNAB by the presence of calponin-positive MEC associated with tumor cell groups. 4) Invasive carcinomas were invariably negative for MEC with these antibodies.     

Expression of phosphorylated and non-phosphorylated neurofilament subunits and cytokeratins in neuroendocrine lung tumors

In this study antibodies specific for different intermediate-sized proteins (cytokeratins and neurofilaments) have been tested on a series of neuroendocrine (NE) lung tumors in order to evaluate their diagnostic validity. In particular we used a panel of polyclonal anti-neurofilament 200-kilodalton subunits whose reactivity against phospho-dependent epitopes was known. At least one NF subunit was constantly present and in all cases coexpression of cytokeratins and neurofilaments was confirmed. However, in cases of carcinoid tumor (CT) the results were homogeneous, while the cases of small cell lung carcinoma (SCLC) showed a much wider range of immunostaining. Our investigation confirms the hypothesis that the phosphorylation state is a significant determinant of immunohistochemical properties of neurofilaments. This might explain the large number of negative results obtained in previous investigations on NE tumors. The phosphorylation of neurofilaments may also be considered an indication of the degree of differentiation of the tumor.     

Neuroendocrine differentiation and prognosis in breast adenocarcinoma

AIMS: Neuroendocrine differentiation has been detected, and its prognostic value studied, in a number of common human carcinomas. To date there are few detailed studies examining its relevance in breast carcinoma. In this study we evaluate the frequency and prognostic importance of neuroendocrine differentiation in breast adenocarcinoma. METHODS AND RESULTS: The presence of neuroendocrine differentiation, defined as positive reactivity for three markers, neuron-specific enolase (NSE), chromogranin A and/or synaptophysin, has been evaluated in 99 patients with primary operable breast cancer using standard immunocytochemical techniques. A consecutive cohort of patients were selected from the Nottingham/Tenovus series. Comprehensive patient and tumour records have been maintained, and patients were followed up according to a defined protocol. Eighteen cases were positive for NSE, 10 for chromogranin A and 13 for synaptophysin. Eleven percent were positive with more than one neuroendocrine marker. No significant association was found between neuroendocrine differentiation and tumour size, grade, stage or the prevalence of vascular invasion. There was no significant difference in either overall or disease-free survival between patients with or without neuroendocrine differentiation. CONCLUSIONS: In this study we confirm that neuroendocrine differentiation can be identified in a subset (10-18%) of human breast carcinomas. This phenomenon appears to have no relationship to established prognostic factors or patient outcome.     

Cytokeratin 5/6 immunohistochemistry assists the differential diagnosis of atypical proliferations of the breast

AIMS: This study was performed to determine the diagnostic value of keratin 5/6 (CK 5/6) immunophenotyping on routinely processed breast tissues. METHODS AND RESULTS: Six hundred and ninety-nine breast lesions, including normal tissues as well as benign and malignant lesions in 321 formalin-fixed, paraffin-embedded samples from 158 different patients were investigated immunohistochemically, following wet autoclave pre-treatment for antigen retrieval. In normal breast tissues, both myoepithelial and luminal epithelial cells expressed CK 5/6 in varying amounts. While myoepithelial immunoreactivity was most pronounced in the duct system, luminal epithelial immunoreactivity was strongest in the terminal duct lobular units. In ductal hyperplasias (DH), luminal epithelial cells predominantly revealed CK 5/6 immunoreaction. In contrast, neoplastic epithelial cells in atypical ductal and lobular hyperplasias (ADH and ALH) lacked such an expression, whereas in ductal in-situ carcinomas (DCIS) and in infiltrating ductal carcinomas 3.7% and 7.7%, of the cases respectively, showed positive immunostaining for CK 5/6. CONCLUSIONS: Immunophenotyping of keratin 5/6 expression can be helpful in the diagnosis of atypical hyperplasias and in-situ carcinomas of the breast. It is particularly valuable in the differential diagnosis between benign and atypical proliferative lesions.     

Breast carcinoma in women 35 years and younger: a pathological study

AIMS: To document the pathological features of breast carcinoma diagnosed in women aged 35 years or less. METHODS: The files of the Department of Pathology, Singapore General Hospital, were searched for cases of breast cancer diagnosed in individuals aged 35 years or less between January 1993 and December 2004. Histological slides and pathology reports were retrieved and reviewed. Pathologic parameters of tumour size, histological grade, accompanying ductal carcinoma in situ (DCIS), lymphovascular invasion, nodal status, hormone receptor and c-erbB-2 profiles, were documented. RESULTS: Of 112 cases of breast cancer, 91 (81.3%) were invasive carcinomas, 17 (15.2%) pure DCIS, three (2.7%) were diagnosed on needle aspirates. No residual tumour was available for microscopic assessment in one (0.9%) patient who underwent bilateral mastectomy in our institution but had her initial surgery elsewhere. Invasive tumour size ranged from 0.3 to 11.5 cm (mean 2.7 cm, median 2.1 cm), with 84 (92.3%) infiltrative ductal, two (2.2%) lobular, two (2.2%) mucinous, two (2.2%) atypical medullary, and one (1.1%) mixed ductal-lobular. The majority were grade 3 (54 cases, 59.3%), with 24 (26.4%) grade 2 and 7 (7.7%) grade 1, while grading was not accomplished in six (6.6%) cases. Nodal status was positive in 39 (42.9%), negative in 25 (27.4%), unknown in the rest (27 cases, 29.7%). Oestrogen and progesterone receptors (ER, PR) were positive in 51 (61.4%) and 43 (51.8%) cases, respectively, out of 83 (91.2%) cases in which they were evaluated. c-erbB-2 immunostaining, carried out in 54 (59.3%) invasive cancers, showed positivity in 16 (29.6%) cases.DCIS cases ranged from 0.25 to 6.2 cm (mean 2.2 cm, median 2 cm) in size. Nuclear grade was low in seven (41.2%), intermediate in four (23.5%), and high in six (35.3%). CONCLUSION: The majority of breast carcinomas in young women are invasive, with T2 disease at presentation, and of poor histological grade. The recent rise in numbers suggests increased detection, plausibly due to improved awareness of breast disease among the younger female population. Pathogenetic causes that differ from breast carcinogenesis in older women have to be further clarified.     

β1 and β4 integrin expression in methacarn and formalin-fixed material from in situ ductal carcinoma of the breast

The integrins are αβ heterodimeric transmembrane proteins mediating cell-substratum as well as cell-cell interactions. Previous distribution studies on integrin expression have been limited by the requirement for cryostat sectioned tissues, and consequent poor resolution. We have examined 40 examples of ductal carcinoma in situ (DCIS) for the expression of both β1 and β4 integrin chains. These showed strong polarized membrane staining of residual myoepithelial cells (correlating with expression of smooth muscle specific actin) and of the basement membrane region with β1 and β4 antibodies respectively. In 12 out of 40 cases, the DCIS was negative for the β1 chain and a variable pattern of reactivity was seen in the remaining cases. The β4 chain was detected focally and weakly in the tumour cells of 7/40 DCIS and strongly in one; all of these cases were also positive for the β1 chain. Of the 22 cases where co-existent invasion was present, the infiltrating component showed either a similar degree or a diminution of the extent of immunostaining when compared with the in situ component; only one showed enhanced staining (β1 only). This study demonstrates that two of the main β chains, β1 and β4, can be effectively demonstrated on methacarn and cold (4°C) formalin-fixed tissues by avidin-biotin indirect immunoperoxidase staining and that the results are similar to those achieved using frozen tissue.     

Neuroendocrine differentiation in pure type mammary mucinous carcinoma is associated with favorable histologic and immunohistochemical parameters.

Mucinous carcinoma of the breast is a specific good prognostic type malignancy occurring in elderly patients. Neuroendocrine differentiation has long been described in mucinous carcinoma, but the significance of such finding is uncertain. We evaluated the neuroendocrine differentiation profiles of 38 cases of pure mucinous carcinoma and compared the clinicopathological differences between those with and those without neuroendocrine differentiation. The parameters assessed included patients' age, tumor size, nuclear grade, axillary lymph node status at time of diagnosis, percentage area of intratumoral mucin, and the expression of estrogen and progesterone receptors, cerbB2 oncoprotein, and three neuroendocrine markers, namely neurone-specific enolase, chromogranin, and synaptophysin by immunohistochemistry. Patients' outcome and follow-up period were also documented. Of the 38 cases of pure mucinous carcinoma, 28, 11 and six cases showed positive staining for 1, 2 and 3 of the neuroendocrine markers. For all the groups with variable neuroendocrine differentiation and compared to those without such differentiation, they all showed older patients' age, higher proportion of tumors with lower nuclear grade, lower incidence of axillary lymph node metastasis, a higher progesterone receptor, and lower cerbB2 oncoprotein expression. No difference was detected between tumor size, intratumoral mucinous area, and estrogen receptor status. In all, 37 patients did not have distant metastases or local recurrences at the end of follow-up period, while one patient with coexisting high-grade ductal carcinoma in situ at time of diagnosis died of breast carcinoma. Our findings suggest that the identification of neuroendocrine differentiation in pure mucinous carcinoma is associated with more favorable histologic and immunohistochemical parameters.     

乳腺纤维瘤病样梭形细胞癌临床病理分析

目的 探讨乳腺纤维瘤病样梭形细胞癌（fibromatosis-like spindle cell carcinoma,FLSCC）临床病理特征。方法 对3例FLSCC病例进行光镜观察和免疫组化染色[CK、CK（34βE12）、vimentin、SMA、ER、PR、Ki-67、c-erbB-2]。结果 3例均为女性,年龄分别为47、53、56岁,均可触及乳腺肿块。肿瘤境界清楚,但镜下边缘呈浸润性。肿瘤主要是梭形细胞、多边形细胞、少量的管状腺体及鳞上皮巢混合,间质纤维明显增生伴胶原化,细胞成束状排列或散在分布,似纤维瘤病样改变。梭形细胞分化良好,异型性不明显,部分区域细胞较丰富,其间聚集的上皮簇或片状多边形细胞核有轻度异型,可见少数核分裂象。多边形细胞与梭形细胞有移行。病变中亦可见淋巴细胞、浆细胞聚集浸润。上皮细胞、多边形细胞及部分梭形细胞CK（34βE12）、CK（AE1／AE3）阳性,CK阴性的梭形细胞表达vimentin、SMA。3例均行肿块切除,其中1例,术后4个月复发,再行乳腺根治术。结论 乳腺（纤维瘤病样）梭形细胞癌是一种少见的、低度恶性肿瘤,诊断需依赖免疫组化标记并与乳腺其它梭形细胞肿瘤相鉴别。