Cytophotometrical and biochemical characterization of nonpalpable, mammographically detected mammary adenocarcinomas

Fifty nonpalpable, mammographically detected, invasive breast carcinomas were analysed with respect to DNA distribution pattern, steroid receptor content, and histopathological criteria. No significant histomorphological differences were found as compared to palpable breast carcinomas. In contrast, DNA distributions of palpable and nonpalpable tumors differed. Ninety percent of these relatively small breast carcinomas were found to exhibit nuclear DNA amounts within the diploid and tetraploid regions of normal breast epithelium. In earlier findings in palpable breast carcinomas, 55% are of the diploid-tetraploid type. The mean cellular content of the estrogen receptor was 1.0 fmole/microgram DNA in this group of mammographically detected carcinomas, which is significantly higher than in routinely detected, ie, larger, breast carcinomas. It is suggested that, despite the histomorphological findings, nonpalpable, mammographically detected breast carcinomas are dominated by biologically highly differentiated, slowly proliferating carcinomas with a favourable prognosis.     

Fine needle aspiration cytology in the work-up of mammographic and ultrasonographic findings in breast cancer screening: an attempt at differentiating in situ and invasive carcinoma.

This study evaluated the results of fine needle aspiration cytology (FNAC) from the first four years of organized mammography screening for breast cancer in Oslo, particularly our policy in differentiating in situ and invasive carcinoma. Lesions were aspirated directly, ultrasound guided, by stereotaxic device or biopsy localization plate. All lesions were aspirated by cytopathologists working with the radiologists at the breast diagnostic centre. Smears were evaluated immediately for assessment of adequacy and a preliminary diagnosis was given to the surgeon. When FNAC revealed malignancy, diagnostic terms were as follows: (1) invasive carcinoma; (2) ductal carcinoma in situ of comedo type (high nuclear grade), cannot evaluate infiltration; (3) ductal carcinoma in situ of low nuclear grade and (4) papillary tumour, cannot evaluate infiltration. There were 953 cases, 70% of which were nonpalpable. Insufficient material was obtained in 5.8%. Absolute and complete sensitivity were 81% and 91%, respectively. Specificity was 85%. There were 448 histologically proven carcinomas. 383 of these were invasive. 362 carcinomas (in situ and invasive) (80.8%) were diagnosed directly on FNAC. Distinction between invasive and in situ carcinoma was possible in 294 of 320 directly diagnosed invasive carcinomas (91.8%). PPV of a diagnosis of invasive carcinoma was 97%. Our data showed that definitive cytological diagnosis of invasive carcinoma was possible in more than 90% of fully diagnostic smears and allowed definitive primary surgery in these women.     

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.     

Cytologic grading and DNA image cytometry of breast carcinoma on fine needle aspiration cytology smears

OBJECTIVE: To correlate the cytologic grade of breast carcinoma with DNA image cytometry (ICM) and nuclear area on fine needle aspiration cytology (FNAC) smears. STUDY DESIGN: In this prospective study, FNAC material from 28 breast carcinomas were studied for cytologic grade and DNA ICM. Breast carcinomas were classified as grade 1-3 (low to high). DNA histograms were classified by the modified Auer method. Degree of hyperploidy (DH), ploidy balance (PB) and nuclear area (NA) were measured on Feulgen-stained smears by a CAS 200 image cytometer. Cytologic grade was correlated with DNA ICM findings and NA. RESULTS: There were 3 cytologic grade 1, 13 grade 2 and 12 grade 3 breast carcinomas. Seven of eight cases of hypertetraploid aneuploidy were grade 3 tumors. All cytologic grade 1 tumors were diploid. There were significant differences in DH, PB and NA in different grades of breast carcinoma (one-way ANOVA). CONCLUSION: DNA image cytometry in combination with cytologic grading might offer additional information for the characterization of breast carcinomas diagnosed by FNAC. These observations are of particular interest with the introduction of preoperative chemotherapy.     

Stereotaxic fine needle aspiration cytology of clinically occult malignant and premalignant breast lesions

Stereotaxic fine needle aspiration (FNA) cytology was used to study clinically occult (nonpalpable) breast lesions in 114 consecutive patients with mammographically suspicious findings prior to excisional biopsy. The aspirate contained insufficient material for cytologic evaluation in 15 cases (13.2%), which were histologically diagnosed as benign (7 cases), atypical hyperplasia (7 cases) or carcinoma in situ (1 case). The cytologic findings indicated a benign lesion in 77 cases (67.5%), which were histologically diagnosed as benign (71 cases) or atypical ductal hyperplasia (6 cases). The cytologic sample showed atypia in eight cases (7.0%), which were histologically diagnosed as severe atypical ductal hyperplasia (three cases), carcinoma in situ (one case) or proliferative fibrocystic disease (four cases). In the eight cases (7.0%) cytologically interpreted as probably malignant, histology confirmed six invasive carcinomas, one carcinoma in situ and one fibrocystic disease. Of six cases (4.4%) cytologically reported as malignant, five were histologically diagnosed as invasive carcinoma and one as carcinoma in situ. Overall, stereotaxic FNA cytology reported as malignant or probably malignant 14 of the 15 cases with a histologic confirmation of malignancy, for a sensitivity of 93.3%. Cytology correctly identified 78 of the 83 histologically negative cases, for a specificity of 94.0%. The 16 cases histologically diagnosed as ductal hyperplasia, which carries a high risk for subsequent malignancy, were studied in detail in an effort to define histologic and cytologic criteria for this entity. Using selected histologic criteria, 11 of these cases were graded as showing mild-to-moderate atypical hyperplasia and 5 as showing severe atypical hyperplasia. Three of the latter cases were similarly identified by an analogous cytologic grading; the other two cases had insufficient cytologic samples. The total results in this series of 114 cases support the use of stereotaxic FNA cytology in the diagnosis of these nonpalpable breast lesions, examples of which are illustrated. In particular, it may help to raise the low specificity yielded by mammography alone, which would represent a significant advance for the patient in terms of the accuracy, expediency and reduced cost of diagnosing these lesions.     

Fine needle aspiration cytology of thymic tumors

Cytologic material was reviewed from 23 mediastinal tumors clinically suspected as thymomas. The thymomas had a characteristic biphasic cell pattern in material obtained by fine needle aspiration biopsy that was easy to recognize and possible to differentiate from carcinoid tumors, malignant lymphomas and oat-cell carcinomas.     

Value of aspiration cytology of the thyroid in metastatic disease

Metastases may simulate primary malignant tumors of the thyroid, causing problems in the diagnosis and management of patients with a history of cancer. In the seven-year period of July 1978 through June 1985, 8 of 549 needle aspirates of the thyroid contained metastatic tumor, 6 of which were subsequently confirmed by histologic study. The primary sites of origin were the breast, kidney, colon and stomach as well as lymphoma. The cytologic features observed in the aspiration biopsy material from the six cases were characteristic of each of the primary tumors. Three of the patients had had prior resections of carcinomas (breast, colon and stomach) while in three patients the cytologic diagnosis of the thyroid aspirates led to the discovery of the primary tumor (kidney and two lymphomas). One case of lymphoma/leukemia and one case of previously biopsied lung carcinoma were confirmed on clinical grounds. It is of critical importance that primary thyroid neoplasms occurring in patients known to have primary tumors elsewhere be distinguished from disseminated tumors involving the thyroid. Our experience suggests that fine needle aspiration is of considerable value in this differential diagnosis. Needle aspirates of the thyroid are also of value in leading to the diagnosis of unsuspected nonthyroidal primary cancer.     

High-throughput molecular analysis from leftover of fine needle aspiration cytology of mammographically detected breast cancer

We investigated whether residual material from diagnostic smears of fine needle aspirations (FNAs) of mammographically detected breast lesions can be successfully used to extract RNA for reliable gene expression analysis. Twenty-eight patients underwent FNA of breast lesions under ultrasonographic guidance. After smearing slides for cytology, residual cells were rinsed with TRIzol to recover RNA. RNA yield ranged from 0.78 to 88.40 μg per sample. FNA leftovers from 23 nonpalpable breast cancers were selected for gene expression profiling using oligonucleotide microarrays. Clusters generated by global expression profiles partitioned samples in well-distinguished subgroups that overlapped with clusters obtained using "biologic scores" (cytohistologic variables) and differed from clusters based on "technical scores" (RNA/complementary RNA/microarray quality). Microarray profiling used to measure the grade of differentiation and estrogen receptor and ERBB2/HER2 status reflected the results obtained by histology and immunohistochemistry. Given that proliferative status in the FNA material is not always assessable, we designed and performed on FNA leftover a multiprobe genomic signature for proliferation genes that strongly correlated with the Ki67 index examined on histologic material. These findings show that cells residual to cytologic smears of FNA are suitable for obtaining high-quality RNA for high-throughput analysis even when taken from small nonpalpable breast lesions.     

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.     

Fine needle aspiration biopsy of neuroendocrine breast carcinoma metastatic to the thyroid. A case report

BACKGROUND: Tumors showing neuroendocrine differentiation arise in a wide range of organs, and metastatic neuroendocrine tumors may be difficult to differentiate from primary tumors. This report describes an unusual case of metastatic breast carcinoma with neuroendocrine differentiation that presented as a solitary thyroid nodule. The diagnosis was made by fine needle aspiration biopsy (FNAB). CASE: A 52-year-old woman presented with a thyroid nodule and bilateral enlarged supraclavicular fossa lymph nodes. FNAB revealed a neuroendocrine carcinoma. Further questioning revealed that the patient had had a breast carcinoma resected eight years previously. The diagnosis of metastatic neuroendocrine breast carcinoma was established by immunocytochemistry. The patient received antiestrogen therapy but subsequently developed skeletal metastases. CONCLUSION: Neuroendocrine carcinomas from various sites show similar cytologic features. In this case, a diagnosis of breast carcinoma metastatic to the thyroid was suggested by the clinical history and confirmed by FNAB with immunocytochemistry.     

DNA ploidy assessment method applied to primary breast carcinoma FNA biopsies

The goal of our study was to assess suitability of FNA biopsy material as a source of samples (cell suspension) for DNA ploidy assessment in neoplastic tumors using flow cytometry. DNA ploidy is an established prognostic factor in many types of cancers. Aneuploid breast tumors are characterized by increased aggressiveness which manifests itself through rapid local progression and metastatic spread. Investigated specimens were breast cancer FNA biopsy cell suspensions. Measurements were performed using flow cytometry. Material studied comprised 143 cases analyzed in 1999-2000. We found in this group 101 carcinoma cases with aneuploid type and 42 cases of primary breast carcinoma with diploid type of cell cycle. Immunocytochemical assesssment of estrogen receptor and progesterone receptor status was performed in group of 105 cases. DNA ploidy was compared to receptor status of the investigated cells. DNA aneuploidy correlated with weak or no reaction for the presence of estrogen and progesterone receptors. Our study demonstrates the suitability of DNA ploidy assessment method applied to cytological material from FNA biopsies.     

Fine needle aspiration biopsy of the breast. Influence of the number of passes and the sample size on the diagnostic yield

The sensitivity of fine needle aspiration (FNA) biopsy of the breast as a function of the number of aspirations performed on any given lesion was investigated. Four separate aspirations each were performed on over 400 lesions of the breast, 93 of which yielded a cytologic diagnosis. The incremental diagnostic yields of each subsequent aspiration were tabulated. The first aspirate of the sequence gave the greatest yield, with smaller incremental yields on the second through the fourth aspirates. Benign and malignant lesions gave similar results, as did palpable and nonpalpable lesions (the latter being usually smaller in size). A mathematical extrapolation of the data indicates that three or four aspirations of any given lesion provide the optimal yield within the limits of practicality. This performance of multiple FNA biopsies is particularly important when the pathologist does not perform the biopsy, or is unable to assist in the immediate interpretation of the specimen to assess its adequacy.     

Clinical significance of pepsinogen C tumor expression in patients with stage D2 prostate carcinoma

Pepsinogen C is an aspartyl protease mainly involved in the digestion of proteins in the stomach, and an androgen-inducible protein in breast cancer cells. The aims of this study were to evaluate the expression and clinical significance of this enzyme in the primary tumors of prostate cancer patients with bone metastasis who were scheduled to receive antiandrogenic therapy. This study was prospectively performed in 28 stage D2 prostate cancer patients who, after diagnosis, received maximum androgen blockade. Pepsinogen C tumor expression was analyzed in samples (24 from needle biopsy cylinders and four from transurethral resection specimens) from primary tumors using an immunohistochemical assay. Twelve prostate carcinomas (42.8%) were positive for pepsinogen C. Pepsinogen C was a significant prognostic factor to predict a longer overall survival in the patients of our study (p<0.01). Pepsinogen C can be a new prognostic factor and a useful biological marker of androgen dependency in prostate cancer.     

Dilution effect of nontumor cells on flow cytometric DNA S-phase fraction in breast cancer fine needle aspirates

OBJECTIVE: To analyze the proportion of nontumor cells in fine needle aspirates of breast carcinoma and its influence on flow cytometric S-phase fraction (SPF) estimation. STUDY DESIGN: We analyzed the proportion of nontumor cells in fine needle aspiration biopsy smears, performed flow cytometric analysis of DNA ploidy and SPF on freshly aspirated tumor material and analyzed histograms manually and automatically using Multi-Cycle AV software (Phoenix Flow Systems, San Diego, California, U.S.A.) for cell cycle analysis. We corrected SPF of diploid tumors for the dilution effect using an individually established percentage of nontumor cells (individual correction) and the mean proportion of nontumor cells in diploid tumors (factor correction). RESULTS: The proportion of nontumor cells ranged from 0.5% to 76.6% (mean, 12.6; SD, 15.7) in 55 diploid tumors and from 0.5% to 53% (mean, 8.6; SD, 8.9) in 84 aneuploid tumors (p=0.178). In 14 of 139 (10%) samples, the proportion of nontumor cells exceeded 20%. The mean SPF values of diploid tumors without correction were 4.9% (manually) and 6.5% (automatically) and of aneuploid tumors, 9.5% and 11.0%, respectively. In univariate Cox survival analysis, noncorrected SPF was a significant prognostic factor in overall survival (p < 0.001). Neither individual nor factor correction of SPF significantly changed its prognostic value. CONCLUSION: Fine needle aspirates contain low proportions of nontumor cells, having an insignificant dilution effect on SPF estimation. Most probably, SPF could be reliably estimated usingfreshly aspirated tumor material without any correction or adjustment.     

Accuracy of touch imprint cytology of image-directed breast core needle biopsies

OBJECTIVE: To evaluate the accuracy of touch imprint (TI) cytology of image-directed core needle biopsy (CNB) specimens of nonpalpable breast lesions. STUDY DESIGN: Fifty-two consecutive CNBs from 44 patients were performed under mammographic or ultrasound guidance. Air- dried TIs of CNBs were stained with Diff-Quik. TI cellularity was considered adequate if six or more ductal cell groups were identified. CNBs and TIs were interpreted in a blinded fashion. RESULTS: TI cellularity was adequate in 37/52 (71%) cases, including 17/20 carcinomas and 20/32 benign lesions. Among 17 carcinomas, TIs were positive in 12, suspicious in 4 and atypical in 1. One case of lactational change was suspicious on TI, and 5/8 fibroadenomas were atypical. No benign lesions were called "carcinoma" on cytology. When lesions categorized as "carcinoma" or "suspicious" were considered positive and those classified as "atypical" or "benign" were scored as negative, TI sensitivity and specificity were 94% and 95%, respectively. When the "atypical" cases were grouped with the positive cases, TI sensitivity was 100%, with 75% specificity. CONCLUSION: With satisfactory cellularity, TIs of CNBs are highly accurate. When immediate evaluation of CNB specimens is important, TIs can potentially decrease the number of biopsy passes required and provide preliminary diagnoses.     

Diagnostic value of flow cytometric DNA determination combined with fine needle aspiration biopsy in thyroid tumors

The nuclear DNA content and the numbers of cells in the S and G2M phases of the cell cycle were determined by flow cytometry (FCM) in fine needle aspirates of 187 thyroid tumors to evaluate the diagnostic value of nuclear DNA content determination in combination with aspiration cytology. DNA aneuploidy was present in 4 of 5 follicular carcinomas, 2 of 3 anaplastic carcinomas, 5 of 15 excised follicular adenomas and 2 of 20 excised adenomatous goiters; all 7 papillary carcinomas and 4 lymphomas were diploid in the aspirate. Aneuploid carcinomas had easily distinguishable S and/or G2M phases, unlike the benign aneuploid tumors. None of the histologically benign tumors or the nonexcised tumors had greater than 6% S-phase cells, and only one benign tumor had greater than 9% G2M-phase cells. In contrast, all lymphomas had greater than 10% S-phase cells and four of seven papillary carcinomas had greater than 9% G2M-phase cells. The use of FCM determination in combination with fine needle aspiration biopsy cytology improved the diagnostic potential of the latter technique.     

Determination of nuclear DNA content and hormone receptors in breast cancer by the CAS 100 cell analysis system as related to morphologic grade and biochemical results

Quantitative DNA analysis by the CAS 100 Cell Analysis System was performed on 120 cases of primary breast carcinoma using touch preparations from fresh biopsy specimens in 110 cases and archival, restrained fine needle preparations in 10 cases. Fifteen cases of metastatic breast carcinoma and 15 cases of benign breast lesions were also analyzed. Overall, 76.7% of the carcinomas examined were aneuploid, with most DNA indices between 1.6 and 2.0. DNA anomalies were strongly related to nuclear atypia but not to structural differentiation. The hormone receptor content, when compared with DNA data and morphologic features, emerged as a biologically independent factor. Agreement between quantitative immunocytochemical assay (QICA) using the CAS system and traditional dextran-coated charcoal assay (DCCA) in discriminating positive and negative status for estrogen receptors and progesterone receptors was 86% and 82%, respectively. Marked variations, however, occurred in the numerical values. Considering the advantages of QICA and the importance of tumor heterogeneity in particular, the use of traditional DCCA as the reference technique and only guide for therapy no longer seems justified.     

Fine needle aspiration of palpable breast lesions. Results obtained with cytocentrifuge preparation of aspirates

The use of cytocentrifugation in the preparation of fine needle aspiration (FNA) specimens from the breast was evaluated. A total of 174 fine needle aspirates of breast masses were flushed into cytospin Collection Fluid, from which Papanicolaou-stained Cytospin preparations were made in the laboratory. Comparison of these preparations to conventional smears of aspirates showed no significant differences in the number or morphology of the cells. In 148 cases, the FNA cytologic study was followed up by excisional biopsy, Tru-Cut biopsy and/or a combination of mammography and clinical follow-up of two to nine months. Of 36 verified carcinomas, 31 (86%) were correctly diagnosed, with a zero false-positive rate. Among the 74 cytologically benign aspirates, 2 carcinomas were found on open biopsy, giving a false-negative rate of 3%. Lipomas were not diagnosable with this technique. This technique should be considered in institutions with a high turnover of junior staff members, which frequently results in a higher number of poorly smeared specimens or in poorly fixed/air-dried specimens that give suboptimal results with the Papanicolaou stain. With this method, there is less risk of creation of potentially hazardous aerosols and further preparations for additional studies may be made if required.     

DNA polymerase chain reaction using fine needle aspiration biopsy smears to evaluate non-Hodgkin's lymphoma.

OBJECTIVE: To apply polymerase chain reaction (PCR) analysis to the fine needle aspiration biopsy (FNAB) evaluation of lymphoid proliferations. STUDY DESIGN: We analyzed 37 consecutive archived FNAB malignant lymphoma specimens. Immunophenotypic data from the fine needle aspiration biopsy and excisional biopsy material was available for all specimens. PCR to identify monoclonal rearrangements of the immunoglobulin heavy chain gene, T-cell receptor and translocations involving the bcl-1 and bcl-2 genes was performed. RESULTS: Seventy-eight percent of cases were detected by at least one of these assays. Where DNA analysis was performed on excisional biopsy material, 70% of the cases had identical results; no discordant results for the immunoglobulin heavy chain gene or T-cell receptor were found. In 23% of cases, after review of all available data, a discordant result was thought to be a consequence of a false negative result in DNA analysis of excisional biopsy material. CONCLUSION: These findings indicate that PCR analysis of archived FNAB material, when necessary, provides useful information for diagnosis and staging of malignant non-Hodgkin's lymphomas.     

Metastatic tumors in fine needle aspiration biopsy of the thyroid

A series of almost 25,000 thyroids examined by fine needle aspiration (FNA) biopsy was reviewed to ascertain the incidence and presentation of metastatic cancers in thyroid FNA samples. Metastatic cancers in FNA samples from the thyroid were identified in 25 cases (0.1%); the primary tumors were carcinomas of the kidney (8 cases), lung (7 cases), breast (5 cases), cervix uteri (1 case) and colon (1 case) and 1 case each of malignant melanoma, malignant pleural mesothelioma and rhabdomyosarcoma. FNA cytology was positive in all 25 cases. In 11 cases, the primary tumor was clinically known at the time of FNA biopsy; of the other 14 cases, cytology suggested that the malignancy was metastatic in only 5. Metastases of renal and mammary adenocarcinomas were almost indistinguishable from follicular and papillary thyroid carcinomas on cytologic grounds. The results demonstrate the rarity of this finding and the difficulty of diagnosing a metastatic tumor in the thyroid by FNA biopsy, in the absence of a clinical history of a prior primary neoplasm.