Implementation of DNA cytometric measurements in fine-needle aspiration biopsy diagnostics of breast disease

BACKGROUND: For this study, the diagnostic protocol included fine-needle aspiration biopsy (FNAB) and DNA cytometric measurements. The objective of the study was to improve the diagnostic sensitivity of FNAB. METHODS: Sixty-eight FNAB samples were stained using the Feulgen stain, and DNA histograms were produced by selecting the nuclei of cell groups and free cells separately. RESULTS: Among 28 samples that were classified as definitely benign (n = 17 samples) or atypical but benign (n = 11 samples), DNA cytometry generally showed diploid/peridiploid peaks. Three of those samples were associated later with carcinoma. In those samples DNA cytometry did not improve sensitivity. Among moderately atypical samples (n = 17 samples), 8 samples were diagnosed later as carcinoma. DNA cytometry helped the diagnostic procedure in 62.5% of those samples. Among 21 samples that were classified as highly suspicious (n = 10 samples) or definitely malignant (n = 11 samples), DNA cytometry generally showed atypical DNA histograms (20 of 21 histograms), and DNA cytometry supported the diagnosis of carcinoma in 95.2%. Histograms that were based on free cells frequently were more abnormal compared with histograms that were based on cell groups. Histologically verified benign lesions also could show abnormalities in DNA histograms. Accepting wider gates than are used normally for primarily Feulgen stained samples on these restained samples resulted in improved specificity, efficiency, and predictive values. CONCLUSIONS: DNA cytometry has a potential to support the differential diagnosis of breast lesions, and sampling of free cells increases sensitivity. Benign breast lesions (fibrocystic disease, fibroadenoma) included DNA-cytometrically abnormal cell clones and showed tendencies toward polyploidy, which should be included in the diagnostic criteria.     

Laser scanning cytometry for the detection of neoplasia in urologic cytology specimens

BACKGROUND: The objective of the current pilot project was to assess the efficacy of laser scanning cytometry (LSC) for DNA ploidy analysis of atypical urologic cytology specimens to enhance the distinction between benign and malignant changes. METHODS: Forty selected urologic cytology specimens that previously had been categorized as normal, atypical, or malignant were studied. Nuclear propidium iodide and fluorescence intensity measurements were converted to pixel values, which were used to create scattergrams that excluded debris and cell clusters from ploidy analysis, creating a gated (isolated) region of predominantly single cells for LSC ploidy analysis. Integral histograms then were created to show the number of cells present in diploid, tetraploid, and aneuploid peaks; these histograms also were used to assess DNA ploidy. RESULTS: Ten normal specimens, 10 malignant specimens, and 20 atypical specimens were examined to assess the efficacy of LSC ploidy analysis. Normal and malignant specimens generated reference histograms for comparison with the atypical specimens and exhibited 90% specificity and 100% sensitivity. Ten atypical aneuploid specimens had histogram and scattergram patterns similar to those produced by malignant specimens and, using the cytometer's relocation feature, the presence of atypical cells was confirmed in the aneuploid regions. CONCLUSIONS: The authors determined that DNA ploidy analysis of atypical urologic cytology specimens using LSC is a useful adjunct tool for identifying malignant specimens that lack sufficient cytologic criteria for diagnosis by light microscopy alone. However, LSC is time consuming and requires expensive equipment.     

Prognostic significance of DNA image cytometry in libyan breast cancer

Background: We evaluated the relation of nuclear DNA content and clinicopathological features and prognosis in primary breast cancer of female Libyan patients with variable stage and grade and different treatment regimes. Patients and Methods: Histological samples from 104 patients of breast carcinoma were retrospectively studied by computerized nuclear DNA cytometry. Isolated nuclei from paraffin sections were stained with Feulgen stain and DNA was measured using a computer-assisted image analysis cytometry system. In each case, 200 nuclei were measured and the DNA histograms, S phase fraction (SPF) and number of cells above 5c and 9c were determined. We applied different approaches in the analysis of DNA to compare the DNA histograms with different clinicopathological features and survival. Results: The mean of DNA ploidy mode for all tumors was 3.43; 82.7% of tumors were aneuploid and 17.3% were diploid. The median SPF was 3.5% for DNA diploid and 13.5% for DNA aneuploid tumors. DNA aneuploid tumors and high SPF were associated with advanced stage, distant metastasis, high histological grade and lymph node involvement. The SPF was also associated with large tumor size and with younger patients (<50 years). In the overall population (median follow-up 51 months), patients with aneuploid DNA histograms and high SPF values had shorter survival times than those with diploid DNA histograms and low SPF values (p = 0.001, p < 0.0001, respectively). Also, short survival was associated with a multiploid DNA histogram and with DNA aneuploid cells ≥5 cells (p < 0.0001, p = 0.001, respectively). In a Cox multivariate analysis, DNA ploidy (p = 0.010), age (p = 0.038) and clinical stage (p = 0.001) were independent predictors of overall survival, and DNA ploidy (p = 0.018) and clinical stage (p = 0.001) also proved to be independent predictors of disease-specific survival. The SPF cutoff point of 11% might be applied to separate patients into good and poor prognosis groups. Conclusions: DNA image cytometry with careful analysis of the histograms may provide valuable prognostic information in Libyan breast cancer, with potential clinical implications in patient management, particularly in predicting the patients at high risk for metastasis and recurrence who should be considered as candidates for combined adjuvant therapy.     

Flow cytometric analysis of breast needle aspirates

This study investigated two hypotheses: (1) sufficient cells may be obtained by needle aspiration of breast nodules to produce good flow cytometric DNA profiles; and (2) benign breast lesions do not produce aneuploid G0G1 peaks, and therefore a distinct aneuploid peak is sufficient for a diagnosis of malignancy. Breast specimens received in Surgical Pathology between December 1985 and February 1987 were aspirated, and the cells stained with propidium iodide for flow cytometric DNA analysis. A total of 344 specimens were aspirated, of which 204 (59%) were malignant and 140 (41%) benign. One hundred fifty-three malignant and 111 benign specimens contained sufficient cells for analysis. Cytologic smears were available for 177 malignant and 123 benign specimens. DNA histograms were considered diagnostic of malignancy if an aneuploid peak was present which contained at least 20% of the cells in the distribution, and had a DNA index greater than or equal to 1.2. Using these criteria, 73 of 153 (48%) carcinomas could be identified. None of the benign lesions satisfied these criteria. One fibroadenoma with atypical hyperplasia produced a distinct peak which contained less than 5% of the cells in the histogram, and had a DNA index of 1.25. Flow cytometric analysis provides objective data that complement the subjective cytologic interpretation of fine needle aspirates.     

Flow cytometry and Feulgen cytophotometry in evaluation of effusions

Fifty-eight effusions (42 pleural and 16 ascitic fluids) from patients with and without cancer were analyzed by conventional cytology and the results compared with DNA patterns generated by flow cytometry of 10(4) nuclei and several modes of Feulgen cytophotometry. In 31 patients (24 without evidence of cancer and seven with history of cancer and cytologically negative fluids), the fluids were diploid by flow cytometry. One fluid with atypical cells from a lymphoma suspect was also diploid. Flow cytometry of 26 cytologically cancerous fluids disclosed aneuploid DNA patterns in 16 and diploid patterns in ten. Feulgen cytophotometry of 11 of these fluids (three aneuploid, eight diploid) was performed on nuclear preparations identical to those used in flow cytometry and on restrained smears used for visual analysis. The analysis was performed in two modes: as a study of 500 sequential nuclei in an automated system, mimicking flow cytometry, and visually selected large, presumably malignant nuclei. In nine of the 11 cases, the DNA content of visually selected cancer cells was aneuploid, even though this DNA pattern was not evident in the analysis of 500 sequential cells. In two cases, both diploid by flow cytometry, the Feulgen analysis confirmed the presence of cancer cells in the diploid range. In samples of 10(4) nuclei representing a mixed population of cells occurring in effusions, the presence of aneuploid cancer cells may not be disclosed by conventional flow cytometry. A larger sample of cells, a detailed analysis of DNA histograms, and perhaps sorting of select cells in the hypertetraploid range, may prove essential before flow cytometry can be accepted as a diagnostic tool in the laboratory in the assessment of effusions.     

Evaluation of DNA ploidy and degree of DNA abnormality in benign and malignant melanocytic lesions of the skin using video imaging

BACKGROUND: Making a morphologic distinction between benign and malignant melanocytic tumors of the skin is frequently difficult, especially because "gray zones" between these lesions often exist. DNA image cytometry as an adjuvant method for the diagnosis and prognostic prediction of premalignant lesions and malignant tumors of many other organs is already well established. The aim of this study was to determine whether DNA image cytometry is helpful in distinguishing benign from malignant melanocytic lesions and whether cytometry would give valid information with which to predict the prognoses associated with malignant melanomas. METHODS: DNA image cytometry was performed on 127 benign and 58 primary maligant melanomas of the skin as well as 11 metastatic melanomas, using an enzymatic single cell solution according to a method described by Heiden et al. in Cytometry (1991;12:614-21). RESULTS: DNA aneuploidy was graded by DNA index (DI) and a 2c deviation index (2cDI). In contrast to benign melanocytic lesions (with 16% DNA aneuploidy), primary and metastatic malignant melanomas had significantly higher frequencies of DNA aneuploidy (86% and 73%, respectively). In the degree of DNA aneuploidy, significant differences between benign and malignant melanocytic tumors could be observed. The mean 2cDI of aneuploid benign lesions was 1.0, whereas the primary malignant melanomas had a mean 2cDI of 2.92 and the metastatic melanomas a mean of 6.9. The frequency of DNA aneuploidy increased with Breslow thickness. Twenty-one patients with primary malignant melanoma developed metastases. All metastasizing primary tumors were aneuploid and showed a significantly higher grade of DNA aneuploidy than nonmetastasizing malignant melanomas. Moreover, none of the diploid malignant melanomas developed metastases. CONCLUSIONS: This study reveals that DNA image cytometry is prognostically and diagnostically relevant to the evaluation of melanocytic lesions of the skin. Nevertheless, it cannot be relied on alone to provide enough information for a diagnosis.     

胸水脱落细胞DNA含量及VEGF,p53和CEA表达在良恶性胸腔积液鉴别中的意义

目的：探讨检测胸水细胞的DNA倍体以及VEGF、p53、CEA表达鉴别良恶性胸腔积液的价值。方法：73例胸腔积液患者，良性组13例，恶性组60例。图像细胞光度技术（image cytometry，ICM）进行DNA含量检测，免疫组化Envision法检测VEGF、p53、CEA。结果：良性胸水DNA异倍体率仅占23．1％。恶性胸水的异倍体率占77．8％，两者有统计学差异（P=0．001）。良性胸腔积液患者VEGF、p53、CEA表达率分别为12．5％、0、0。恶性胸腔积液VEGF、p53、CEA表达率分别为17．5％，17．5％和68．4％，与良恶性胸腔积液中CEA的表达相比有统计学差异（P〈0．001），VEGF和p53的表达无显著差异（P=0．722，P=0．198）。经DNA倍体联合VEGF、p53、CEA检测，敏感率可达90．4％，特异度87．5％，符合率90％。结论：ICM检测胸水细胞的DNA倍体联合VEGF、p53、CEA表达值得进一步探讨．有可能成为鉴别良恶性胸腔积液的又一方法。     

Early diagnosis of cutaneous T-cell lymphoma by DNA flow cytometry on skin biopsies

Seven patients with mycosis fungoides early plaque stage with nondiagnostic histology had single-cell DNA content measured by flow cytometry for estimation of clonal ploidy. A total of 63 skin specimens were examined by histology and DNA measurements concurrently during the course of the disease. In addition, six patients had blood samples studied. All seven patients demonstrated aneuploid DNA histograms when the specimens were obtained from skin lesions. In 36 specimens the aneuploid peaks were hyperdiploid. By sequential studies one patient demonstrated two different aneuploid cell clones, one located in the hyperdiploid region and one located in the hypotetraploid region. All patients developed mycosis fungoides which were histologically confirmed, and the time from first aneuploid DNA histogram until diagnostic histology varied from 5 to 21 months (median, 12 months). In six of the patients a normal diploid DNA histogram was found of peripheral blood lymphocytes. The finding of aneuploidy in patients with early mycosis fungoides who still have a nondiagnostic histology emphasizes the value of flow cytometry as a complementary diagnostic aid which facilitates an early diagnosis.     

DNA image analysis combined with routine cytology improves diagnostic sensitivity of common bile duct brushing

BACKGROUND: Cytologic evaluation of common bile duct brushings has a low sensitivity for diagnosing malignancy because of scant cellularity, poor cellular preservation, or sampling errors occur. The aim of this study was to evaluate whether cytology combined with image analysis improves the diagnostic accuracy of bile duct brushing in comparison with cytology alone. METHODS: Forty-nine specimens of bile duct brushings obtained from 45 patients during endoscopic retrograde cholangiopancreatography were evaluated using cytology and image analysis. Specimens were classified as negative, atypical, suspicious, or malignant by using cytologic evaluation. DNA histograms were classified as diploid (D), broad diploid (BD), aneuploid (A), or tetraploid (T). Degree of hyperploidy (DH), representing cells with a DNA content > 5C was evaluated using a cutoff value of > or = 1%. Final diagnosis of cancer was based on tissue specimens that were obtained by fine-needle aspiration or surgical biopsy and clinical fol- low-up. RESULTS: Thirty-four patients ultimately proved to have a malignancy. Cytology revealed 19 negative cases, 15 atypical cases, 9 suspicious cases, and 6 malignant cases. Together, suspicious and malignant cytology cases yielded a sensitivity of 44% and a specificity of 100% for a cytologic diagnosis of cancer. The DNA histogram pattern was D in 24 cases, BD in 9 cases, and A in 16 cases. BD and A patterns were significantly associated with malignancy (P < 0.001). A DH > or = 1% was noted in 22 cases. DH alone had a sensitivity of 62% and a specificity of 91% and was significantly associated with malignancy (P < 0.004). Atypical cytology alone had a false-negative rate of 29%, but in combination with a DH > or = 1%, the false-negative rate decreased to 7%. Additionally, when the authors combined atypical, suspicious, and malignant cytology with a DH > or = 1%, the diagnostic sensitivity increased to 88%, but the specificity decreased to 73%. CONCLUSIONS: Combined cytology and image analysis of bile duct brushing increased diagnostic sensitivity compared with cytology alone. The findings suggest that image analysis may help select patients having atypical cytology who should undergo a more rigorous evaluation for malignancy. A larger prospective study of the usefulness of combined cytology and image analysis of bile duct brushing is warranted.     

Diagnostic approach of effusion cytology using computerized image analysis

The objective of this study is to investigate whether image cytometry is a sensitive and specific method for the differential diagnosis of equivocal cells in routine cytology of effusion smears. One hundred four effusion smears were studied from routine cytologic material. Cytologically 56 (53.8%) of the smears were classified as malignant, 26 (24%) as suspicious and 22 (21.1%) as benign. Two morphometric variables (nuclear major axis length and nuclear area) of the nuclei were measured by an image analysis system. Higher values for the area were found for malignant rather than benign and suspicious cells (p < 0.0005 and p < 0.005 respectively). The same result was extracted for the nuclear major axis length values (p < 0.0005 and p < 0.0005 respectively). Values of nuclear major axis length and nuclear area didn't differ significantly between benign and suspicious cells (p = 0.071 and p = 0.066 respectively). The results show that the range of the values for suspicious cells is closer to the range of the benign cells. Cytomorphometry of the effusion smear cells may provide important information for the differentiation of atypical mesothelial cells from malignant adenocarcinoma cells.     

Flow cytometrically determined DNA content of breast carcinoma and benign lesions: correlations with histopathological parameters

The relative DNA content of cellular samples from 54 patients affected by breast carcinomas and 20 affected by benign breast lesions (including 11 fibroadenomas) was measured by flow cytometry. All normal tissue samples and 17/20 (85%) specimens from benign lesions exhibited a cytometrically diploid DNA distribution, 3/20 (15%) benign lesions an abnormal DNA content, and 35/54 (65%) carcinomas at least one aneuploid cell subpopulation. Furthermore, 9/54 (17%) tumors were characterized by the presence of more than one aneuploid cell subpopulation. The results also indicate that flow cytometry can be used to recognize lymph nodes infiltrated by aneuploid cells. Statistically significant correlations were evidenced between the occurrence of aneuploidy or the ploidy level measured as DNA index and the nodal infiltration status. The percentage of S cells can also be extracted from DNA content distribution histograms. Statistically significant differences (p less than 0.01) were also observed for the percentage of S cells between normal tissues (6.2 +/- 3.2 SD) and benign lesions (11.1 +/- 6.6 SD), normal tissues (6.2 +/- 3.2 SD) and aneuploid tumors (19.7 +/- 10.3 SD), benign lesions (11.1 +/- 6.6 SD) and aneuploid tumors (19.7 +/- 10.3 SD), and diploid (7.9 +/- 4.0 SD) and aneuploid tumors (19.7 +/- 10.3 SD).     

Study of DNA content using a flow cytometry method and steroid hormone receptors in breast cancer

DNA level measured by flow cytometry and estrogen and progesterone receptors assayed in tissue samples obtained from 85 malignant and 16 benign lesions of the breast. All the benign tumors revealed 2c DNA content and most of them were receptor-negative, while 74.1% of breast carcinomas displayed aneuploidy. Three patients (3.5%) had two lines of aneuploid cells. Many aneuploid tumors were receptor-negative. Preoperative radiation treatment (14-20 Gy) did not significantly influence the level of steroid hormone receptors in tumors. Estrogen receptor level was higher in menopausal patients than in premenopausal ones.     

Oncocytic tumours of the salivary gland, kidney, and thyroid: nuclear DNA patterns studied by flow cytometry

Nuclear DNA ploidy studies were performed by flow cytometry on extracted nuclei from 12 oncocytic tumours of the salivary gland, 65 oncocytic tumours of the kidney, and 37 oncocytic tumours of the thyroid gland from the pathology archives of the Mayo Clinic. In order to provide an interesting clinical spectrum, three different classes of well-differentiated oncocytic tumours were selected for examination. Salivary gland oncocytic tumours were chosen for their generally benign behaviour. Oncocytic thyroid cancers exhibiting malignant potential because of local invasion, were thought to represent the opposite extreme of aggressiveness. Renal oncocytic tumours were known to demonstrate an intermediate degree of malignancy. All of the oncocytic salivary gland tumours showed a 'normal' DNA histogram and had a benign clinical course. For the oncocytic tumours of the kidney, 45% of DNA histograms were normal, 40% exhibited a significant increase in the DNA tetraploid/polyploid (4C) peak, and 15% showed a DNA aneuploid peak. Three patients with a DNA tetraploid pattern developed tumour metastasis and two have died from metastatic renal cancer. Among the oncocytic thyroid cancers, 27% were normal, 22% exhibited an increased DNA tetraploid peak, and 51% had a distinct DNA aneuploid peak. None of the thyroid tumour patients with a normal DNA pattern or with an increased DNA tetraploid peak died as a result of thyroid malignancy. In contrast, 58% of patients whose thyroid tumours showed a DNA aneuploid peak subsequently died from thyroid cancer.     

Aneuploid cells in benign breast-lesions monitored by flow-cytometry

It has been observed that various types of benign breast disease are associated to an increased risk of breast cancer. The biological significance of this association remains unclear: both benign and malignant lesions could independently have a common set of risk factors. The cellular DNA content of biopsy samples from 47 breast benign lesions was analyzed by flow cytometry. Flow cytometric measurements evidenced that 11/47 cases showed at least one aneuploid cell subpopulation. The presence of aneuploid subpopulations in benign lesions could be related to an unknown cellular alteration predisponding the developement of benign and malignant lesions independently.     

Oncocytic tumors of salivary gland type: A study with emphasis on nuclear DNA ploidy

We studied nine cases of oncocytic tumors of salivary gland type in order to evaluate their clinico-pathologic profiles and nuclear DNA patterns as criteria for the differential diagnosis between benign and malignant forms. The tumors were located at the parotid gland, palate, and orbit. The age of the patients ranged between 35 and 85 years and the sex ratio (F:M) was 1:0.8. Seven tumors were capsulated and typical cytology: they were composed of polyhedrical cells with large, eosinophilic, and granular cytoplasm and dark nucleus. The remaining two tumors exhibited malignancy criteria for the oncocytic tumors: atypia, pleomorphism, and mitosis. The evaluation of the nuclear DNA content was also distinct: benign tumors had a DNA diploid pattern and the malignant neoplasms displayed a DNA aneuploid pattern. These observations point to DNA nuclear assessment as an additional criterion to discriminate neoplasms with divergent clinical behavior and prognosis. © 1993 Wiley-Liss, Inc.     

DNA image cytometry is a useful adjunct tool in the prediction of disease outcome in patients with stage II and stage III colorectal cancer

BACKGROUND: We assessed the prognostic value of the nuclear DNA content measured in the primary tumours of 123 patients with stage II or stage III colorectal cancer (CRC). METHODS: Isolated nuclei from paraffin sections were stained with the Feulgen reaction, and DNA was measured using a computer-assisted image analysis cytometry system. We applied 4 different approaches in the analysis of DNA histograms: the ABCDE approach, histogram range, peak evaluation and DNA cut-off values. RESULTS: Using the histogram range, the narrow range was rare (3.7%) in patients who died of disease (n = 28) as compared with 16.4% among those alive (n = 74; p = 0.017). Modal peak evaluation was a significant predictor of disease-free survival (DFS; Kaplan-Meier log-rank p = 0.0235). In the range evaluation, the 1st set (low-start gates) was a significant predictor of DFS (log-rank p = 0.0121), where disease recurrence was closely associated with the widest range (1.8->10c; c = haploid DNA content) gates. Recurrence-free survival was 3 times better in narrow-gate histograms than wide-range histograms (p < 0.03). The 1st set also proved to be a significant predictor of disease-specific survival (DSS; log-rank p = 0.0045), which was markedly better (77.8-90.0%) among the patients with the narrow-gate histograms. Grading of the histogram range into 2 categories (with 6.0c as cut-off), was a powerful predictor of both DSS (log-rank p = 0.0092) and 5-year DFS (p = 0.0106) in the whole series, and separately in stage III (but not stage II) disease, with p = 0.0131 and p = 0.0201, respectively. CONCLUSION: The DNA image cytometry with careful analysis of the histograms may provide valuable prognostic information in CRC, with potential clinical implications in patient management, particularly in predicting the patients at high risk for recurrence who should be considered as candidates for adjuvant therapy.     

Image and flow DNA cytometry of small cell carcinoma of the lung

Both image and flow DNA cytometry were performed in isolated nuclei from paraffin-embedded tumor tissue of patients with small cell carcinoma of the lung (SCCL). In 14 patients tissue was obtained by surgery from the primary tumor. From 14 patients tissue was taken by autopsy. From two patients tissue obtained by both surgery and later autopsy were available. From the autopsy patients tissue was taken only from the primary tumor (n = 6), from a metastasis (n = 1) and from the primary tumor and distant metastases (n = 7). Twelve of the tumors obtained by surgery were diploid, and two multiploid (two stem lines present). This was found both with image and flow cytometry. The group of patients could clearly be subdivided in short survivors (less than 9 months, n = 6) and long survivors (greater than 16 months, n = 8); since in both groups one multiploid and the remainder diploid cases were present, ploidy did not seem to be a good prognosticator for survival. In most (n = 26) of the tissues measured from the autopsy patients, again, a good correlation between image and flow DNA cytometry was obtained, the histograms being either (near) diploid or multiploid. In six cases, however, flow cytometry showed multiploidy whereas image showed aneuploidy (one single peak clearly deviating from diploidy). This discrepancy is caused because normal diploid (nonneoplastic) cells in the preparations could not be discarded from the flow cytometry measurements. Using the image cytometry data of the primary tumors, five diploid, three aneuploid, and four multiploid tumors were found. In five of the seven patients of whom tissue was obtained from the primary tumor and multiple metastases, differences between the histograms were found, mostly showing two malignant cell populations in one tissue and only one of them in another. Of one of the two patients of whom tissue was obtained by surgery and later autopsy, a change in histogram pattern was observed. It is concluded that although there is a high similarity between image and flow DNA cytometry, for an optimal interpretation of the histogram pattern, image measurements are more reliable. Ploidy determination does not seem to be of use in prediction of survival, and care should be taken in interpreting DNA histograms of metastases in SCCL patients because of the variability in histogram pattern.     

Measurement of cellular DNA content as an adjunct to diagnostic cytology in malignant effusions

We reviewed the final diagnosis and outcome of 119 patients who developed serous effusions. In addition to routine cytological examination, the cellular DNA content of fluid samples aspirated from the effusions was measured using flow cytometry in order to determine whether the detection of aneuploid cells could aid in diagnosis or serve as a guide to prognosis. The final diagnosis of 35 patients was non-malignant and a further 40 patients with biopsy-proven cancer had cytologically negative effusions. In all of these cases flow cytometry revealed the presence of diploid cells only. The effusions from 36 cancer patients were reported by cytology to contain a variable proportion of malignant cells, and aneuploid cells were detected in 23 of these samples, the remainder containing only diploid cells. Of 8 effusions where cytology was equivocal, one contained aneuploid cells and clinical outcome subsequently showed that all 8 were malignant. Median survival of patients with cancer was 3 months, and a positive cytology had no influence on survival. However, of the patients with positive cytology, those whose effusions contained aneuploid cells had a poorer short-term prognosis than those cases where only diploid cells could be detected (median survival 1.5 vs 4 months). Measurement of cellular DNA content using flow cytometry can occasionally confirm cancer in a cytologically equivocal effusion, but the negative results in 13 out of 36 (36.1%) effusions where cytology was reported as positive suggests that it has only a limited role in this clinical setting, using currently available techniques.     

Analysis of fibrinolytic proteins in relation to DNA ploidy in prostate cancer

The tissue concentrations of urokinase-type plasminogen activator (u-PA), urokinase-type plasminogen activator receptor (u-PAR), plasminogen activator inhibitor type 1 (PAI-1) and tissue-type plasminogen activator (t-PA) were investigated by an ELISA technique in normal and malignant samples of the prostate from 24 patients undergoing radical prostatectomy for organ-confined prostate cancer. The median concentration of u-PA was significantly higher in cancerous than in normal prostate tissue (p = 0.006). No significant increase of u-PAR, PAI-1 and t-PA was found in cancer tissue in comparison with the benign samples (p > 0.05). Assessment of the relationship between fibrinolytic proteins and DNA ploidy revealed an increased u-PA, u-PAR and PAI-1 in diploid prostate cancer as compared with the normal controls. However, in aneuploid cancer u-PA remained high but u-PAR and PAI-1 were decreased. This led to a higher local concentration of u-PA in aneuploid samples than in normal prostate and in diploid prostate cancer. No alteration of median t-PA was found in benign prostate or in diploid or aneuploid prostate cancer. The altered expression of u-PA, u-PAR and PAI-1 in diploid and aneuploid prostate cancer suggests a possible role of fibrinolytic proteins in the different biologic behavior of tumors, and may be one explanation for the higher metastatic potential of aneuploid tumors. Int. J. Cancer 78:320–325, 1998© 1998 Wiley-Liss, Inc.     

Characterization of bladder papilloma by two-parameter DNA-RNA flow cytometry

Two-parameter flow cytometry (FCM) studies of 0.9% NaCl solution bladder irrigation specimens were performed on 48 patients with histologically orderly or atypical papilloma of the urinary bladder in order to assess the value of RNA as a possible second parameter, along with DNA, in the detection of bladder tumors. DNA, RNA, and nuclear diameter measurements were obtained for each of 5000 cells/sample, and analyses were based on the distributions of those values. With the use of DNA content alone, 22 cases (46%) were classified positive by FCM. With RNA content as an additional parameter, 40 cases (83%) were positive. Two cases were suspicious, and 6 cases were normal by both parameters. Of 28 patients with papillomas showing histological atypia, 16 patients had positive DNA histograms, including 3 patients with aneuploid stemlines, but 24 of the 28 patients had positive RNA histograms. Of 20 patients with orderly papillomas, 6 patients had positive DNA histograms, including 3 patients with aneuploid DNA stem cell lines, but 16 of the 20 patients had positive RNA histograms. Thus, the probability of positive DNA histograms is higher in atypical papillomas (57%) than in orderly papillomas (30%), whereas elevated (positive) RNA is more characteristic of all papillomas without distinction between those that are histologically atypical (86% positive) or orderly (80% positive). For patients at risk of developing papillary bladder tumors, two-parameter DNA-RNA FCM appears to offer greater diagnostic sensitivity than does FCM based on DNA content alone.