Immunophenotyping of Mesothelial Cells and Carcinoma Cells With Monoclonal Antibodies to Cytokeratins, Vimentin, Cea and Ema Improves the Cytodiagnosis of Serous Effusions

This paper presents an immunocytochemical study performed on cytocentrifuged deposits from 109 peritoneal and pleural effusions including 20 transudates, 43 malignant metastatic effusions and 46 effusions containing atypical cells, unidentifiable as reactive mesothelial or malignant epithelial cells on the classical morphological criteria. A panel of four monoclonal antibodies (MAb) was used, including KL1 directed to cytokeratins (KER), V9 to vimentin (VIM), NEO 723 to carcinoembryonic antigen (CEA) and E29 to epithelial membrane antigen (EMA). In most transudates the reactive mesothelial cells coexpressed VIM and KER with a ring-like pattern for the latter proteins. In contrast, they were unreactive to anti-CEA and weakly and inconsistently reactive to anti-EMA. In malignant effusions, most carcinoma cells coexpressed EMA, CEA and KER with a predominant diffuse cytoplasmic pattern for the latter. Only a few malignant epithelial cells from five metastatic adenocarcinomas weakly expressed VIM. When used on the 46 effusions with unidentifiable cells, the panel of MAb allowed reactive mesothelial cells and malignant epithelial cells to be distinguished from each other in 39 of 46 cases (85%).     

Characterization of cancer cell lines established from two human metastatic breast cancers

Summary Cell lines are valuable resources for the study of the malignancy and potential therapy of human breast cancer. A major problem with adapting fresh breast tumor specimens to grow in vitro is contamination by fibroblasts. Previously, we have reported a technique to overcome this problem (Nayak, S. K; Dillman, R. O. Clin. Biotechnol. 3:237–242; 1991). We have recently established two new breast cancer cell lines, HH315 and HH375, that were derived from abdominal and supraclavicular lymph node metastases from two patients. They were characterized by (1) growth kinetics; (2) staining with monoclonal antibodies (MoAbs) to cytokeratin-19, epithelial membrane antigen (EMA), anticarcinoembryonic antigen (CEA), breast cancer antigen 1 (BRST-1), breast cancer antigen 2 (BRST-2), Her2/neu, and p53; (3) expression of domains of urinary plasminogen activator (uPA), neural cell adhesion molecule (NCAM), and haptoglobin (Hp) (Harvey et al., 1997); and (4) karyotypic analysis. Growth kinetic studies showed that doubling times for both lines ranged from 48 to 96 h. These two cell lines were found to have characteristics of the metastatic breast cancer cells. Both lines stained positive with MoAbs to cytokeratin-19 and EMA, thus confirming their epithelial origin. They also strongly reacted with the pan-breast carcinoma MoAbs BRST-1 and BRST-2, and carcinoembryonic CEA MoAb. Both cell lines overexpressed the oncogene proteins Her2/neu and p53. The tumor cells were negative for estrogen and progesterone receptors. HH315 cells were poorly differentiated, whereas the HH375 cells exhibited adenocarcinoma morphology. Both cell lines showed intense cell surface and some cytoplasmic staining for uPA, NCAM, and Hp domains, which is a characteristic of malignant neoplasms (Harvey et al., 1997). The HH375 cell line showed two cell types, of which 60% were hyperdiploids with 60–70 chromosomes and 5–10 marker chromosomes. The remaining cells were polyploid with more than 200 chromosomes. Cell line HH315 consisted of only a polyploid population. These cell lines may be useful in breast cancer research.     

CEA, ZGM and EMA localization in cells of pleural and peritoneal effusion: a preliminary study

Carcinoembryonic antigen (CEA), the zinc glycinate marker (ZGM) and epithelial membrane antigen (EMA) have been described as epithelial or tumour markers of varying specificity. These antigens were studied by immunoperoxidase localization in selected cell blocks of 62 pleural or peritoneal effusions and compared to cytological findings and review of the clinical records. By cytological criteria, 25 of the cell blocks were positive for malignancy, 30 negative, and 7 inconclusive. CEA, ZGM, and EMA by immunoperoxidase staining were localized on the cell surface and often in the cytoplasm of malignant cells, in 11/25 (44 per cent), 17/25 (68 per cent) and 22/25 (88 per cent) of the positive cell blocks respectively. Ten (40 per cent) of these cases were positive for all three antigens, 7 (28 per cent) for two, and 6 (24 per cent) for one. Of the 7 cases which were inconclusive on routine cytological reporting, 5 were positive for at least one marker. In 3 of the 5 a diagnosis of malignancy was confirmed, and in the other two was strongly suspected as malignant on clinical grounds. Macrophages were sometimes positive for one or more markers (but showed cytoplasmic staining only) and mesothelial cells in some cases stained positively for EMA but were always negative for CEA and ZGM. Localization of the 3 antigens in cells of malignant effusions was compared with their localization in the primary tumours in 9 cases. Localization corresponded for CEA in 7 of 9 cases, for EMA in 8 of 8 an for ZGM in only 2 of 9. Effusion fluid levels for CEA were compared with the cytological and immunocytochemical findings in 30 cases. Mucin stains performed on the cell blocks were also compared with the immunoperoxidase findings.     

The value of the immunoperoxidase slide assay in the diagnosis of malignant pleural effusions in breast cancer

Whether immunocytochemical studies of malignant pleural effusions due to breast cancer would increase the diagnostic yield as compared with conventional effusion cytology was examined in 30 cases with biopsy-proven metastatic spread to the pleura. Conventional cytology was performed on air-dried smears as well as on cytocentrifuge preparations stained with the May-Grünwald-Giemsa stain. Immunocytochemistry was performed with monoclonal antibodies against carcinoembryonic antigen (CEA), epithelial membrane antigen (EMA) and human leukocyte antigen (HLA) and the peroxidase-antiperoxidase technique on glass slides after Ficoll-Hypaque centrifugation. By conventional cytology, 13 cases (43%) were positive for malignant cells, 6 cases (20%) were suspicious, and 11 cases (37%) were negative. In marked contrast, all 30 cases were immunocytologically positive for malignancy. Tumor cells in all cases demonstrated a positive reaction for EMA. Some mesothelial cells were also positive for EMA, but their reaction pattern was clearly distinguishable from that of the tumor cells. Twenty-one cases (70%) also showed CEA-positive tumor cells; mesothelial cells never reacted with CEA. Some tumor cells showed a loss of HLA expression. In conclusion, this immunocytologic method can be recommended as a routine procedure for greatly increasing the diagnostic yield of cytology in pleural effusions due to breast cancer.     

Identification of carcinoma cells in ascitic and pleural fluid. Comparison of four panepithelial antigens with carcinoembryonic antigen.

The cell membrane antigens epithelial membrane antigen (EMA), epithelial glycoprotein 34, (egp34), BW-495 and tumor-associated antigen 72 (TAG-72) are present in most benign and malignant epithelial cells and can be demonstrated with the help of monoclonal antibodies. In a study on the identification of carcinoma cells in samples of ascitic and pleural fluid involving 170 patients, we compared the value of immunocytochemical labeling of these antigens with that of immunocytochemical demonstration of carcinoembryonic antigen (CEA). Antibodies to EMA and egp34 occasionally also reacted with reactively proliferating mesothelial cells in benign conditions and thus appear to be inappropriate for diagnostic use. Cells positive for BW-495, TAG-72 and CEA, however, have never been found in benign conditions; the specificity of these antigens thus permits their use in diagnosis. Antigen-expressing cells were found in 85% (BW-495), 62% (TAG-72) and 60% (CEA) of cytologically positive samples from carcinoma patients. Similarly, positive reactions for BW-495, TAG-72 and CEA were observed in, respectively, 36%, 29% and 34% of cytologically negative or suspicious samples. BW-495 thus appears to be a suitable marker for the demonstration of carcinoma cells in samples of pleural and ascitic fluid and to have a higher degree of sensitivity than does either TAG-72 or CEA.     

Tumor marker studies of cervical smears. Potential for automation

The distribution of three tumor markers, epithelial membrane antigen (EMA), carcinoembryonic antigen (CEA) and prekeratin (PK) was studied in exfoliated epithelial cells in cervical smears using an immunoalkaline phosphatase staining technique to demonstrate the antigens. EMA was expressed by abnormal cells in a consistent and reproducible fashion whereas the other two markers were variably expressed by both normal and abnormal cells. Our results suggest that immunocytochemical staining for EMA could be of value not only for the diagnosis of cervical intraepithelial neoplasia but also for the automated screening of cervical smears.     

Characterization of the epithelial components in pleomorphic adenoma of the salivary gland

OBJECTIVE: To clarify the cytomorphologic characteristics of luminal epithelial cells (LEC) and neoplastic myoepithelial cells (NMC) in pleomorphic adenoma (PA). STUDY DESIGN: Imprint cytologic smears stained with Papanicolaou stain were examined in 20 patients with PA (including recurrent cases). Immunocytochemistry was performed using the antibodies of epithelial membrane antigen (EMA) and carcinoembryonic antigen (CEA); cells positive for both CEA and EMA were interpreted as LEC and those negative as NMC. RESULTS: LEC were found in 9 of 20 cases as cell clusters in various shapes or as isolated cells with ample cytoplasm. NMC were classified into four types according to their visual patterns and cytoplasmic features: type A, isolated cells with ample cytoplasm; type B, isolated naked cells; type C, cluster of cells with ample cytoplasm; and type D, cluster of cells with scant cytoplasm. NMC were found in all 20 cases, with an absolute incidence of 100%, 90%, 65% and 50%, respectively. CONCLUSION: The different features of NMC (Types A-D) are essential to a specific differential diagnosis. This classification was useful to discriminate PA from other salivary gland tumors with NMC.     

Malignant ascites of serous papillary ovarian adenocarcinoma. An immunocytochemical study of the tumor cells

In 17 malignant peritoneal effusions due to papillary serous adenocarcinoma of the ovary, the reaction patterns of the tumor cells to monoclonal antibodies (MAbs) against surface antigens were studied and compared with the reaction patterns of mesothelial cells in the same effusions. The following surface markers were used with the adhesive slide method: epithelial membrane antigen (EMA), human epithelium-specific cell surface antigen (HEA-125), human endothelial antigen (BMA-120), carcinoembryonic antigen (CEA 3-13), an antibody against natural killer cells and cytotoxic cells (BMA-070), granulocyte antigen (Leu M1) and leukocyte antigen of class I (HLA-1). In all cases, from 30% to 95% of the tumor cells reacted with EMA and HEA-125. Tumor cells showed a positive staining with CEA 3-13 in only five cases. In all cases, from 75% to 95% of the tumor cells reacted positively with BMA-120. The reactivity of a few mesothelial cells with EMA and of all mesothelial cells with BMA-120 did not interfere with the identification of positive tumor cells since the reaction patterns were different. Interestingly, our study demonstrated that BMA-070, an MAb identifying natural killer cells and cytotoxic cells, is also a most useful tumor marker. The same was found to be true for Leu M1, an MAb originally thought to react only with granulocytes. The tumor cells showed a partial or total loss of the expression of HLA-1 reactivity. Since all cases were immunocytochemically positive for tumor cells while conventional cytology was positive in only 13 of the cases, the immunocytochemical analysis of malignant peritoneal effusions due to papillary serous adenocarcinoma of the ovary seems able to improve the cytologic diagnosis of the fluids.     

The value of epithelial membrane antigen in the diagnosis of ovarian tumors.

The value of epithelial membrane antigen (EMA) in the diagnosis of ovarian tumors was investigated using an indirect immunoperoxidase staining technique on 91 histologic sections (88 tumors and 3 normal tissues) and 39 ascitic fluid smears (28 from patients with epithelial ovarian tumors and 11 from cases of myoma uteri). The rate of positive EMA staining was highest in malignant tumors (89.2%), second highest in tumors of low malignant potential (33.3%) and lowest in benign tumors (25.0%); normal ovarian tissues were negative for EMA. Of the malignant tumors, all 48 serous cystadenocarcinomas (100%) and 18 of 26 mucinous cystadenocarcinomas (69.2%) stained positively for EMA. In serous cystadenocarcinomas, the EMA staining was mainly localized on the luminal membrane of cells in well-differentiated tumors, but appeared on the entire cell surface and cytoplasm of cells in poorly differentiated tumors. The results of EMA staining on ascitic fluid smears were almost the same as the results for the histologic sections. The intensity and the localization of EMA staining were related to the grade of malignancy in these ovarian tumors. In comparison with staining for other antigens (carcinoembryonic antigen, CA-125 and human keratin protein), EMA was found to be one of the most sensitive markers for the diagnosis of ovarian cancer.     

Morpho-functional changes in normal cyclic endometrium

Eight cases of normal endometrium in the early proliferative phase and an equal number of cases in the late secretive phase were stained histochemically for acidic mucosubstances and immunocytochemically for epithelial membrane antigen (EMA), carcinoembryogenetic antigen (CEA) and Gross Cyst Disease Fluid Protein -15 (GCDFP-15) production. Alcian blue positive material was observed in both phases of the cycle, although the distribution was different. Sulphated mucosubstances were only present in the secretive phase. No CEA nor GCDFP-15 immunoreactive material was detected. EMA positive staining was observed in all cases, but the distribution varied according to the two different phases of the cycle.     

Distinction between carcinoma cells and mesothelial cells in serous effusions. Usefulness of immunohistochemistry

The usefulness of an immunoperoxidase battery to distinguish carcinomatous from benign effusions was examined. Cell block sections from 90 previously diagnosed effusions were stained with antibodies to Leu-M1, B72.3, epithelial membrane antigen (EMA), carcinoembryonic antigen (CEA) and vimentin. The 90 cases comprised 69 carcinomas (23 mammary, 16 ovarian, 10 pulmonary, 7 gastrointestinal [GI] and 13 others), 2 malignant mesotheliomas and 19 cases with reactive mesothelial cells only. EMA and vimentin were the most useful markers for distinguishing carcinoma cells from reactive mesothelial cells. EMA reacted with 86% of the carcinomas while vimentin reacted with 90% of the reactive cases. Leu-M1, B72.3 and CEA, although generally less sensitive than EMA, were also helpful in this regard. Additionally, the use of Leu-M1 and CEA together may help to distinguish pulmonary from GI carcinomas.     

Immunofluorescent staining of metastatic carcinoma cells in serous fluid with carcinoembryonic antibody, epithelial membrane antibody, AUA-1 and BER-EP4

Using an indirect immunofluorescence technique, we assessed the accuracy and clinical usefulness of a panel of monoclonal and polyclonal antibodies. the panel consisted of carcinoembryonic antibody (CEA) and epithelial membrane antibody (EMA), AUA-1, and Ber-EP4 conjugated with fluorescein isothiocyanate. Twenty-six specimens from pleural, peritoneal or pericardial effusions known to contain carcinoma cells (adenocarcinoma or large cell anaplastic carcinoma) and 16 specimens without carcinoma were first examined. the sensitivity and specificity for each of the antibodies were as follows: CEA, 71% and 75%; EMA, 96% and 81%; AUA-1, 80% and 100%; and Ber-EP4, 85% and 100%, respectively. the panel of antibodies was then applied to a group of 14 'problematic' fluids. These had been identified as causing dilemmas in interpretation, either because the cells in the fluids were of equivocal appearance on light microscopy, or the cytological diagnosis was different from that expected in the light of the clinical condition of the patient. Insufficient cellular material was present in one specimen. In five (39%) of the cases the immunochemical staining supported the light microscopic diagnosis. In four (30%) cases, however, the results indicated that the original light microscopic report was incorrect. Two of these were examples of large cell carcinoma of the lung, in which false negative reports had been issued on pleural fluids. the other two were cases of benign ovarian tumours in which a false positive report had been issued. the immunostaining also clarified the final diagnosis in the three patients (23%) on whom 'suspicious' cytological reports had previously been issued. the remaining case, fluid from a patient with a high grade mixed Mullerian tumour of the ovary, was unresolved. We conclude that immunofluorescent staining by AUA-1, EMA and Ber-EP4 is an aid in the cytological interpretation of serous fluids. CEA is much less helpful.     

Monoclonal antibodies in the cytodiagnosis of serous effusions

In order to assess the value of immunocytochemical staining as a method of discriminating between benign reactive mesothelial cells and malignant epithelial cells in serous effusions, we have studied the reactions of a panel of commercially available antibodies on cells harvested from 83 pleural and peritoneal fluids and compared the results with the clinical and cytological diagnoses. The antibodies used were raised against cytokeratin (PKK1), epithelial membrane antigen (EMA), carcino-embryonic antigen (CEA), pregnancy specific B1-glycoprotein (SP1) and leucocyte common antigen (LCA). Anti-CEA was positive in 16 of 39 effusions (41%) containing carcinoma cells. Pregnancy specific B1-glycoprotein (SP1) was positive in 33% of the same samples. Mesothelial cells did not stain with these antibodies. Thus anti-CEA and SP1 can be used to discriminate between benign mesothelial and malignant epithelial cells in effusions. Anti-PKK1 stained both benign reactive mesothelial cells and malignant epithelial cells and cannot be used to discriminate between these two cell types. Strong positive staining of malignant cells was noted with anti-EMA. However, as occasional weak staining of mesothelial cells was also noted, strong staining with this antibody may be regarded as suspicious but not conclusive of malignancy.     

奶山羊乳腺上皮细胞的分离、培养及鉴定

应用组织块培养法高密度培养、连续传代法建立西农萨能奶山羊乳腺上皮细胞体外培养体系,通过生长曲线绘制、核型分析、免疫荧光染色 (角蛋白、上皮膜抗原、波形蛋白、β-酪蛋白)、油红染色及β-酪蛋白基因的RT-PCR分析进行培养细胞鉴定。实验结果表明细胞生长曲线为典型的S型,染色体数目众数为60,细胞角蛋白、上皮膜抗原、波形蛋白、β-酪蛋白表达均呈阳性,油红染色后可见细胞质内的脂滴,且细胞表达酪蛋白mRNA。说明运用本方法培养的细胞为正常的乳腺上皮细胞,并具有一定的泌乳功能。     

Immunoperoxidase staining of fine needle aspiration specimens previously stained by the Papanicolaou technique

The use of immunoperoxidase techniques was investigated in 21 fine needle aspiration (FNA) cytology smears that had been previously stained by the Papanicolaou technique. The retrospectively selected slides were destained before applying the immunostain, utilizing antisera to calcitonin, prostatic acid phosphatase (PrAP), prostate-specific antigen (PSA), alpha-lactalbumin (AL), S-100 protein (S-100), carcinoembryonic antigen (CEA), common leukocyte antigen (LA), epithelial membrane antigen (EMA) and alpha-fetoprotein (AFP). Positive results were obtained with six of nine small-cell carcinomas of the lung stained with EMA, all three colonic carcinomas stained with CEA, one of two prostatic carcinomas stained with PSA and PrAP, one of two lymphomas stained with LA and the one medullary thyroid carcinoma stained with calcitonin. Negative staining results were observed in the one melanoma stained with S-100, the two breast carcinomas stained with AL and the one hepatocellular carcinoma stained with AFP. These results indicate that immunostaining can be a helpful diagnostic tool in diagnosing some fine needle aspirates using smears previously stained with the Papanicolaou stain.     

Fine needle aspiration as a tool to establish primary human breast cancer cultures in vitro

OBJECTIVE: To verify the potential role of fine needle aspiration (FNA) cytology in obtaining malignant cells from primary breast cancer for establishment of a primary breast cancer cell line. STUDY DESIGN: In four patients with primary breast cancer subjected to FNA for diagnostic purposes, we attempted to establish primary cultures. We successfully obtained one primary cell line, originating in micropapillary invasive breast carcinoma. FNA material obtained under sterile conditions was centrifuged, and the cell pellet was washed with Dulbecco Modified Medium. The resulting suspension was seeded in 25-cm2 tissue culture flasks. The flasks were maintained with released caps in a 37 degrees C incubator with a humidified atmosphere of 5% CO2 in air. After one week, cells attached to the bottom of the flasks and began proliferating. When a culture became confluent, the cells were treated with 0.05% trypsin/0.02% EDTA in a PBS solution and subcultured. The flasks were observed daily with an inverted microscope, and culture passages were performed weekly. RESULTS: The cell line obtained was named I2FPRW and exhibited morphologic and immunohistochemical features of epithelial cells of mammary origin. The cells were positive for cytokeratins (AE1/AE3 and CK 7), EMA and c-erbB-2. At this writing, this cell line was in the 15th passage of subculturing in the flasks with 10% FBS. CONCLUSION: In the present study we demonstrated that is possible to establish a breast cancer cell line from material obtained by FNA cytology. FNA seems to be a valuable method of obtaining malignant cells from breast cancer able to grow free of fibroblasts in cell cultures.     

Identification of Malignant Cells In Serous Effusions Using A Panel of Monoclonal Antibodies Ber-Ep4, Mca-B-12 and Ema

A panel of three monoclonal antibodies (MoAbs) was tested on 29 benign and 53 malignant effusions with the aim of investigating its usefulness for the discrimination between benign and malignant lesions. The panel consisted of MoAbs directed against epithelial membrane antigen (EMA); MCA-b-12, reacting with a 350 kD glycoprotein with mucin-like characteristics present on human breast cancer cells and various other normal and neoplastic tissues, and Ber-EP4, directed against a 34 and 39 kD glycopeptide on human epithelial cells but not on mesothelium. Fifty-two (98%) of the malignant effusions reacted with EMA, 49 (92%) with MCA-b-12 and 44 (83%) with Ber-EP4. Fourteen per cent of benign effusions reacted with EMA, 17% with MCA-b-12 and 7% with Ber-EP4. All seven effusions obtained from patients with a malignant mesothelioma reacted with EMA, six of the seven cases staining intensively. None of the seven stained with Ber-EP4. MCA-b-12 did not react with the cells in one case of malignant mesothelioma. The results suggest that the combination of EMA and Ber-EP4 may be used to discriminate between benign and malignant cells and possibly also between adenocarcinoma and malignant mesothelioma. MCA-b-12 followed in general the reaction pattern of EMA, although often with a less intense staining reaction, making this antibody unsuitable for inclusion in the panel.     

Adenocarcinoma in the rectum of a capped langur (Presbytis pileata)

A rectal adenocarcinoma in a 22-year-old capped langur histologically resembling those in human cases is reported. An ill-defined diffuse tumor with fibrously firm rectal wall showed diffuse infiltrative growth of signet-ring cancer cells. Immunohistochemistry demonstrated positive staining for CEA, lysozyme, EMA, keratin and B72.3.     

Microgravity culture reduces apoptosis and increases the differentiation of a human colorectal carcinoma cell line

Our hypothesis is that rotation increases apoptosis in standard tissue culture medium at shear stresses of greater than approximately 0.3 dyn/cm2. Human MIP-101 poorly differentiated colorectal carcinoma cells were cultured for 6 d in complete medium in monolayers, on Teflon-coated nonadherent surfaces (static three-dimensional [3D]) or in rotating 3D cultures either in microgravity in low-earth orbit (3D microg) or in unit gravity on the ground (3D 1g). Apoptosis (determined morphologically), proliferation (by MIB1 staining), and the expression of epidermal growth-factor receptor (EGF-R), TGF-alpha, or TGF-beta were assessed by immunohistochemistry, while the expression of the differentiation marker carcinoembryonic antigen (CEA) was assessed on Western blots. Over the course of 6 d, static 3D cultures displayed the highest rates of proliferation and lowest apoptosis. This was associated with high EGF-R, TGF-alpha, and TGF-beta expression which was greater than that of a monolayer culture. Both rotated 3D lg and 3D microg cultures displayed lower expression of EGF-R, TGF-alpha, or TGF-beta and proliferation than that of monolayer or static 3D cultures. However, rotated 3D microg displayed significantly less apoptosis and greater CEA expression than rotated 3D 1g cultures. When rotated cultures of MIP-101 cells were grown uncler static conditions for another 3 d, proliferation increased and apoptosis decreased. Thus, rotation appears to increase apoptosis and decrease proliferation, whereas static 3D cultures in either unit or microgravity have less apoptosis, and reduced rotation in microgravity increases CEA expression.     

Fine needle aspiration cytodiagnosis of secretory carcinoma of the breast

Secretory carcinoma (SC) of the breast is a rare variant of breast malignancy and its cytological features in fine needle aspirates have only recently been described. In this communication, our experience with four cases of SC of the breast is presented in which the diagnosis was established on fine needle aspiration cytology (FNAC). In all cases, the samples were cellular and featured diffuse, prominent, intracytoplasmic vacuoles and secretion in malignant cells and occasional signet-ring like forms. The cytodiagnosis of SC in all the cases correlated with subsequent examination of cell blocks of the aspirate and tissue. Cytochemical stains showed diffuse positivity for mucin by alcian blue stain in the vacuolated cells which was periodic acid-Schiff positive and resistant to diastase digestion. Oil-red O staining was negative. Immunopositivity to carcinoembryonic antigen, cytokeratin (CAM 5.2), B72.3 and epithelial membrane antigen was found in malignant cells. The cytodiagnostic criteria for SC of the breast, characteristic cytological features which are useful in a correct FNAC diagnosis and differentiation from other pertinent breast carcinomas, are discussed.