C-Erbb-2 Expression and Dna Ploidy Status In Breast Cancer Cells Obtained By Fine Needle Aspiration (Fna)

The aim of this study was to determine the relative rate of c-erbB-2 oncoprotein immuno-detection on matched fine needle aspiration (FNA) smears and surgical specimens of breast cancer, and to correlate the c-erbB-2 expression with the assessment of the DNA ploidy status. the expression of c-erbB-2 oncoprotein was evaluated using an immunoalkaline phosphatase technique in 49 breast aspirates (four benign and 45 malignant lesions) and 21 matched surgical specimens. the DNA ploidy status was assessed by densitometric techniques on Feulgen-stained smears. Fifty-eight per cent of the smears obtained from 45 malignant lesions and 43% of the 21 corresponding paraffin sections contained cells that were stained by the antibody. the higher incidence of c-erbB-2 expression on smears seems to be due mainly to the better antigen preservation in the fresh cytological preparations. the correlation between c-erbB-2 oncogene expression and DNA ploidy assessment showed an increased incidence of oncogene expression in aneuploid tumours (71% vs 29%; P > 0.05).     

Fine needle aspiration biopsy of hepatocellular carcinoma and hepatocellular nodular lesions: role,controversies and approach to diagnosis

A. Wee
Fine needle aspiration biopsy of hepatocellular carcinoma and hepatocellular nodular lesions: role, controversies and appr oach to diagnosis The role of fine needle aspiration (FNA) biopsy of the liver has evolved. Advances in imaging modalities have obviated the need for tissue confirmation in clinically classic hepatocellular carcinoma (HCC). The risks of needle tract seeding and haematogenous dissemination have been actively debated. Nowadays, cytopathologists are confronted by smaller and smaller nodules, detected due to increased surveillance of high‐risk cirrhotic patients. Tissue characterization of small well‐differentiated hepatocellular nodular lesions (size less than and equal to 2 cm) is extremely challenging and has therapeutic implications. Major issues in the cytodiagnosis of HCC include: (i) distinguishing benign hepatocellular nodular lesions, namely, large regenerative nodules, dysplastic nodules, focal nodular hyperplasia and hepatocellular adenoma from reactive hepatocytes; (ii) distinguishing well‐differentiated HCC from benign hepatocellular nodular lesions; (iii) distinguishing poorly differentiated HCC from intrahepatic cholangiocarcinoma and metastatic carcinomas; (iv) determining the histogenesis of a malignant tumour; and (v) determining the site of origin of a malignant tumour. An overview of the biological evolution and histopathological aspects of dysplastic nodules, small HCCs and ‘nodule‐in‐nodule’ lesions is presented in tandem with clinically relevant nomenclature. An algorithmic approach to FNA diagnosis of HCC and hepatocellular nodular lesions is outlined. Optimal results depend on (i) a dedicated radiologist‐cytopathologist team; (ii) an on‐site cytology service, (iii) a combined cytohistological approach, (iv) immunohistochemistry, and (v) clinicopathological correlation. As we move towards personalized medicine, it is envisaged that hepatic FNA is likely to become a point of care in the management protocol as it takes on the additional role of procurement of tumour and peritumoural tissues for genomic and proteomic profiling to enable targeted molecular therapy.     

Morphometric analysis of AgNORs in thin-layer, liquid-based liver specimens

OBJECTIVE: To detect argyrophilic nucleolar organizer regions (AgNORs) in ThinPrep (Cytyc Corp., Boxborough, Massachusetts, U.S.A.) liver fine needle aspiration (FNA) specimens and to define the diagnostic value of their quantitative analysis in the evaluation of hepatic lesions. STUDY DESIGN: ThinPrep liquid-based FNA biopsy specimens from 49 malignant and benign liver lesions were resampled, fixed in 95% ethanol and stained with the AgNOR technique in accordance with the 1-step colloid method. The specimens included 11 benign and 38 malignant lesions (23 poorly differentiated hepatocellular carcinomas [HCCs] and 15 poorly differentiated metastatic adenocarcinomas [MCs]). Morphometric analysis was performed using a Zeiss Axiolab microscope (Carl Zeiss GmbH, Jena, Germany) with a mechanical stage fitted with a Sony-iris CCD videocamera (Tokyo, Japan). The videocamera was connected to a Pentium III P/C (Intel Corp., Santa Clara, California, U.S.A.) loaded with the appropriate image analysis software. The measurements were performed with ImageScan software (Jandel Scientific, Erkrath, Germany). The number of AgNORs per nucleus (NN) and the total area per nucleus occupied by AgNORs (AR) were calculated semiautomatically. Statistical analysis was performed using the SPSS software package (Chicago, Illinois, U.S.A.). RESULTS: The least significant deviance test for multiple comparisons revealed that NN differed significantly between the 3 groups of samples examined (P < .0001). The mean NN values in HCCs and MCs were significantly different (P < .0001). Logistic regression model demonstrated that as NN increased, the probability of a MC diagnosis decreased (<4%). AR values were different at a statistically significant level only between benign and malignant specimens (P = .00006), not between HCCs and MCs (P = .933). CONCLUSION: Quantitative analysis of AgNORs in ThinPrep specimens could be a diagnostically useful method in liver disease.     

Polyclonal carcinoembryonic antigen staining in the cytologic differential diagnosis of primary and metastatic hepatic malignancies.

In order to assess the utility of immunocytochemical staining of bile canaliculi with a polyclonal antiserum to carcinoembryonic antigen (pCEA) in the differentiation of primary hepatocellular carcinomas from metastatic malignancies, pCEA staining was performed on fine needle aspiration specimens from hepatic lesions in 60 patients. The original cytologic diagnoses were hepatocellular carcinoma in 22 patients, metastatic neoplasm or cholangiocarcinoma in 27 patients and benign hepatocytes in 11 cases. The cytologic diagnoses of malignancy were confirmed by surgical excision, autopsy or clinical investigations in 82% of the patients. Follow-up data, supported by pCEA staining, reversed the original cytologic diagnosis in three cases. Bile canalicular pCEA staining was identified in 18 of 22 cases of hepatocellular carcinoma and in all 11 benign hepatocellular aspirates. All 27 cases of metastatic malignancy or cholangiocarcinoma were negative for canalicular pCEA staining, although 11 cases exhibited cytoplasmic staining. Interpretation of pCEA staining was not affected by the intermingling of malignant cells and benign hepatocytes. Predictive values were 100% for a positive test and 87% for a negative test. These findings indicate that staining with pCEA antiserum is a useful adjunct in the differential cytologic diagnosis of malignant hepatic lesions.     

BCA-225 immunostain: is it of value in liver fine needle aspiration?

OBJECTIVE: To determine the utility of BCA-225 immunostain in differentiating hepatocellular carcinoma (HCC) from other malignant tumors in liverfine needle aspiration specimens. STUDY DESIGN: Cell block materialfrom 87fine needle aspirates of liver lesions, including 25 HCCs, 9 neuroendocrine carcinomas, 12 adenocarcinomas metastatic from the colon, 19 other metastatic adenocarcinomas, 13 other metastatic tumors and 2 cholangiocarcinomas, were immunostained with BCA-225 antibody using both the EnVision avidin-biotin method (Dako U.S.A., Carpinteria, California, U.S.A.), and the traditional avidin-biotin method, without antigen retrieval. Three independent observers evaluated the cases, and a consensus was reached. RESULTS: Strong immunostaining for BCA-225 in at least 10% of malignant cells was considered positive. Two of 25 HCCs, 9 of 19 adenocarcinoma metastases, 1 of 9 neuroendocrine carcinomas, 4 of 13 other metastases and 1 of 2 cholangiocarcinomas were positive. No colon metastases were considered positive. CONCLUSION: BCA-225 may help exclude the diagnoses of metastatic colon adenocarcinoma, neuroendocrine carcinoma and well- to moderately differentiated HCC in this cytologic setting.     

Diagnosis of malignant granular cell tumor by fine needle aspiration cytology:.

BACKGROUND: Malignant granular cell tumors (GCT) are the rarest of all sarcomas, and the histologic differentiation from their benign counterpart may be extremely difficult or impossible unless metastatic disease is demonstrated. To our knowledge, this is the first report of a malignant GCT diagnosed by fine needle aspiration (FNA) cytology. CASE: A 70-year-old, Caucasian female presented with a progressively enlarging left supraclavicular mass. FNA of the mass revealed a metastatic tumor cytologically consistent with GCT. With this diagnosis, a search for other metastatic sites was initiated. Computed tomography (CT) scan revealed several tumor nodules in the lungs and liver. CT-guided FNA and tru-cut needle biopsy of a liver mass confirmed the diagnosis of metastatic GCT. In searching for a primary site, the patient revealed a clinical history of having had a tumor removed from her back two months before; it was reported to be an atypical GCT. Comparison of the three tumors revealed similar histologic, cytologic and immunohistochemical features. CONCLUSION: Evidence of mild to moderate cytologic atypia; increased mitotic activity; locally aggressive growth; increased proliferative activity as demonstrated by immunohistochemical evaluation of proliferation markers; and DNA ploidy analysis, as reported for this case, may be helpful in predicting malignant behavior of GCTs.     

The reliability of microspectrophotometric and flow cytometric nuclear DNA measurements in adenocarcinomas of the breast

The reliability of microspectrophotometric (MSP) and flow cytometric (FCM) nuclear DNA measurements has been studied in 50 human breast adenocarcinomas. The tumor material was obtained by means of fine-needle aspiration biopsy, and all samples except one were found to be highly representative. The results confirm earlier observations that a good correlation exists between modal value (MV) determined by MSP and DNA index (DI) determined by FCM. However, when tumors were classified into low and high malignant variants according to FCM/DI, FCM/S-phase percentages, and MSP histogram types, the concordance was less pronounced. This was found to be due mainly to the fact that in near-diploid tumors a discrepancy exists between MSP and FCM ploidy, as well as between MSP distribution pattern and the estimated percentages of cells in the S-phase region. Another source of discrepancy was observed in tumors with stemlines in the normal tetraploid region, including cells with highly scattered aneuploid DNA values. These tumors were judged by MSP as aneuploid/high malignant and by FCM as euploid/low malignant. In view of this discrepancy, we conclude that the simple determination of the stemline position by MSP/MV or FCM/DI is not sufficient for adequate cytochemical malignancy grading of breast carcinomas. We suggest that a combination of ploidy and percentage of cells scattered outside the modal peaks is a more sensitive method for optimal cytochemical malignancy grading in breast carcinomas.     

Malignant melanoma metastatic to the liver. A cytomorphologic comparative study to identify reproducible diagnostic criteria

OBJECTIVE: To establish cytomorphologic criteria that might facilitate the identification of malignant melanoma (MM) cells with epithelioid (nevoid) morphology, in fine needle aspiration biopsy material from the liver. STUDY DESIGN: Aspirated material from 18 cases of MM with epithelioid features and 24 cases of benign liver lesions (BLL) were examined. The cases were selected based on the availability of corresponding tissue biopsies, adequate cell block material or sufficient number of direct smears to perform immunocytochemical staining. The presence or absence of 7 cytologic criteria were reviewed, and the results were evaluated by multivariate regression analysis. RESULTS: All evaluated criteria were significant for identifying MM cells and differentiating them from reactive hepatocytes (P < .001). Uniform atypia, cell dyscohesion, eccentric nuclei and irregular nuclear membranes supported MM, whereas, monolayered sheets or cordlike arrangement; coarse, granular cytoplasm; and occasional transgressing endothelium in true tissue fragments were evidence of BLL. CONCLUSION: A systematic evaluation of the cytomorphologic features described in this study, in conjunction with the clinical and radiologic findings, can be used to render an immediate, confident and accurate diagnosis of MM metastatic to the liver.     

Silver-binding nucleoli and nucleolar organizer regions in fine needle aspiration cytology of the breast

Silver staining was used to demonstrate nucleoli and nucleolar organizer regions (NORs) in cytopreparations from breast fine needle aspiration (FNA) biopsies, with the resulting black dots being a combination of silver-binding nucleoli and dispersed NORs. Discrete black dots in 200 nuclei were manually counted in 25 silver-stained biopsies; the cytologic diagnoses were made on routine Papanicolaou-stained cytopreparations from the same aspirates. Although malignant breast lesions showed higher counts than did benign breast disease, an overlap of one malignant and three benign lesions occurred. Therefore, the method cannot be recommended for reliably discriminating between malignant and benign routine breast specimens obtained by FNA; it may have some value in assessing tumor behavior.     

Fine needle aspiration cytology of mammary carcinoma with osteoclast-like giant cells

Carcinoma with osteoclast-like giant cells (OCGC) is an uncommon neoplasm characterized by giant cells, prominent vascularization, haemorrhage and areas of cribriform epithelial growth with moderate atypia. Multinucleated giant cells (MGC) have been described in several other breast lesions raising an interesting differential diagnosis, mainly with benign disorders. Due to its rarity few cases have been described cytologically. We retrospectively reviewed 13 fine needle aspiration samples from nine patients with this variant of carcinoma. Nine corresponded to breast tumours and four to axillary, liver, subcutaneous and mediastinal metastatic lesions. The expression of CD68 by giant cells was evaluated immunocytochemically in six cases. All patients had a complete pathological study of the breast neoplasm. Smears showed a double component of epithelial and giant cells. Epithelial clusters were predominantly of intermediate size with irregular contours. Most were cohesive but others showed cellular dissociation with scarce to moderate cellular pleomorphism. Giant cells had well defined, deeply stained cytoplasm and round to elongated morphology. Two metastatic cases were devoid of them. Haemosiderin-laden macrophages were common in smears from breast tumours. In the six cases tested CD68 was expressed in MGC. Cytological features of mammary carcinoma with OCGC correlate closely with the histological ones. Most cases are clearly recognizable as malignant but in others cytological atypia may be minimal, mimicking a benign lesion. In difficult cases the presence of haemosiderin-laden macrophages and the histiocytic nature of the MGC are helpful diagnostic features.     

Fine needle aspiration cytology of bronchioloalveolar-cell carcinoma of the lung

The fine needle aspiration cytology features of twelve peripherally located bronchioloalveolar cell carcinomas of the lung diagnosed by fine needle aspiration biopsy are described. A spectrum of cytomorphologic changes was appreciated, including classic groups showing uniform malignant cells having prominent depth of focus with a lack of significant nuclear molding. Other cells showed features of atypical alveolar macrophages and bronchial-lining cells. The smears demonstrated malignant cells arranged along alveolar septae and possessing hobnail-shaped nuclei. Two cases had associated psammoma bodies, and one case demonstrated optically clear nuclei in the malignant cells. This series stresses the fine needle aspiration features that aid in the recognition of this specific lung neoplasm and its differentiation from benign reactive pulmonary lesions, other primary lung cancers and metastatic tumors.     

Differential diagnosis of malignant intrahepatic tumors by ultrasonically guided fine needle aspiration biopsy and by laparoscopic/intraoperative biopsy. A comparative study

Thirty-six cases of ultrasonographically detected liver tumors (28 malignant and 8 benign lesions) were investigated by both fine needle aspiration (FNA) biopsy and laparoscopic or intraoperative tissue biopsy. The sensitivity of FNA biopsy in detecting a neoplasm was 85.7% while that of tissue biopsy was 82.1%. Specificity was 100% in both methods. A correct histologic diagnosis of the neoplasm could be made on the cytologic smear in only 14 cases (50.0%) due to technical and methodologic difficulties. Endoscopic and intraoperative tissue biopsy yielded sufficient material to make a definite histologic tumor diagnosis in 23 of 28 cases. Additional information on tumor spread and sequelae of liver infiltration was obtained by macroscopic tissue observations in nine cases. These results indicate that FNA biopsy is less sensitive in the classification of primary or secondary liver tumors and that tissue biopsies obtained during laparoscopy or laparotomy are required if a suspected hepatic neoplasm needs to be classified exactly to plan therapy and to establish a prognosis.     

DNA ploidy in colorectal cancer, heterogeneity within and between tumors and relation to survival

Flow cytometry was used to study the incidence of aneuploidy and to determine the significance of multiple sampling from colorectal tumors. DNA ploidy pattern has been proposed as a supplementary prognostic marker, but discrepancies in findings are major. DNA clonal heterogeneity, defined as two or more DNA aneuploid stemlines in the same tumor, is well established. However, most studies have been based on only one biopsy from each tumor. In our study multiple biopsies were taken from 163 patients (88 males and 75 females) electively operated for colorectal cancer. Tumor cells were harvested by fine needle aspiration from fresh frozen biopsies sampled at different sites of each tumor. DNA aneuploidy was detected in tumors from 145 patients (89%), and 18 patients (11%) had a solitary DNA diploid cell population. In a 79 month follow-up period 105 patients had died. Statistical analysis showed that distinction between diploidy and aneuploidy did not predict survival. However, grouping subpopulations into DNA diploid plus near diploid (DNA index (DI) 0. 97-1.15), DNA aneuploid with all aneuploid subpopulations in the interval 1.15-2.06, and DNA aneuploid with subpopulations with DI < 0.97 and/or DI > 2.06, showed a significant difference in survival in a Cox multivariate analysis including Dukes' stage P = 0.049 comparing the second group to the first and P = 0.01 comparing the third group to the first. In 21 (13%) patients only one subpopulation was found, 57 (35%) had two, 44 (27%) had three, and 41 (25%) had four or more different subpopulations. The association of DNA ploidy to survival is shown to be dependent on the number of biopsies analysed.     

Peripheral endothelial cells are not reliable in differentiating primary benign and malignant hepatocellular lesions in fine needle aspirates of the liver

The distinction of hepatocellular carcinoma (HCC) from benign lesions of the liver in fine needle aspiration (FNA) specimens can be problematic. In an attempt to separate well-differentiated HCC from benign hepatocellular lesions, the presence of tissue fragments displaying peripheral endothelial cells (PE) has been proposed in a previous study as a useful feature in favour of malignancy. In this study, we evaluated slides from 59 cases of liver masses undergoing FNA (19 HCC, 40 benign) and evaluated them for the presence of tissue fragments containing PE. We found that 90% of cases of HCC contained tissue fragments in which PE were either focally present or abundant. However, 68% of cases containing only benign hepatocytes also contained tissue fragments in which PE were at least focally present. In addition, it appears that within the group of benign lesions, the presence of PE was related to the overall cellularity of the specimen rather than the specific nature of the lesion. Thus, the presence of PE in tissue fragments does not, in isolation, appear to be a useful morphological feature for the separation of benign and malignant hepatocellular lesions in FNA material.     

Flow cytometric DNA ploidy analysis of soft tissue sarcomas. A comparative study of preoperative fine needle aspirates and postoperative fresh tissues and archival material

Flow cytometric (FCM) DNA ploidy measurements on frozen fresh samples of soft tissue sarcomas were compared with the corresponding analyses on preoperative fine needle aspirates and postoperative formalin-fixed archival tissues from the same tumors. A concordance in ploidy status (diploid versus non-diploid) was obtained for 63% of the fresh tissue-fine needle aspiration (FNA) sample comparisons and for 85% of the fresh tissue-archival material comparisons. The majority of discordances in the fresh tissue-FNA sample comparisons could be explained by FNA sampling errors. In the remaining discordant cases (3 of 27 FNA sample comparisons and 6 of 40 archival material comparisons), sampling errors could not explain the differences in ploidy status. The discordant cases were evenly distributed among the different sampling methods. Method reproducibility was not responsible for the differences in ploidy determinations; tumor heterogeneity may be an explanation for the discrepancies. This study showed that archival soft tissue sarcoma samples are as well suited for DNA ploidy analysis as are fresh frozen tissues.     

Flow cytometric DNA analysis of renal-cell carcinoma. A study of fine needle aspiration biopsies in comparison with multiple surgical samples

The flow cytometric (FCM) DNA analysis of fine needle aspiration (FNA) biopsy material was compared with the DNA analysis of multiple surgical biopsy material from 44 renal-cell carcinomas. Twenty tumors were heterogeneous with respect to their DNA content. Eleven of the 17 tumors that had both diploid and aneuploid tumor cell clones in the surgical specimens gave a diploid DNA content in the aspiration biopsies; the other 6 cases showed aneuploidy in the aspirate. The fine needle aspirates from 18 homogeneously diploid tumors and 9 tumors with an aneuploid DNA content in all eight surgical samples revealed DNA indices similar to those found in the surgical samples. These findings show that aneuploid clones of renal-cell carcinoma can remain undetected by the use of FNA biopsy as a method for obtaining samples for FCM DNA analysis.     

Malignant phyllodes tumor metastatic to the lung with osteogenic differentiation diagnosed on fine needle aspiration biopsy. A case report

BACKGROUND: Osteosarcomatous differentiation in malignant phyllodes tumors is rare. No cases of either primary or metastatic lesions were identified in the literature that were initially diagnosed on fine needle aspiration biopsy. CASE: Cytologic and histologic findings of a metastatic malignant phyllodes tumor with osteosarcomatous differentiation in a 63-year-old woman are presented. This case was diagnosed initially on fine needle aspiration biopsy and confirmed with histologic examination of the pulmonary lesion. CONCLUSION: Although rare, the differential diagnosis of metastatic phyllodes tumor should be considered in the appropriate clinical setting when examining a pleomorphic spindle cell tumor with heterologous elements on fine needle aspiration biopsy.     

The gray zone in breast fine needle aspiration cytology. How to report on it?

OBJECTIVE: To study the "gray zone" in breast fine needle aspiration cytology in which an unequivocal diagnosis cannot be reached with fine needle aspiration cytology findings. STUDY DESIGN: This study compared cytology and histopathology of 72 breast lesions in which an initial cytologic diagnosis of atypia was given. RESULTS: There were 36 benign (50%) and 36 malignant (50%) histologic biopsy cases in the cytologic atypia group. Anisonucleosis, chromatin and nuclear membrane irregularity, and presence of myoepithelial cells were significantly different in benign and malignant cases. CONCLUSION: The gray zone in breast fine needle aspiration cytology is a broad spectrum that changes from proliferative fibrocystic disease to sclerosing adenosis to malignancy. Diagnosing gray zone pathology as atypical in fine needle aspiration cytology causes no delay in treatment as excisional biopsy is recommended for all equivocal cases.     

Fine needle aspiration cytodiagnosis of liver tumors

OBJECTIVE: To present our experience with liver fine needle aspiration (FNA) diagnosis and the adjunctive use of cell blocks with reticulin stain. STUDY DESIGN: The authors reviewed the results of cytopathologic diagnosis obtained by FNA biopsy over a 1-year period, from January 2000 to December 2000, in patients who presented primarily with ultrasonographically suspected liver nodules. FNA smears from 936 patients and cell blocks from 796 patients were reviewed. RESULTS: Among the 936 aspirates studied, the most common malignancy was hepatocellular carcinoma (HCC), which was diagnosed in 427 cases (45.6%), followed by metastatic adenocarcinoma, with 52 cases (5.6%). The concurrent cell block was available in 796 cases. Among them, 574 (72.1%) contained sufficient tissue for diagnosis. Combining analysis of cytologic and histologic specimens, the sensitivity of ultrasound-guided FNA for diagnosis of liver tumors was 85.1% and the specificity 98.7%. The results were better than isolated cytologic analysis, which gave a sensitivity of 78.4% and specificity of 97.4%. The lower diagnostic accuracy of cytology resulted mainly from its lower ability to distinguish well-differentiated HCC from benign lesions. In the cell block sections with reticulin stain, all HCCs showed a decreased or absent reticulin pattern, whereas all the benign hepatocellular lesions usually had a normal trabecular reticulin framework. CONCLUSION: FNA cytology assisted by cell block examination can be an accurate and minimally invasive method for the definitive pathologic diagnosis of primary benign and malignant liver masses and for confirmation of tumors metastatic to the liver. In addition, reticulin staining should be part of the routine assessment of cell blocks. It enhances diagnostic accuracy, particularly for well-differentiated HCC.     

Acral myxoinflammatory fibroblastic sarcoma in FNAB samples: can we distinguish it from other myxoid lesions?

We analysed cytomorphological characteristics of three fine needle aspiration biopsy (FNAB) samples of acral myxoinflammatory fibroblastic sarcoma (AMIFS) as well as the features of a number of other benign and malignant myxoid lesions. The analysis showed that FNAB samples from two cases of AMIFS had similar cytomorphology, containing all the characteristic features described in surgical biopsies: myxoid material, spindle cells with bipolar cytoplasmic extensions, epithelioid cells with globules of extra-cellular material, ganglion-like and lipoblast-like giant cells. Only the inflammatory component was scarce. The third sample did not contain features typical of AMIFS. Samples from other myxoid tumours resembled AMIFS to some extent, however, none of them contained all three tumour components characteristic of AMIFS. Cytomorphology of AMIFS may be characteristic enough to enable a definitive diagnosis from FNAB, provided all the distinctive features are sampled.