Incidence Of DNA Aneuploidy Research Articles

High frequency of DNA aneuploidy detected by DNA flow cytometry in Bowen’s disease

To detect DNA aneuploidy in Bowen’s disease, we investigated DNA flow cytometric analysis. Single cell suspensions were prepared from 18 fresh samples histopathologically diagnosed as solitary Bowen’s disease and analyzed by DNA flow cytometry. In 16 (89%) of 18 lesions, DNA aneuploidy was demonstrated with a single aneuploid peak. DNA indices ranged from 1.29 to 1.74. The incidence of DNA aneuploidy in Bowen’s disease is higher than those of cutaneous squamous cell carcinoma, which was 25–80% in the previous reports. Therefore, in Bowen’s disease, DNA aneuploidy may not imply a good marker for characteristics of non-melanoma skin cancer. A single aneuploid peak commonly observed in Bowen’s disease suggests that this disease consists of the monoclonal proliferation of keratinocytes containing abnormal DNA content.

The incidence of DNA aneuploidy in multiple myeloma does not correlate with stage of disease.

The bone marrow of 47 patients with multiple myeloma (MM), 2 patients with plasma cell leukemia, and 11 patients with monoclonal gammopathy of undetermined significance (MGUS) was analyzed by flow cytometry for the detection of DNA aneuploidy. The question to be addressed was whether a correlation exists between the incidence of DNA aneuploidy and the stage of disease. With one-parameter analysis of DNA staining, a DNA aneuploidy was detectable in 17 of 60 patients. The detection rate for DNA aneuploidies could be increased to 37 of 60 patients with the second method, the simultaneous measurement of DNA content in additional immunophenotyped cells (CD38 or B-B4). In MGUS and the early stages of MM, the discrepancy between both methods was higher than in stage III MM. There were no statistical differences in the incidence of DNA aneuploidy between MGUS or the early stages of MM and stage III MM (21/32 patients vs 16/26 patients). The CD56 expression in plasma cells was significantly higher in cases of DNA aneuploidy (mean +/- SD, 64.7% +/- 33.65% vs 39.3% +/- 36.69%; P = .028). Comparing the ratio of diploid to DNA aneuploid with that of CD56+ to CD56- plasma cells, a correlation was found in MGUS (r = 0.7320) in the early stages of MM, I and II (r = 0.8023), but not in stage III MM. Based on these data, the dual-staining method for DNA content in immunophenotyped cells is preferred for the detection of DNA aneuploidy, especially in the early stages of MM and in MGUS. The clinical importance of a classification of DNA aneuploidy and CD56 antigen expression together is proposed for testing.

Cell-proliferative activity and its relationship to the histologic type of early gastric cancer.

To evaluate the biological characteristics of gastric carcinoma and of the normal gastric mucosa from which carcinoma seems to develop, we analyzed the DNA content and proliferative activity of cells in 154 patients with primary mucosal gastric cancer and 40 patients with gastric ulcer by flow cytometric analysis. A total of 154 samples from mucosal gastric carcinomas, 154 samples from normal gastric mucosa adjacent to carcinomas and 40 samples from normal gastric mucosa adjacent to gastric ulcers were subjected to analysis. The incidence of DNA aneuploidy of differentiated carcinoma cells (34.3%) was significantly higher than that of undifferentiated carcinoma cells (14.5%, P=0.014). Among the cancers with diploid DNA, the mean value of the proportion of cells in the S phase of the cell cycle in differentiated carcinomas was 6.64% and it was significantly higher than that in undifferentiated carcinomas (5.53%, P=0.01). The mean values for the S and G2 + M phases for cells in the normal gastric mucosa from sites adjacent to the differentiated carcinomas were 5.29% and 1.81%. These values were higher than those for cells in normal gastric mucosa from sites adjacent to undifferentiated carcinomas (4.89% and 1.40%, respectively) and than those for cells in normal mucosa from sites adjacent to gastric ulcers (4.48% and 1.68%, respectively). During the early phase of gastric cancer, differentiated carcinoma seems to develop from gastric mucosa where cells have high proliferative activity. Reflecting this biological characteristic, the cells of differentiated carcinomas also have high proliferative activity.

Flow cytometric DNA content analysis of ultraviolet light-induced squamous cell carcinomas: a comparative study of squamous cell carcinomas of the lip and those arising from other sites of sun-damaged skin.

The development of squamous cell carcinoma (SCC) of the lip is considered to be mainly related to excessive sun exposure. However, its higher metastatic rate is distinct from that of SCCs arising from other sites of sun-damaged skin. Flow cytometric DNA content analysis, using paraffin-embedded specimens, was performed on 15 SCCs of the lip and 32 SCCs arising from other sites of sun-damaged skin. A significantly lower incidence of DNA-aneuploidy was observed in SCCs of the lip (2/15) than in those of sun-damaged skin (15/32) (p < 0.05). The mean age of patients with SCC of the lip (66.7 +/- 11.6 years; mean +/- SD) was significantly lower than that of the other patients (78.1 +/- 11.1 years) (p < 0.01). There was no significant difference between the mean diameter size of tumors on the lip (19.5 +/- 5.7 mm) and that of tumors on other sites of sun-damaged skin (30.7 +/- 20.5 mm). These results suggest that additional carcinogenic factors besides ultraviolet light may be involved in the development of SCC of the lip.

DNA flow cytometry in gastric carcinoma: implication in patients with potentially curative resection.

The relevance of DNA ploidy as a prognostic factor in patients with gastric cancer is controversial. The prognostic significance of DNA ploidy and its relationship to conventional histological grading and staging of the tumor (TNM stage, Lauren, Ming and WHO classification) were evaluated. DNA ploidy of the tumor was determined by flow cytometry on archival material from 76 patients who underwent R0, D2 stomach resection. DNA aneuploidy was found in 39 cases (51%). No significant association between DNA aneuploidy and either patients' sex, pT, pN, type according to Ming or Borrmann and tumor localization was found. The incidence of DNA aneuploidy was significantly lower in tumors of diffuse type according to Lauren, in signet-ring cell or undifferentiated type (WHO), in grade 3/4 tumors, and in patients younger than 50 years. We found no significant difference in survival of patients with DNA aneuploid when compared to DNA diploid tumors, although the prognosis of the patients with lower DNA index (DI < 1.2) tended to be better than that of higher DNA index (DI > 1.2). DNA ploidy appears to be of limited prognostic value after R0, D2 resection of stomach cancer.

DNA aneuploidy in acute myeloblastic leukemia is associated with a high expression of lymphoid markers.

In the present study the DNA cell content of 205 de novo acute myeloblastic leukemia (AML) patients is analyzed at flow cytometry in order to determine both the incidence of DNA aneuploidy in AML patients and the clinical and biological characteristics of AML aneuploid cases. All technical procedures were performed in accordance with the proposed guidelines of the DNA Cytometric Consensus Conference. Our results show that the incidence of DNA aneuploidy is quite low (4.8%), with most cases (n = 8) hyperdiploid and only a small proportion (n = 2) hypodiploid. No major differences were detected between the aneuploid and diploid cases with respect to the clinical and prognostic disease characteristics. Regarding the immunophenotype of the blast cells, the aneuploid cases displayed both a higher incidence of immature myeloblastic phenotype and a greater expression of lymphoid-associated antigens. In summary, our results show that AML patients display a quite low incidence of DNA aneuploidy and that despite the fact that these cases do not display particular clinical characteristics, they show an association with the expression of lymphoid-related markers.

DNA-PLOIDY ANALYSIS WITHIN SELECTED REGIONS OF COLORECTAL ADENOMAS CONTAINING CARCINOMA

In order to better understand the relationship of DNA ploidy, dysplasia, early cancer, and colorectal tumor progression, 11 colorectal adenomas containing carcinoma invading the submucosa were investigated using DNA flow cytometry. Multiple frozen samples were taken from the selected sectors corresponding to adenoma tissue with low-grade dysplasia, high grade dysplasia and early cancer. Sampling accuracy was performed under histologic examination by multiple cryostatic sections. Data were compared with previously reported results in non-cancerous adenomas and advanced carcinomas. Incidence of DNA aneuploidy among the dysplastic regions of the adenomas containing carcinomas resulted higher than that observed in non-cancerous adenomas (p=0.02). Furthermore, among the DNA aneuploid populations, the frequency of clones with high DNA Index (DI>1.3) was slightly higher in adenomas with cancer than in adenomas without cancer (p=0.07). We suggest that differences may exist in DNA aneuploidy evolution between these two types of lesions. In early cancer, the near-diploid clones were 57% with respect to 18% (p=0.01) in advanced cancer since in this latter case the majority of the DNA abnormal clones were in the near-hypertriploid region (82%). Thus, the acquisition of the invasive phenotype appears to be linked with the expansion and stabilization of high DNA aneuploid clones. Further analysis on a larger number of cases of adenomas containing carcinoma are necessary to validate these interpretations.

Evaluation of cytophotometric DNA content abnormalities in premalignant and preinvasive breast lesions

Progression of breast neoplasia is characterized by a variety of causal and nonspecific molecular, karyotypic, and cellular level genetic alterations. These include allelic losses, chromosomal rearrangements, and aneusomies, as well as widely divergent clonal DNA content aberrations. Establishment of the sequence and the pathologic significance of individual changes has been challenging owing to disease heterogeneity, protracted natural history, and difficulties in sampling and localizing precursor lesions. These limitations have driven attempts at cytophotometric evaluation of premalignant/preinvasive proliferations using histologic (i.e., in situ) microdissection of intact paraffin-embedded tissue sections. Using image cytophotometric measurements in preserved sections, clonal DNA content abnormalities are identified in up to three-fourths of preinvasive breast carcinomas. The incidence of DNA aneuploidy is grade-dependent and similar in degree to invasive lesions. Comparison of ploidy determinations between preinvasive and corresponding invading populations, however, suggests host tissue permeation is accompanied by measurable DNA content shifts in many cases. Image cytophotometric DNA content abnormalities are also detectable in florid/atypical proliferative lesions, albeit less frequently (∼25% of cases) and to a lesser extent (i.e., near diploid) than cytologically malignant lesions. Although the sensitivity of cytophotometric ploidy assessments in tissue sections is limited by nuclear sectioning, the presence of genomic instability in premalignant lesions is supported by evidence of individual chromosome aneuploidy, demonstrated by interphase cytogenetics with fluorescent centromere-specific probes. These data suggest cytophotometrically detectable DNA content anomalies may precede unequivocal morphologic transformation in breast neoplasia. Further, clonal DNA content heterogeneity in breast carcinoma may accompany biologically critical steps in neoplastic progression of breast tumors.

Characteristics and problems of flow cytometric nuclear DNA content analysis in lung cancer using bronchoscopically-obtained specimens

Flow cytometric nuclear DNA content analysis was performed on 106 bronchoscopically-obtained specimens from 67 patients with primary lung cancer. Brushing, curettage and biopsy samples in which tumor cells were identified by microscopic examination were considered suitable for analysis. The incidence of DNA aneuploidy in small cell carcinoma (77.8%) was higher than that in any other histological type of lung cancer. Intratumoral heterogeneity was found to affect the detectability of DNA aneuploidy, suggesting that care must be taken to obtain adequate samples of tumor cells and to consider intratumoral heterogeneity. These results indicate that this method can be used for nuclear DNA content analysis not only in cases of resectable lung cancer but also for unresectable lung cancers and may provide useful biological information in the clinical management of all types of lung cancer.

Intratumoral heterogeneity in DNA ploidy of bladder carcinomas.

In order to investigate whether DNA ploidy in a single area of a tumor could be representative of the entire tumor, flow-cytometric DNA analysis was performed for multiple samples (an average of 7 samples) obtained from the same tumor in 9 cases of transitional cell carcinoma of the urinary bladder. The incidence of DNA aneuploidy was 88.9% (8 out of 9 cases) and, furthermore, in 7 of these, intratumoral regional heterogeneity in DNA ploidy was evident. With high frequency, bladder carcinoma consisted of heterogeneous cell populations with different ploidy. There were 2 cases in which the tumor consisted of mixed subpopulations of diploid and aneuploid clones which were located in different parts of the same tumor. In such cases, an aneuploid line could not have been detected in a single specimen from the tumors.

Intratumoral heterogeneity in DNA ploidy of esophageal squamous cell carcinomas

Intratumoral heterogeneity in DNA ploidy was investigated in 23 cases of squamous cell carcinoma of the esophagus. Nuclear DNA content was determined for multiple samples taken from the same tumor, using a flow cytometric technique. The incidence of DNA aneuploidy was 87% in this series, and DNA indices ranged from 0.78 to 2.64 but most of them fell within values between 1 and 2. Of these cases ten (43.5%) showed intratumoral heterogeneity in DNA ploidy; in addition to a diploid population, one to four heterogeneous aneuploid subpopulations were discernible in the same tumor. However, morphologic variation was minimal within the same tumor. DNA index seen in metastatic lesions was identical with one of those in the primary lesion. Mechanisms responsible for intratumoral difference in DNA ploidy are also discussed from the aspect of tumor progression.

Human immunodeficiency virus-related lymphomas: a possible association between tumor proliferation, lack of ploidy anomalies, and immune deficiency.

In an attempt to identify a biologic basis for the aggressive clinical behavior of human immunodeficiency virus (HIV)-associated lymphomas (HAL), dual-parameter flow-cytometric analysis was performed on 22 paraffin-embedded biopsy specimens. Cases were analyzed for DNA ploidy, the percentage of cells in S-phase (proliferative activity), and content of a recently identified proliferation-associated nuclear antigen, p105. The DNA-content analysis of 22 HALs was compared with that of 109 cases of intermediate-grade non-Hodgkin's lymphoma (NHL) unrelated to the acquired immune deficiency syndrome (AIDS) studied previously in our laboratory and 125 cases of high-grade NHL reported in the literature. The proliferative activity was higher in intermediate-grade HAL relative to non-AIDS NHL (24.0% v 10.4%; P = .03), and in high-grade HAL in comparison with NHLs of similar histology unassociated with HIV infection (24.8% v 19%), although the latter did not reach statistical significance. The number of mitoses per 10 high-power fields was found to correlate with the percentage of cells in S-phase (r = .68; P = .0004). Although p105 content tended to be higher in HAL than in an AIDS-related complex (ARC)-associated hyperplastic lymph node control, no statistically significant associations were found between p105 content and proliferative activity or the number of mitoses per 10 high-power fields. When compared with non-AIDS NHLs of comparable grade, there was a trend toward a lower incidence of DNA aneuploidy in both intermediate- (25% v 56%) and high-grade (38.5% v 60%) HALs. The higher proliferative activity and lower incidence of DNA aneuploidy found in HAL relative to non-AIDS NHL of comparable histologic grade may represent differences in pathogenesis and may underlie the poor prognosis of HIV-associated NHL.

DNA Ploidy Patterns in Gallbladder Adenocarcinoma

DNA ploidy patterns in gallbladder carcinoma were examined by flow cytometry using paraffin-embedded tissues. The incidence of DNA aneuploidy was 46.3% and of a diploid pattern, 53.7%. No relation could be observed between DNA ploidy pattern and histological type. There was a tendency for the incidence of diploid patterns in early carcinomas to be higher than that of aneuploidy patterns, but there was no significant difference. No obvious correlation could be demonstrated between DNA ploidy pattern and prognosis. Gallbladder carcinomas were divided into metaplastic and non-metaplastic types based on the presence or absence of metaplastic changes, in accordance with our classification. The relation between DNA ploidy pattern and tumor type was examined, but none was observed. In the present study, no correlation could be observed between DNA ploidy pattern and clinicopathologic findings in gallbladder carcinoma.

Flow Cytometric DNA Measurement of Gastric Cancers: Clinico-Pathological Implication of DNA Ploidy

The relationship of DNA ploidy to clinical, pathological or prognostic data was investigated in 70 surgically removed gastric cancers, 60 of which were advanced cases. The cellular DNA content was measured by means of flow cytometry. Overall, DNA aneuploidy was seen in 43 of the cases (61%) while the normal gastric mucosa consisted of only a diploid line without any aneuploid population. Ploidy levels of gastric cancer spanned a wide range from a hypodiploid value to a hyperpentaploid value, although near-diploid and near-triploid cases were the majority in the series. No significant association between ploidy and either age or sex was found. The incidence of DNA aneuploidy was significantly higher in differentiated adenocarcinomas than in undifferentiated ones (p less than 0.05). However, DNA ploidy was not related to some pathologic characteristics such as the extent of tumor invasion and the frequency of lymph node metastasis. According to Kaplan-Meier survival plots, prognosis of the patients with DNA diploid tumors tended to be better than that of aneuploid tumors, but the difference was not statistically significant.

Intratumoral regional differences in DNA ploidy of gastrointestinal carcinomas.

Regional differences in DNA ploidy of gastrointestinal carcinomas were investigated by flow cytometric (FCM) analysis of multiple samples taken from different sites of each tumor of 42 patients. The overall incidence of DNA aneuploidy in the 42 cases examined was 80%, and the DNA index (DI) ranged from 0.825 to 2.170. However, intratumoral differences in DNA ploidy were observed in six of 15 cases (40.0%) of gastric carcinoma, whereas intratumoral differences in DNA ploidy were observed in only two of 27 cases (7.4%) of colorectal carcinoma. Therefore, to accurately determine DNA ploidy, one must analyze multiple samples from the tumor specimen (especially in gastric carcinomas) because cases exist where subpopulations with DNA aneuploidy are detected only at limited sites within the same tumor. The exact mechanisms responsible for the regional differences in DNA ploidy are not yet clear, although some hypotheses have been presented.

DNA aneuploidy in solitary colonic adenomas and the future risk of colorectal cancer.

Flow cytometric DNA analysis was performed on the colonic adenomas of 24 patients who had resection of an invasive colorectal carcinoma subsequent to polypectomy. The incidence of DNA aneuploidy among these adenomas was only 13 percent. Thus, simple recognition of colonic adenomas as being DNA diploid or DNA aneuploid is unlikely to be helpful for identifying patients with adenoma who are at high risk for developing a future colorectal carcinoma.

Differential pattern of DNA-Aneuploidy in human malignancies

The differential pattern of DNA-aneuploidy, detected by flow cytometry (FCM) regarding its frequency, grade and multiclonality, was investigated and correlated to tumor type, malignancy grade, tumor stage and prognosis in a multi-institutional study at the University of Münster. High resolution measurements using admixed normal blood reference cells were undertaken in 2413 cases of 13 different malignant diseases and in 776 benign lesions or samples. The incidence of DNA-aneuploidy was highest in melanomas, carcinomas, testicular tumors, sarcomas (75%-95%) and myelomas (65%). Acute leukemias showed an intermediate DNA-aneuploidy rate of 40% with special subgroups represented by common ALL (44%), p less than 0.05) and myelomonocytic/monocytic AML (47%, p less than 0.01). The lowest DNA-aneuploidy-rate was found in basal cell skin carcinomas (19%) and congenital melanocytic nevi (9%). No case of DNA-aneuploidy was observed in the 776 benign lesions or samples.--DNA-indices giving the grade of DNA-aneuploidy with 1.0 for normal diploid G1/0 cells were found distributed predominantly between 1.0 and 2.0 in the solid tumors, except testicular tumors, clustering around a triploid maximum at 1.5. DNA-indices of myelomas and acute leukemias generally ranged below 1.25 with lower DNA-aneuploidy grades in AML than in ALL (p less than 0.01).--In melanomas the aneuploidy rate was higher (86%) in metastases than in the primary tumors (54%, p = 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)