DNA Flow Cytometric Study Research Articles

Design and Synthesis of Some New Furan-Based Derivatives and Evaluation of In Vitro Cytotoxic Activity.

New furan-based derivatives have been, designed, synthesized, and evaluated for their cytotoxic and tubulin polymerization inhibitory activities. DNA flow cytometric study of pyridine carbohydrazide 4 and N-phenyl triazinone 7 demonstrated G2/M phase cell cycle disruptions. Accumulation of cells in the pre-G1 phase and positive annexin V/PI staining, which may be caused by degeneration or fragmentation of the genetic components, suggested that cell death occurs via an apoptotic cascade. Furthermore, compounds 4 and 7 had a strong pro-apoptotic impact through inducing the intrinsic mitochondrial mechanism of apoptosis. This mechanistic route was verified by an ELISA experiment that indicated a considerable rise in the levels of p53 and Bax and a drop in the level of Bcl-2 when compared with the control.

Comparative DNA Flow Cytometric Study of Primary Intraocular and Central Nervous System Lymphomas

Primary intraocular lymphoma is generally considered as a subset of primary CNS lymphoma. This study attempts to show that they may in fact represent distinct entities by comparing their respective proliferation rates using DNA flow cytometry. Four samples of primary intraocular lymphoma and seven samples of primary CNS lymphoma were analyzed, all from paraffin-embedded tissue. All tumors were of the large B-cell type. A normal human tonsil sample was used as a control. Tissue samples were analyzed by DNA flow cytometry, which is a precise and objective method to measure DNA content and cell proliferation of a tumor. S-phase fraction (SPF) and DNA content were measured for each sample. The average SPF for primary intraocular lymphoma was significantly higher than that of primary CNS lymphoma, 23.8 (range: 18.9 to 29.6) versus 15.1 (range: 1.1 to 25.1) respectively. Of the 11 tumors analyzed, 2 brain tumors were aneuploid and 1 eye tumor was peridiploid. All other tumors were diploid. Thus, no significant pattern was detected in the DNA content of the tumors. This lack of clinical significance of tumor aneuploidy is consistent with data reported in the literature. The results of this study indicate that primary intraocular lymphoma is more aggressive and of higher grade than primary CNS lymphoma. The different proliferation rates of intraocular and CNS lymphomas may be explained by either their different spatial location or a distinct genetic composition, the latter reinforcing the hypothesis that the two are fundamentally different entities

DNA flow cytometric and interobserver study of crypt cell atypia in inflammatory bowel disease.

The pathological features and diagnostic reliability of crypt cell atypia (CCA) arising in inflammatory bowel disease (IBD) and its clinical significance are unknown. DNA flow cytometry (FCM) was performed on 14 colon biopsies of CCA from seven IBD patients (male-to-female ratio, 5:2; mean age, 53years; mean IBD duration, 15years) using paraffin-embedded tissue. Seven gastrointestinal pathologists were asked to diagnose each biopsy as negative for dysplasia (NEG), indefinite for dysplasia (IND), low-grade dysplasia (LGD) or high-grade dysplasia (HGD) by morphology alone, then again with knowledge of FCM results. Aneuploidy was detected in all 14 biopsies, and five of eight biopsies (63%) also showed strong and diffuse nuclear staining for p53 in the areas of CCA. Six (86%) patients developed HGD (n=5) or adenocarcinoma (n=1) in the same colonic segment where CCA had been diagnosed within a mean follow-up time of 27months. No follow-up information was available in the remainingone patient. When diagnoses were grouped as NEG or 'atypical' (including IND, LGD or HGD), the overall agreement rate of 76% (kappa=0.51) based on morphology alone improved to 90% (kappa=0.81) with knowledge of FCM results. Even when categorised as NEG or dysplasia (LGD or HGD) with each of the IND diagnoses reclassified into three categories (NEG, LGD or HGD) based on the degree of suspicion for dysplasia, the overall agreement rate of 63% (kappa=0.25) based on morphology alone improved to 73% (kappa=0.46) with knowledge of FCM results. However, when grouped as NEG, LGD or HGD, the overall agreement rate was less than 40% (kappa<0.09) regardless of knowledge of FCM results. The presence of aneuploidy, p53 positivity and development of HGD or adenocarcinoma on follow-up indicate that CCA likelyrepresents a dysplastic lesion (at least LGD) and is a histological marker of neoplastic progression. Although the grading of CCA, largely based on cytological abnormalities, is subject to significant interobserver variability, CCA can be histologically identified and should lead to a recommendation of increased endoscopic surveillance, especially if aneuploidy is detected.

Can we identify the group of small invasive (T1a,b) breast cancers with minimal risk of axillary lymph node involvement? A pathohistological and DNA flow cytometric study

The goal of this study was to identify a group of small (≤1 cm) breast cancers (T1a,b) with a particularly low probability of axillary lymph node metastases, where routine axillary staging may be unnecessary. We retrospectively analyzed 152 T1a,b breast carcinomas with axillary dissection surgically removed at Clinical Hospital Center Split (Croatia) in the period from 1997 to 2006. The analysis included 40 T1a,b cancers with, and 112 T1a,b cancers without axillary lymph node metastases. The basic morphological features of cancers were investigated histologically, while hormone receptors and HER2/neu were investigated immunohistochemically with an additional CISH analysis of HER2/neu 2+ cases. The ploidy and S-phase fraction were determined by DNA flow cytometry. The association of the investigated features with the likelihood of axillary lymph node metastases was analyzed by univariate and multivariate analysis. The univariate analysis showed that lymph node metastases were associated with tumor size (T1a/T1b; p = 0.026), histological type (ductal/non-ductal; p = 0.014), lymphovascular invasion ( p < 0.001), HER2/neu expression ( p = 0.04), ploidy ( p = 0.027), and combined values of ploidy and S-phase fraction ( p = 0.025). The lymphovascular invasion was the only independent factor associated with axillary nodal metastases ( p = 0.01). In the group of T1a,b cancers without lymphovascular invasion, HER2/neu expression ( p = 0.021) and combined values of ploidy and S-phase fraction ( p = 0.016) were independent factors associated with axillary lymph node metastases. This study showed that diploid T1a,b cancers with low S-phase fraction, which are also without lymphovascular invasion and HER2/neu amplification, represented the group of cancers with a low probability of axillary lymph node metastases.

The Clinical Value of Flow Cytometric DNA Content Analysis in Patientswith Soft Tissue Sarcomas

Purpose. The purpose of this study was to evaluate: (1) the correlation between grade and ploidy or S-phase fraction (SPF), (2) the prognostic value of DNA flow cytometric study in soft tissue sarcomas. Patients /Methods. In all, 47 tissue samples from soft tissue sarcoma patients, surgically treated in the same center, were included. Flow cytometric analyses were performed according to a modified version of the original method of Hedley et al. Results. DNA ploidy status could be determined in 44 samples out of 47 (success rate 94%). Of these 44, S-phase fraction could be calculated in 34 samples (77%). In the study group as a whole, aneuploidy was significantly correlated with high grade. Survival analyses were carried out in 21 patients with soft tissue sarcoma, all surgically treated in the same center, without chemotherapy or radiotherapy. In univariate analyses, DNA ploidy was found to be a significant factor for overall survival (OAS) and metastasis-free survival MFS. Mean OAS for aneuploid tumors and diploid tumors were 35 and 65 months (p=0.034), and mean MFS 23 and 61 months, respectively (p=0.005) . Discussion.There is a relation between histological grade and ploidy in soft tissue sarcomas. It appears that low-grade tumors are generally diploid, whereas high-grade tumors tend to be aneuploid. In a subgroup of patients treated only with surgery, DNA ploidy was found to be an important factor for predicting OAS and MFS.

Anorectal malignant melanoma. A clinicopathologic study, including immunohistochemistry and DNA flow cytometry.

Anorectal malignant melanoma is a rare tumor with an extremely poor prognosis. DNA flow cytometric study as well as detailed immunohistochemical study have not been reported previously. Eighteen cases of anorectal melanoma were studied, including immunohistology for melanoma markers and epithelial markers and DNA flow cytometric study of paraffin blocks. Most patients were Ashkenazi Jews, compared with Sephardi Jews and Arabs. Of the 17 patients followed, 14 died of disease at 4-39 months from presentation. Three patients were alive with disease at 12, 53, and 72 months of follow-up. Tumor thickness ranged from 3-35 mm (mean, 12.8 mm). The 2 long term survivors had tumor thickness < or = 7 mm. No correlation was found between the mode of primary surgical treatment (8 patients: abdominoperineal resection; 10 patients: local excision) and outcome. Vimentin, HMB-45, and S-100 protein stainings were positive in 18, 17, and 15 tumors, respectively. Polyclonal carcinoembryonic antigen (CEA), broad-spectrum cytokeratin, epithelial membrane antigen, monoclonal CEA, and TAG-72 (B72.3) stainings were positive in 13, 3 (only focal and rare staining), 2, 0, and 0 tumors, respectively. Thirteen tumors had adequate material for DNA analysis, and all were DNA aneuploid. S-phase fraction could be assessed in 11 tumors and ranged from 7.7-24% (mean, 14%). An S-phase fraction of < 10% was observed in the 2 long term survivors. Anorectal melanoma in this study carried a grave prognosis. The frequent staining for polyclonal CEA (with negative monoclonal CEA staining) was probably due to nonspecific cross-reacting antigens. The occasional staining for epithelial markers warrants a comprehensive immunohistochemical study to ensure a correct diagnosis, especially in small biopsies of amelanotic undifferentiated tumors that lack junctional changes. The aneuploidy of all tested tumors reflected their highly malignant behavior. A trend toward longer survival was observed in patients with thin tumors and an S-phase fraction of < 10%. However, due to the small number of survivors, the latter observation should be further tested in a larger scale series.

Chromophobe cell renal carcinoma: DNA flow cytometry and proliferation status in 9 cases.

Nine cases of chromophobe cell renal carcinoma are reported, 2 in pregnant women. One neoplasm progressed and the patient died of disease, and this tumor presented the highest mitotic index and focal positivity to Ki-67 antibody. DNA flow-cytometric study discovered 89% of aneuploidy, with 3 tetraploid cases. We conclude that chromophobe cell renal carcinoma is a relatively indolent but clearly malignant neoplasm. We suggest that the mitotic index could be the best method to predict malignant behavior. The high incidence of aneuploid cell lines may be due to structural anomalies; it is not indicative of malignant behavior.

Prognostic factors in carcinoma of the vulva: a clinicopathologic and DNA flow cytometric study.

The clinical staging of carcinoma of the vulva is a predictor of patient survival; however, the significance of other prognostic factors remains somewhat controversial. Length of survival after diagnosis of invasive squamous cell carcinoma was determined for 39 clinically staged and surgically treated patients who were followed at our institution. Clinical stage, tumor type, use of radiotherapy (RT), histopathologic features (invasive pattern, depth of invasion, lymph node status, nuclear grade, adjacent dysplasia, desmoplasia, inflammation) and DNA ploidy (determined by flow cytometry from paraffin-embedded tissue) were evaluated as predictors of survival. Kaplan-Meier survival curves were generated for strata defined by each of the various predictors and compared using the log-rank test. Advanced stage (p = 0.0002), RT use (p = 0.0004), "spray" invasive pattern (p = 0.005), positive lymph node status (p = 0.001), increased positive lymph node number (p = 0.016), and greater depth of invasion (p = 0.039) were associated univariantly with decreased survival time. Spray invasive pattern (p = 0.018), positive lymph node status (p = 0.030), positive lymph node number (p = 0.040), and RT use (p = 0.045) continued to be associated with decreased survival time after controlling for stage. Of the significant factors, invasive pattern stands out as a qualitative feature that may have potential benefit in predicting survival independent of clinical stage in patients with vulvar carcinoma.

Chondrosarcoma of bone. A clinical and DNA flow cytometric study

Fifty-two patients with chondrosarcoma were treated in our hospital between 1981 and 1992. Tumours from 21 of the patients were histologically grade I, 19 were grade II, 10 were grade III and two were grade IV. Forty-seven patients were treated surgically and samples for flow cytometric DNA analysis were taken from 38 tumours with a successful measurement in 37 cases. The mean follow-up time was 40 months and during this period 17 of the patients treated surgically developed a local recurrence or metastasis. In one case, a highly anaplastic sarcoma developed after radiation treatment. Only two of the 19 diploid tumours recurred, whereas eight of the 18 aneuploid tumours showed a recurrence. These results indicate that DNA flow cytometry is of prognostic significance in chondrosarcoma and may aid in planning the treatment.

FLOW CYTOMETRIC CELLULAR DNA CONTENT AND LYMPH-NODE METASTASIS IN SQUAMOUS-CELL CARCINOMA OF THE ORAL CAVITY

This prospective DNA flow cytometric study on 386 primary squamous cell carcinomas of the oral cavity showed that only 18% of the patients with diploid primary tumors had lymph node metastasis on admission compared to 52% of those with aneuploid carcinomas. The aneuploid group without evidence of lymph node involvement at the time of primary tumor treatment carried a 3-fold increased risk for developing late metastasis (23%) compared with the diploid group (8%). The clinical manifestation of occult metastasis in patients with diploid carcinomas was delayed by about two years compared to the aneuploid group. These ploidy-specific differences of the metastatic behaviour held true even if stratified with respect to tumor stage, histological grade and tumor localization. These results provide substantial evidence that cells with gross DNA content aberrations have a significantly higher probability of successfully producing a metastatic colony than flow cytometrically diploid tumor cells. An excellent 5-year survival rate of 90% in the diploid NO group in contrast to 52% in aneuploid NO cases and an even worse survival rate of 21% in patients with lymph node involvement at presentation underline the clinical importance of these findings.

Peripheral primitive neuroectodermal tumour and extra-osseous Ewing's sarcoma; a histological, immunohistochemical and DNA flow cytometric study

Although peripheral primitive neuroectodermal tumour (pPNET) and extra-osseous Ewing's sarcoma (EES) are thought to be closely related neoplasms, their clinical behaviour differs considerably. To determine the clinical relevance of the Schmidt classification scheme for differentiating pPNET and EES, 20 tumour specimens of poorly differentiated round cell tumours were evaluated. In addition, the diagnostic value of several neural markers and the prognostic value of quantitative morphological variables (DNA ploidy, S-phase fraction, and the mitotic activity) were assessed. Homer-Wright rosettes were present in 9 tumours. Neuron specific enolase (NSE) was expressed in 11 tumours, 8 of which expressed a second neural marker (CD57, S100, or neurofilament). According to the Schmidt classification, 11 pPNET and 5 EES were distinguished. HBA-71 was exclusively expressed in pPNET and EES. The remaining tumours were classified as sarcoma not otherwise specified (n = 2), rhabdomyosarcoma (n = 1), and desmoplastic tumour with divergent differentiation (n = 1). EES611 patients fared significantly better than the pPNET patients (100% versus 42% 5-year survival). Neither DNA ploidy nor S-phase fraction assessed in 12 evaluative histograms (9 pPNET and 3 EES), nor mitotic activity yielded information of additional prognostic value. On the basis of this study and the Schmidt classification scheme, it can be concluded that if the diagnosis of EES and pPNET is based on light microscopy (Homer-Wright rosettes) and/or immunohistochemistry (at least two neural markers, i.e. NSE, S-100, CD57, and neurofilament), the classification provides important clinical information. Furthermore, positivity for HBA-71 is helpful in differentiating pPNET and EES from all other small round cell tumours.

CYTOGENETIC PROGRESSION AND PROGNOSIS IN ORAL-CARCINOMA - A DNA FLOW CYTOMETRIC STUDY ON 317 CASES

A total number of 317 consecutive patients with squamous cell carcinoma of the oral cavity were analysed by DNA flow cytometry. The proportion of tumors consisting exclusively of diploid cells decreased from 75% in T1G1 carcinomas to less than 10% in advanced tumors in favor of cases characterized by the presence of cell lines with aberrant DNA content. This observation indicates a genetic evolution of tumor clones with severe karyotype aberrations from diploid progenitors which obviously represents an ultimate event in the pathogenesis of oral carcinomas. Aneuploidy formation clearly contributes to the acquisition of progressive malignant behaviour as is underlined by a dramatic difference of the survival outcome of patients with diploid tumors (5-year overall survival: 88%) as compared to those who had aneuploid tumors (36%; P<0.001), a relationship which held true even if identical tumor stages were considered. There were no indications of successive genetic changes of the cellular DNA contents during tumor progression which would explain the wide range of variation of the individual DNA values. This observation supports the hypothesis of an aneuploidy formation by a single cytogenetic event, such as an abnormal mitosis. Although there were indications of an even worse survival outcome in the group of patients with peri-diploid tumor cell lines (P<0.1), a significant influence of different DNA contents on the prognosis could not be detected.

Primary carcinoid tumor of testis. Immunohistochemical, ultrastructural, and DNA flow cytometric study of three cases with a review of the literature

The cases of three patients with primary carcinoid tumor of the testis were reported. The patients were 41, 44, and 83 years of age. At initial examination, all three had testicular masses with or without associated pain, and none had the carcinoid syndrome. The tumors measured 4.3 cm, 3.0 cm, and 6.5 cm in dimension. All three tumors manifested classic histologic features of carcinoid tumors. The neoplastic cells exhibited argyrophilia, and all were immunoreactive to chromogranin, serotonin, neuron-specific enolase, and cytokeratin. Two tumors had positive test results for gastrin and one had positive test results for substance P and vasoactive intestinal polypeptide. No tumors reacted with somatostatin, insulin, pancreatic polypeptide, or placental alkaline phosphatase. Intracytoplasmic, membrane-bound, round-to-elliptical pleomorphic granules were identified by ultrastructural analysis in all cases. DNA flow cytometric analysis revealed a low degree (near-diploid) DNA aneuploidy in all cases, with a DNA index of 1.15 in two tumors and 1.3 in the third tumor. The three patients are alive and well 11 years, 7 years, and 6 months, respectively, after diagnosis. A total of 57 cases of this entity, including the 3 reported here, have been reported. Of these, 43 were pure carcinoid, and 14 were associated with teratoma; 6 (11.6%) patients developed metastases. Tumor size and the presence of carcinoid syndrome have been found to correlate with metastatic potential. Neither tumor necrosis nor local tumor invasion (into vessels, tunica albuginea, etc.) correlated with adverse prognosis. Carcinoid tumor of the testis is a rare indolent neoplasm with potential for distant metastases.

DNA flow cytometric study of thymic epithelial tumors with evaluation of its usefulness in the pathologic classification

A DNA flow cytometric study was performed on 64 thymic epithelial tumors using the modified Hedley method with formalin-fixed, paraffin-embedded tissues to evaluate whether ploidy analysis can be a useful aid in differentiating invasive thymoma (IT) from noninvasive thymoma (NT) and in understanding the ploidy pattern of various histologic types of thymic carcinomas (TCs). The IT group was further subdivided into macroinvasive thymoma (macro-IT) and microinvasive thymoma (micro-IT). Six cases were excluded due to a coefficient of variation greater than 6. Aneuploidy with a DNA index greater than 1.05 was found in every group: two of 16 NTs, two of nine micro-ITs, 11 of 13 macro-ITs, and 12 of 20 TCs were aneuploid. However, the proportions of aneuploidy of macro-IT and TC were statistically significantly higher than those of NT and micro-IT ( P < .001). Similarly, the DNA indexes of macro-IT and TC were significantly higher than those of NT and micro-IT ( P < .004). Therefore, micro-ITs were cytometrically similar to NTs and could be grouped with the NTs. Although ploidy analysis cannot predict whether an individual thymoma is invasive, aneuploidy and a high DNA index would favor a macro-IT, which also has a significantly higher S-phase fraction. Thymic carcinoma and IT cannot be differentiated by DNA flow cytometry. No definite ploidy pattern was observed for various histologic types of TC, but transition from diploid to aneuploid was demonstrated in thymomas undergoing malignant transformation into TC.

DNA Heterogeneity in prostatic adenocarcinoma. A DNA flow cytometric mapping study with whole organ sections of prostate

The degree of DNA heterogeneity varies between tumors arising at different sites. The presence of a significant degree of variability within a given tumor may result in problems in the interpretation of DNA flow cytometric findings. This study evaluated the degree of DNA heterogeneity in prostatic adenocarcinoma. A total of 122 3-mm punch biopsy specimens were evaluated from single representative whole organ sections from nine cases of prostate cancer (range, 4-18 samples per case; mean, 12 samples; median, 14 samples). Individual punch biopsy specimens were graded and reviewed to confirm the presence of carcinoma and processed for DNA ploidy analysis. Assessable histograms, defined as having a coefficient of variation of the diploid G0/G1 peak of 7.5% or less, were available for 111 (91%) of the samples. Of the nine cases studied, five (56%) showed heterogeneity in the DNA pattern (diploid plus aneuploid, n = 1; diploid plus tetraploid, n = 2; and diploid plus tetraploid plus aneuploid, n = 2). All four cases having a homogeneous DNA content were DNA diploid in all samples. In those cases with a heterogeneous pattern, the areas having abnormal DNA patterns could not be predicted by histologic pattern or grade. From this study, the authors conclude that a significant degree of DNA heterogeneity exists within individual cases of prostatic adenocarcinoma, and this may be an important confounding factor in DNA ploidy studies.

The expression of proliferating cell nuclear antigen in paraffin sections of peripheral, node-negative non-small cell lung cancer.

Cell proliferation of 40 peripheral, node-negative non-small cell lung cancers (NSCLC) treated with surgery alone was investigated by immunohistochemical analysis with the monoclonal antibody (MoAb) PC10, which recognizes a proliferating cell nuclear antigen (PCNA) in formalin-fixed and paraffin-embedded material. Results were correlated with DNA ploidy and S-phase fraction (SPF) analyzed by DNA flow cytometric study. Mitotic count (MC) was analyzed by light microscopic study and histopathologic features. PCNA immunoreactivity was seen in all samples and confined to the nuclei of cancer, but not to the surrounding, tumor-negative cells; its frequency ranged from 0-70% (median, 15%), and tumors expressed either a low (0-25%, n = 25) or intermediate (26-75%, n = 15) proliferative activity. There was no relationship between PCNA immunoreactivity and tumor stage or among size, histologic type, and mitotic count (MC). Tumors with intratumoral blood vessel invasion (BVI) showed a significantly higher (P less than 0.005) PCNA immunoreactivity than BVI-negative tumors. PCNA scores were significantly higher (P less than 0.005) in DNA aneuploid (n = 22) than in DNA diploid (n = 18) tumors and correlated significantly with the SPF of DNA aneuploid tumors (r = 0.825, P less than 0.0001), but not with diploid tumors (r = 0.002, P = 0.9). Intermediate proliferating tumors had a significantly higher (P less than 0.01) MC than their counterparts. In univariate analysis, significant predictors of survival were tumor classification (T1 versus T2), tumor size (less than or equal to 2.6 cm versus more than 2.6 cm), BVI (BVI-negative versus BVI-positive), MC (less than or equal to 8 versus more than 8), and PCNA immunoreactivity (low versus intermediate). DNA ploidy and SPF did not influence survival significantly. Only PCNA immunoreactivity retained its independent level of significance (P = 0.02) by multivariate analysis. It was concluded that PCNA immunostaining is a simple and clinically useful method for estimating cell proliferation in formalin-fixed, paraffin-embedded tissue of resected peripheral, node-negative NSCLC.

A flow cytometric study of non-small cell lung cancer classified as T1N0.

Surgical resection currently offers the best chance for cure of non-small cell lung cancer but its efficacy is limited by subsequent tumor recurrence. Even the most favorable cancers (T1N0 tumors) recur 20% to 30% of the time within 5 years and there is currently no way to anticipate precisely which tumors will recur. To test whether DNA flow cytometric study might be useful in this regard, the authors performed a retrospective case-control study of 102 tumors (51 recurrent cases and 51 controls) from a prospective registry of patients with completely resected, meticulously staged T1N0 non-small cell carcinomas. Unbiased relative hazard ratios of recurrence were estimated for ploidy and proliferative rate, as well as for tumor histologic type and clinical variables. Ploidy abnormalities were slightly more common among cases (67%) than controls (57%) but this difference was not statistically significant. Estimation of proliferative rates was possible for 85 tumors but there was no significant difference between cases and controls and proliferative rates were not prognostic of recurrence. In multivariate analyses, the observed predictive value for each of the flow cytometric parameters was modest at best and smaller than that seen for tumor histologic type. These results suggest that flow cytometric analysis has limited value in guiding management of patients with early stage non-small cell carcinoma.

Simultaneous carcinoma involving the endometrium and the ovary. A clinicopathologic, immunohistochemical, and DNA flow cytometric study of 18 cases

Eighteen carcinomas involving both the endometrium and the ovary were studied. Stage, size, bilaterality and pattern of ovarian involvement, histologic types and grades, presence of endometrial hyperplasia or ovarian endometriosis, myometrial, tubal, lymphatic and blood vessel invasion, and follow-up of the patients were all evaluated. Accordingly, the cases were classified as follows: Group A (nine cases), two separate primary tumors; and Group B (nine cases), uterine primaries with ovarian metastasis or ovarian primaries with uterine metastasis. Immunohistochemical stains (CAM 5.2, wide-spectrum keratin, vimentin, carcinoembryonic antigen (CEA), CA 12.5, CA 19.9) were performed in all cases. Flow cytometric determination of nuclear DNA was done in 13 (seven Group A and six Group B tumors). Of the nine cases with independent primary tumors, seven showed different immunohistochemical profiles in the ovarian and uterine tumors, whereas only four of the nine metastatic ones had similar staining characteristics. Five cases with independent primary tumors, but only one of the metastatic group, exhibited different aneuploid stemlines in the endometrial and ovarian tumors. The other seven (two independent and five metastatic) cases had similar DNA indexes in both tumors. Immunohistochemical and DNA flow cytometric study may be of some value for the distinction between metastatic and independent tumors, but differential diagnosis must presently rely largely upon conventional clinicopathologic criteria.

Simultaneous renal and transitionalcell carcinomas in same kidney with subsequent lymph node metastasis: DNA flow cytometric study

We report the DNA content in simultaneous renal and papillary transitional cell carcinomas in the same kidney with a subsequent metastasis of the transitional cell carcinoma to a left supraclavicular lymph node. The flow cytometric DNA analysis revealed aneuploid stemlines with identical DNA indices in both the transitional cell carcinoma and the metastatic lesion and a diploid pattern in the renal cell carcinoma. A discussion of these findings and the potential implication of DNA ploidy analysis in similar situations is presented.

Epithelioid pleural mesotheliomas and pulmonary adenocarcinomas: A comparative DNA flow cytometric study

The DNA flow cytometric characteristics of 23 immunohistochemically and ultrastructurally proven pleural epithelioid mesotheliomas were compared with those of 41 primary pulmonary adenocarcinomas. Multiple, separate tissue blocks were analyzed from each neoplasm to assess DNA heterogeneity. All of the pleural mesotheliomas and 80% of the pulmonary adenocarcinomas manifested a homogeneous DNA ploidy (DNA stability). A significant statistical difference in the ploidy pattern between pleural mesotheliomas and pulmonary adenocarcinomas was noted ( P < .001). Mesotheliomas were mostly diploid (78%) and adenocarcinomas were preponderantly aneuploid (88%). The proliferative rate and DNA indexes of the aneuploid adenocarcinomas were significantly higher than those of pleural mesotheliomas ( P < .001). There was no statistical significance between the proliferative rates of diploid mesotheliomas and those of adenocarcinomas. We conclude that the DNA flow cytometric characteristics of mesotheliomas are significantly different from those of pulmonary adenocarcinomas. The clinical implications of these findings are discussed.