Identification of putative oncogenes in lung adenocarcinoma by a comprehensive functional genomic approach.

Amplification and overexpression of putative oncogenes confer growth advantages for tumor development. We used a functional genomic approach that integrated simultaneous genomic and transcript microarray, proteomics, and tissue microarray analyses to directly identify putative oncogenes in lung adenocarcinoma. We first identified 183 genes with increases in both genomic copy number and transcript in six lung adenocarcinoma cell lines. Next, we used two-dimensional polyacrylamide gel electrophoresis and mass spectrometry to identify 42 proteins that were overexpressed in the cancer cells relative to normal cells. Comparing the 183 genes with the 42 proteins, we identified four genes - PRDX1, EEF1A2, CALR, and KCIP-1 - in which elevated protein expression correlated with both increased DNA copy number and increased transcript levels (all r > 0.84, two-sided P < 0.05). These findings were validated by Southern, Northern, and Western blotting. Specific inhibition of EEF1A2 and KCIP-1 expression with siRNA in the four cell lines tested suppressed proliferation and induced apoptosis. Parallel fluorescence in situ hybridization and immunohistochemical analyses of EEF1A2 and KCIP-1 in tissue microarrays from patients with lung adenocarcinoma showed that gene amplification was associated with high protein expression for both genes and that protein overexpression was related to tumor grade, disease stage, Ki-67 expression, and a shorter survival of patients. The amplification of EEF1A2 and KCIP-1 and the presence of overexpressed protein in tumor samples strongly suggest that these genes could be oncogenes and hence potential targets for diagnosis and therapy in lung adenocarcinoma.

Detection of chromosome 11q13 breakpoints by interphase fluorescence in situ hybridization. A useful ancillary method for the diagnosis of mantle cell lymphoma.

We assessed cytologic specimens from 11 mantle cell lymphomas (MCLs) and 32 other B-cell non-Hodgkin lymphomas (NHLs) for 11q13 breakpoints using a 2-color fluorescence in situ hybridization (FISH) assay that uses an 11q13 probe centered on the CCND1 gene and a centromeric chromosome 11 probe (CEP11). The number of nuclei in 200 cells were counted, and results were expressed as an 11q13/CEP11 ratio. All MCLs showed a high percentage of interphase nuclei with 3 or more 11q13 signals (mean, 74.8%; range 57%-90%). In contrast, in other B-cell NHLs the mean percentage of cells with 3 or more 11q13 signals was 9.2%. All MCLs had an elevated 11q13/CEP11 ratio (mean, 1.38). The mean ratio for other B-cell NHLs was 0.99. Two non-MCL cases, 1 large B-cell and 1 B-cell unclassified NHL, had high 11q13/CEP11 ratios of 1.15 and 1.30, respectively. Conventional cytogenetic analysis performed on the former case revealed a t(5;11)(q31;q13). Interphase FISH analysis using 11q13 and CEP11 probes is a convenient ancillary method for assisting in the diagnosis of MCL. This commercially available assay is simple to use on cytology or imprint specimens, and results can be obtained within 24 hours.

Indeterminate fine-needle aspiration biopsy of the thyroid: identification of subgroups at high risk for invasive carcinoma.

BACKGROUND: We examined the various cytologic features of indeterminate thyroid fine-needle aspirates along with known clinical and radiologic risk factors to determine whether any parameters were predictive of malignancy. METHODS: Indeterminate fine-needle aspirates were prospectively categorized into four subgroups: (1) suspicious for papillary carcinoma, (2) follicular neoplasm, (3) Hürthle cell neoplasm, and (4) hypercellular follicular aspirates with colloid. Several clinical risk factors were examined, and subgroup comparisons were performed with Fisher's exact test. RESULTS: Of 571 fine-needle aspirate cytologic findings 104 were interpreted as indeterminate for malignancy, and 81 patients underwent thyroidectomy. Invasive cancer was diagnosed in 9 of 10 lesions cytologically suspicious for papillary carcinoma, 8 of 43 follicular neoplasms, 5 of 18 Hürthle cell neoplasms, and 0 of 10 hypercellular aspirates. Cytologic subgroup (p < 0.0001) and age of 50 years or older (p = 0.008) were the only significant predictors of malignancy. When used together, age of 50 years or older and a cytologic diagnosis of follicular or Hürthle cell neoplasm also identified a subgroup of patients at high risk (9 of 20) of invasive malignancy (p = 0.01). CONCLUSIONS: The majority of invasive cancers (18 of 22, 82%) were found in patients whose lesions were suspicious for papillary carcinoma or in patients 50 years or older with follicular or Hürthle cell neoplasms. The risk of carcinoma in these combined subgroups (18 of 30, 60%) warrants early surgical intervention.

Poorly-differentiated adenoid cystic carcinoma: cytologic appearance in fine-needle aspirates of distant metastases.

Adenoid cystic carcinoma (ACC) is a primary salivary-gland neoplasm which typically yields characteristic cytomorphology upon fine-needle aspiration (FNA). We report on the FNA findings of a case of ACC metastatic to the liver which demonstrated a predominantly solid, poorly-differentiated pattern, an unusual but well-recognized subtype associated with a poor clinical outcome. The FNA findings in 7 additional cases of ACC metastatic to distant sites were also reviewed, with 4 cases displaying a prominent poorly-differentiated component. These findings suggest that, although not commonly recognized in salivary-gland FNAs, the poorly-differentiated pattern of ACC does occur in metastatic deposits and should be recognized as such, thereby preventing a needless search for a second primary malignancy.

Use of fine-needle aspiration biopsy in the evaluation of splenic lesions in a cancer center.

Fine-needle aspiration biopsy (FNAB) of the spleen was performed on 50 patients, of whom 40 had had a previous diagnosis of malignancy (23 lymphoproliferative disorders, 13 carcinomas, 3 melanomas, and 1 sarcoma). The cytologic diagnoses included 22 cases positive for malignancy (10 lymphomas, 9 metastatic carcinomas, 2 metastatic melanomas, and 1 sarcoma), 18 cases negative for malignancy, 4 cases suspicious for malignancy, and 6 nondiagnostic specimens. No major complications were associated with the FNAB procedure, however, one patient did develop a pneumothorax that resolved spontaneously. Subsequent splenectomy was performed in 10 of the 50 cases. There were no false-positive diagnoses, and only one false-negative diagnosis, which was attributed to sampling error. The aspirate, showing only benign splenic parenchyma, was from a patient with splenomegaly and no previous diagnosis; subsequent splenectomy showed acute myelogenous leukemia. In our study, FNAB proved to be a safe and valuable diagnostic tool for evaluating splenic lesions in oncologic patients.

Fine-needle aspiration biopsy of metastatic small cell carcinoma from extrapulmonary sites.

Like a pulmonary counterpart, extrapulmonary small cell carcinoma (SCC) is an aggressive tumor with a high rate of metastasis. Forty-nine fine-needle aspiration biopsies (FNABs) (36 patients) of various primary sites other than the lung diagnosed as metastatic SCC (including Merkel cell carcinoma) were reviewed. FNABs were derived from lymph nodes (20), liver (7), bone (2), breast (1), pancreas (1), and skin/soft tissue (18). Primary tumor sites included the prostate (14), skin (11; Merkel cell carcinoma), cervix (5), urinary bladder (3), urethra (1), ovary (1), and parotid (1). Aspirates revealed predominantly dispersed single tumor cells with occasional clustering. Tumor cells were small with scant cytoplasm, fine powdery chromatin, and inconspicuous nucleoli. Nuclear molding, mitotic figures, and apoptotic bodies were frequently observed. In four cases, findings from the FNABs were used to render the initial diagnosis of SCC. FNAB is useful for determining whether metastases contain a SCC component, a finding that may alter clinical management. Cytologically, SCC from different primary sites cannot be differentiated, and its distinction requires clinical and radiographic correlation.

Diagnostic pitfalls in thyroid fine-needle aspiration: a review of 394 cases.

To determine the diagnostic pitfalls of thyroid fine-needle aspiration (FNA), we reviewed 394 thyroid aspirates obtained between January 1986 and December 1990. Surgical follow-up was available for 150 aspirations. The cytologic diagnoses were categorized into four groups: benign, 57; indeterminate, 51; malignant, 33; and nondiagnostic specimen, nine. There were three false negative diagnoses (3%), which upon review were judged to be inadequate specimens. Three false positive diagnoses (7%) were identified: in the first two cases, follicular adenomas were mistaken for papillary carcinoma; in the third case, atypical Hürthle cells were mistaken for a Hürthle cell carcinoma. Our results showed a sensitivity of 93% and a specificity of 91% for the detection of malignancy. If indeterminate cases were considered positive, the specificity decreased to 50%, while the sensitivity increased to 97%. We conclude that: 1) certain follicular adenomas may display cytologic features mimicking papillary carcinoma; 2) as in follicular neoplasms, aspirates of Hürthle cell adenomas cannot be differentiated from Hürthle cell carcinomas; 3) with adequate sampling, false negative results can be markedly reduced.

Fine-needle aspiration cytology of seminoma: a review of 16 cases.

We report the cytomorphologic features of 16 fine-needle aspiration (FNA) biopsies of seminoma obtained from 16 male patients. The aspirates included two primary gonadal tumors (one occurring in a cryptorchid testis), two primary mediastinal tumors, and 12 metastases (two with unknown primaries). Analysis of the aspirates revealed a primarily dispersed cell population of large cells with scant to moderately abundant cytoplasm. The nuclei were round to slightly irregular, had finely granular chromatin, and had either one central prominent nucleolus or two to three smaller nucleoli. Variable numbers of lymphocytes and plasma cells were intermingled with the tumor cells. Only a few cases had epithelioid histiocytes or the characteristic "tigroid" background. The cytologic features of the metastases were distinctive and were considered diagnostic for therapeutic management. In six cases, an initial diagnosis of seminoma by FNA biopsy identified the neoplasm as germ cell in origin rather than other neoplasms in the differential diagnosis, thereby expediting therapeutic management.

Blastic variant of mantle-cell lymphoma: cytomorphologic, immunocytochemical, and molecular genetic features of tissue obtained by fine-needle aspiration biopsy.

Mantel-cell lymphoma (MCL) is a rare type of non-Hodgkin's lymphoma that has a moderately aggressive clinical course, generally between that a low-grade and intermediate-grade lymphomas. However, a small subset of MCLs, the so-called "blastic" variant, exhibits a poor prognosis and an aggressive clinical course. We describe a case of blastic MCL that occurred in a 64-yr-old man and that was diagnosed and accurately subclassified as blastic MCL on the basis of an fine-needle aspiration (FNA) biopsy. The aspirate smears showed a monotonous population of intermediate-sized lymphocytes with irregular nuclear contours, finely dispersed nuclear chromatin, and inconspicuous nucleoli. Material was obtained by FNA for ancillary studies (immunocytochemical stains, flow cytometry, cytogenetics, image analysis, and molecular studies) that supported the diagnosis of blastic MCL. Surgical biopsy confirmed the diagnosis. These findings underscore the utility of FNA in diagnosing lymphomas, particularly when the cytomorphologic examination is combined with appropriate ancillary studies.

Diagnosing intramuscular myxoma by fine-needle aspiration: a multidisciplinary approach.

The gross and microscopic appearances of aspirates from ten intramuscular myxomas are reported. The specimens were obtained from seven women and three men, ages 43 to 75, who had tumors involving the muscles of the thigh (7), upper arm (2), and forearm (1). Magnetic resonance (MR) imaging performed in six of the ten cases revealed well-defined, sharply demarcated tumors exhibiting low signal intensity relative to muscle on the T1-weighted images. The tumors were hyperintense to muscle on T2-weighted images. All aspirated tissues were clear, tenacious, and viscous. Smears contained few spindled and histiocytoid cells in an abundant mucoid background. Spindle cells demonstrated long cytoplasmic processes that in areas intertwined to form fibrillar tangles. Nuclei were oval to spindled with fine chromatin and inconspicuous nucleoli. Capillaries were sparse with simple (non-plexiform) branching. The differential diagnosis of myxoid lesions of the extremities includes benign entities such as myxoid schwannoma and neurofibroma, mesenchymal repair, and ganglion cyst, as well as malignant neoplasms such as myxoid liposarcoma, fibrosarcoma, malignant fibrous histiocytoma, and extraskeletal chondrosarcoma. The findings of this study revealed that, although the cytologic features were suggestive of intramuscular myxoma, a definitive diagnosis was often difficult, owing to scant cellularity and lack of distinctive cytologic features. The MR imaging findings may be utilized as an adjunct to the cytologic features to more confidently suggest a diagnosis of intramuscular myxoma.

Cytologic findings of collecting duct carcinoma of the kidney.

We report the cytologic features of eight fine-needle aspirations (FNA) and eight exfoliative specimens of collecting duct carcinoma (CDC) obtained from six patients. The four men and two women ranged in age from 27 to 69 years (mean = 45 yr) and all had advanced stage disease at presentation (one stage III, five stage IV). Five of the six patients died of widespread disease, and one is alive and well (mean survival, 28 mo; range, 11-48 mo). The smears of the FNA and exfoliative specimens were scantly to moderately cellular. Tumor cells showed moderate pleomorphism and were arranged primarily in cohesive groups that rarely had a papillary configuration. Nuclei had irregular nuclear contours, coarse chromatin, and one to three nucleoli. In the majority of cases the cytoplasm was finely vacuolated, and occasionally there were large intracytoplasmic vacuoles. Intracytoplasmic mucin was demonstrated in two aspirates. Psammoma bodies were present in four of the seven fluids. In two patients, the cytologic diagnosis was supported by positive immunostaining for high-molecular-weight keratin and Ulex europaeus agglutinin I lectin. Leu M-1 was focally positive in one case and negative in the other. The cytologic features of CDC were readily identified as malignant; however, they were not distinctive and overlapped with those of high-grade renal cell carcinoma with papillary features and transitional cell carcinoma.

Fine-needle aspiration cytology of recurrent and metastatic mixed mesodermal tumors.

We report the cytological and clinical findings of 16 fine-needle aspirates (FNAs) performed on recurrent (n = 6) and metastatic (n = 10) mixed mesodermal tumors (MMMTs). The median interval between the primary diagnosis and FNA was 16 mo. Primary sites were the endometrium (n = 11), the ovary (n = 3), the cervix (n = 1), and pelvic soft tissue (n = 1). Primary tumors showed carcinoma with homologous mesenchymal components in 13 cases and focal heterologous elements in three (two chondrosarcomas and one rhabdomyosarcoma). The FNAs showed carcinoma in all 16 cases, with adenocarcinoma differentiation in three. Mesenchymal elements were identified in aspirates of three recurrent and two metastatic lesions. They were all homologous. aspirates. We conclude that mesenchymal components in FNAs of MMMTs are less likely to be seen in metastatic lesions, and that heterologous mesenchymal components are rarely seen in these aspirates even in recurrent disease. These findings confirm that the epithelial component is responsible for the malignant behaviour of MMMTs, and suggest that these lesions may need to be classified as sarcomatoid carcinomas rather than true carcinosarcomas.

Metastatic small-cell carcinoma of the prostate diagnosed by fine-needle aspiration biopsy.

Fourteen fine-needle aspiration biopsies (FNABs) of metastatic small-cell carcinoma done on 12 patients who had histologically documented primary small-cell carcinoma of the prostate are described. The FNABs were of lymph node (four cases), liver (four cases), bone (two cases), pancreas (one case), perirectal soft tissue (one case), perineum (one case), and lung (one case). One patient underwent three FNABs. No patient had a second primary tumor elsewhere. Cytologic smears were cellular with numerous single tumor cells, many apoptotic bodies, and variable numbers of mitotic figures. Tight cell clusters with molded nuclei and finely stippled chromatin were seen in all cases. An organoid pattern of tumor cells was seen focally in two cases. Features distinguishing small-cell carcinoma from poorly differentiated prostate carcinoma were cell size, finely stippled chromatin, inconspicuous nucleoli, and numerous single tumor cells. Distinction from small-cell carcinoma of other primary sites requires clinical and radiologic correlation. We conclude that cytologic specimens are useful for documenting metastatic small-cell carcinoma of the prostate and for differentiating between it and conventional prostate carcinoma in metastatic sites.

Percutaneous needle biopsy diagnosis of benign neurogenic neoplasms.

Preoperative diagnosis of benign neurogenic neoplasms (BNNs) provides useful information in guiding management. To assess the effectiveness of fine-needle aspiration (FNA) and needle core biopsy (NCB) in diagnosing schwannomas and neurofibromas, 40 percutaneous biopsies interpreted as BNNs or obtained from lesions subsequently shown by excision to be BNNs were reviewed. The 13 aspirates diagnostic of BNN revealed spindle cells arranged haphazardly in irregular tissue fragments and in parallel as elongated ropy fascicles, with a myxoid to fibrillary background. The nuclei were buckled, often with intranuclear cytoplasmic inclusions. Four lesions showed nuclear pleomorphism without mitoses. Of 19 schwannomas evaluated by FNA, four (21%) were diagnosed as schwannomas and seven (37%) as BNNs. Ten neurofibromas were aspirated, revealing two (20%) BNNs. Of seven nondiagnostic FNAs accompanied by NCB, three (43%) indicated a BNN. The sensitivities of FNA, NCB, and both modalities in diagnosing BNNs were 43,60, and 71%, respectively. For the 16 FNAs showing features of BNNs, subsequent excisions revealed 11 schwannomas, two neurofibromas, one neurogenic sarcoma, one fibromyxoid neoplasm of uncertain malignant potential, and one unclassified low-grade myxoid sarcoma. FNA can be effective in diagnosing BNNs. If collagenous or myxoid lesions yield paucicellular nondiagnostic aspirates, NCB is helpful. Lowgrade sarcoma and neurofibromatous areas of neurogenic sarcoma may be misinterpreted as BNNs by percutaneous biopsy. BNNs may show nuclear pleomorphism without mitotic activity, and should not be mistaken for sarcoma.

Concomitant lymphoma and metastatic carcinoma in a lymph node: diagnosis by fine-needle aspiration biopsy in two cases.

Concomitant lymphoma and metastatic carcinoma are an unusual occurrence in a lymph node. We report two patients in whom synchronous malignancies were diagnosed by fine-needle aspiration biopsy (FNAB). In one case, the FNAB diagnoses of both small lymphocytic lymphoma and metastatic breast carcinoma were the initial diagnoses. In the second case, metastatic poorly differentiated squamous carcinoma was an unexpected finding in a patient with a history of small lymphocytic lymphoma. The aspirates in both cases showed two distinct cell populations, one consisting of a dispersed population of small uniform lymphoid cells and the other comprising large atypical single cells and cohesive clusters of epithelial cells. In both cases, the cytologic diagnoses were supported by immunohistochemical and flow cytometric studies.

Fine-needle aspiration of intra-abdominal desmoplastic small cell tumor.

The cytologic features of four cases of histologically confirmed intra-abdominal desmoplastic small cell tumor (DSCT) that occurred in males between 18-27 yr of age are presented. Smears showed small cells with scant cytoplasm which were primarily arranged in loosely cohesive clusters. Nuclei were oval to round with evenly distributed, finely granular chromatin and inconspicuous nucleoli. As is typical of DSCT, the cells strongly expressed keratin and desmin in all cases. In the one case studied by electron microscopy, it was demonstrated that the cells were joined by small junctions and contained paranuclear aggregates of intermediate filaments. The absence of the characteristic desmoplastic stroma in DSCT aspirates and the nonspecific cytologic features of this small round-cell tumor (SRCT) made cytologic interpretation difficult. Cytopathologists should be aware of this entity so that DSCT is included in the differential diagnosis of SRCTs that occur in intra-abdominal sites. A panel of markers that includes keratin and desmin should be used to assist in the identification of DSCT.

Cytology of malignant melanoma of soft parts: fine-needle aspirates and exfoliative specimens.

We describe the morphologic features of 25 cytology specimens (13 fine-needle aspirates and 12 exfoliative specimens) obtained from nine patients with malignant melanoma of soft parts (MMSP). Analysis of the fine-needle aspirates and exfoliative specimens revealed primarily a dispersed cell population with occasional cell clustering. Tumor cells were round to polygonal with moderately abundant cytoplasm and had round nuclei with prominent nucleoli. In two cases, an initial definitive diagnosis of MMSP was rendered on material obtained by fine-needle aspiration with the aid of immunocytochemical and ultrastructural studies.

Quality-assurance study of simultaneously sampled, non-correlating cervical cytology and biopsies.

As part of a quality assurance study, we reviewed 223 cases of simultaneously sampled cervical smears and biopsies that showed a significant lack of correlation for squamous dysplasia or carcinoma. In 153 of the 223 cases (68.6%), the cytology was negative and the biopsy positive. After review of the specimens, errors in this group were found to be of the following types: sampling 64%, interpretive 29%, and combined sampling and interpretive 7%. In the remaining 70 cases (31.4%), the biopsy was negative and the Papanicolaou smear positive. In these cases, the following types of errors occurred: sampling 54%, interpretive 33%, and combination 13%. Twenty-nine of these 70 patients showed dysplasia on follow-up material. These findings indicate there are a significant number of false-negative Papanicolaou smears, mostly because of sampling problems. There are few false-positives. In cases of positive Papanicolaou and negative biopsy, dysplasia is likely to be present in subsequent samples.

Myoepithelioma of the parotid. Report of a case initially examined by fine needle aspiration biopsy.

Fine needle aspiration (FNA) biopsy of a large, deep lobe of a parotid mass revealed a uniform population of small, spindled cells with no distinctive cytologic features. Biopsy and later resection of the mass revealed a myoepithelioma. Myoepithelioma is a rare lesion, occurring most commonly in the parotid gland. To date, the FNA description of myoepithelioma has been limited to one report of a lesion in the breast. We report the FNA findings of this tumor arising in the parotid, with histologic and electron microscopic confirmation.

Fine needle aspiration cytology of extramedullary chronic myelogenous leukemia.

Chronic myelogenous leukemia (CML) shows extramedullary involvement in 10% of cases. We report the cytologic findings of fine needle aspiration (FNA) of recurrent CML in extramedullary sites in 11 patients with CML. The patients' ages ranged from 24 to 62 years (median, 38 years). There were seven male and four female patients. The aspiration sites were mostly lymph nodes (cervical in 7, retroperitoneal in 2, axillary in 1) and abdominal wall soft tissue (1). The numbers of blasts in the aspirates ranged from 27% to over 90%. Confirmation of the myeloid nature of the blasts was done using naphthol AS-D chloroacetate esterase in one case. Cytologic and flow cytometric immunotyping was done in eight cases. Two cases were based on cytomorphologic features only. Two of the eight immunophenotyped aspirates showed evidence of T-lymphoblastic differentiation. Another showed a mixed myeloid and T-cell phenotype. Blasts were seen in the peripheral blood and bone marrow in 4 of the 11 patients. We thus conclude that extramedullary involvement by CML in our series was associated with younger age, high incidence of cervical lymphadenopathy, increased blasts and frequent lack of bone marrow and peripheral blood involvement. T-cell phenotypes appeared to be higher in our series than reported in the literature. This suggests that there is a need for phenotyping some aspirates of recurrent extramedullary CML, mainly to evaluate the possibility of dedifferentiation and its possible impact on the behavior of the neoplasm.