DNA ploidy pattern of gastric carcinomas and its relations with histological type and invasion pattern

The DNA ploidy pattern was determined by cytofluorometry in 90 cases of surgically resected gastric carcinomas, which were histologically divided into 40 cases of differentiated type and 50 of undifferentiated type. The gastric cancers were classified into two basic ploidy patterns, diploid and aneuploid. Relations among ploidy patterns, histological types and invasion patterns were studied. Diploid patterns were found in 23 cases (58%) of differentiated type and in 37 (74%) of undifferentiated type. In undifferentiated types, aneuploid patterns appeared in 2 (10%) of 19 early cancers, whose rate was significantly lower than that of advanced cancers. 11 (38%) of 29. In differentiated types, however, aneuploids were seen in 10 (48%) of 21 early cancers, not lower than 7 (37%) of 19 advanced ones. Furthermore, in undifferentiated types, aneuploids marked 11 (52%) out of 21 highly invasive carcinomas, while no aneuploids appeared in slightly invasive ones. In differentiated types, aneuploids were found in 3 (38%) out of 8 intramucosal carcinomas and 9 (53%) out of 17 slightly invasive ones. These results suggested that in undifferentiated gastric carcinomas, aneuploid is closely correlated with tumor progression and invasion degree, whereas in differentiated carcinomas such a correlation is not clearly seen.     

Cell cycle parameters and DNA ploidy in colorectal carcinomas

Seven patients with colorectal adenocarcinoma and one with squamous cell carcinoma of anorectal region were infused preoperatively with iododeoxyuridine (IUdR) and bromodeoxyuridine (BrdU) in sequence (100 mg/m2 x 1 hr each with 1-hr interval in between) to label S-phase cells. The tumor biopsy specimens were embedded in glycol-methacrylate and 2-microns thick sections were treated with two monoclonal antibodies which permitted the identification of cells which incorporated IUdR only, BrdU only, both IUdR and BrdU, or neither IUdR or BrdU. The labeling index of tumors varied from 17.3 to 35.6% (mean = 25.78 +/- 6.162), duration of S-phase ranged from 14.0 to 23.9 hr (mean = 18.73 +/- 3.712), and total cell cycle time ranged from 39.4 to 123.4 hr (mean = 76.78 +/- 24.165). The architecture of the tumor was well preserved and a variable number of DNA synthesizing mononuclear cells were identified within and around the tumor. Image analysis of Feulgen-stained smears of the tumors was done to measure the DNA content of seven tumor samples. Each tumor was found to be hyperdiploid with multiple modal values. The studies described here demonstrate the feasibility of performing cell cycle kinetic measurements on gastrointestinal tumors which have been labeled in vivo. The ability to perform these measurements on tumor biopsies allows the avoidance of artifacts introduced when solid tumors are disaggregated in vitro for study.     

DNA ploidy pattern and tumour spread in gastric cancer

The DNA ploidy pattern of gastric cancer was studied in 58 patients to investigate the heterogeneity between primary tumour and metastases. In both primary tumours and lymph node metastases, diploid patterns accounted for 33 per cent, whereas all liver metastases were aneuploid. The percentage of polyploid cells was higher in the liver metastases than in primary tumours and lymph node metastases. When the heterogeneity of DNA ploidy pattern between primary tumour and metastasis was evaluated, diploid tumours had a significantly lower rate of lymph node metastasis heterogeneity than aneuploid tumours. When the DNA ploidy pattern and survival were evaluated, the patients who had a diploid pattern in both primary tumour and metastasis had a significantly higher survival rate than the patients who had an aneuploid pattern in the primary tumour and metastasis (57 per cent versus 26 per cent at 5 years). These data suggest that cell heterogeneity is a common phenomenon in gastric cancer, and this may be important in the evolution of the disease. Furthermore, the role of the DNA ploidy pattern as a prognostic factor is emphasized.     

DNA ploidy in oral cavity carcinomas,with special reference to prognosis

Single-cell DNA cytofluorometry was performed on paraffin-em bedded tissue of 140 patients with squamous cell carcinomas of the oral cavity. Half of the tumors (71 of 140) were DNA nondip loid. Well-differentiated carcinomas were more often DNA dip loid than moderately well-differentiated ones (P<0.001; chi square). The aneuploid tumors responded better to preoperative radiotherapy than did the DNA diploid (P<0.001) and polyploid tumors (P<0.05; chi-square). Using the multivariate Cox's re gression analysis multiploid type tumor, age of the patients and presence of lymph node metastases were the only significant factors influencing survival. DNA diploid tumors in stages I and II had a better prognosis than DNA nondiploid (P<0.01; Kaplan-Meier). The reverse was true for stages III and IV, where DNA nondiploid tumors had a better prognosis (P<0.05; Kaplan-Meier). Tumor stages (P<0.001; Kaplan-Meier) and especially lymph node metastases (P<0.0005; Kaplan-Meier) were major prognostic factors. Tumor DNA ploidy may be a complement to clinical and morphologic parameters as a prognostic predictor in squamous cell carcinoma of the oral cavity.     

DNA ploidy in submucosal cancer of the stomach and its relationship to lymph node metastasis

The relationship between DNA ploidy and lymph node metastasis was determined in 40 cases of gastric cancer confined to the submucosa (with lymph node metastasis 20 cases and without 20 cases). The DNA ploidy patterns were classified as follows: Type D, Type A1 and Type A2. Of the 20 cases with lymph node metastasis, 1 was Type D, 7 were Type A1 and 12 were Type A2. The likelihood of lymph node metastasis was 12.5% (1/8) for Type D, 43.8% (7/16) for Type A1 and 75.0% (12/16) for Type A2. It is concluded that although gastric cancer confined to the submucosa is classified as early one, analysis of DNA content places such tumors with lymph node metastasis into the advanced cancer category.     

Expression of human epidermal growth factor and DNA ploidy pattern in gastric carcinoma

Expression of human epidermal growth factor (EGF) was examined immunohistologically in 93 surgically resected gastric carcinomas, using Biotin-StreptAvidin method pretreated by protease, and its relation with cancer progression and DNA ploidy pattern was studied. DNA ploidy pattern was determined by cytofluorometric measurement. The gastric cancers were divided into two basic ploidy patterns; a diploid mode and an aneuploid mode. EGF expressions were found in 5 out of 20 cases (25%) in intramucosal carcinomas and, the more deeply the carcinomas invaded into the gastric wall, the greater the frequency of EGF expressions did not always become. In the carcinomas with the aneuploid pattern, EGF expression was found only in 2 out of 12 cases (17%) in early cancers and significantly increased in advanced cancers, 15 out of 22 cases (68%). On the other hand, in the carcinomas with the diploid pattern, EGF was expressed in 9 out of 31 cases (29%) in advanced cancers, not significantly different in early cancers, 11 out of 28 cases (39%). Consequently, in advanced cancers, EGF was found more frequently in the aneuploid tumors than in the diploid tumors. These results suggested that EGF expression in the gastric carcinomas is closely correlated with the progression to the advanced cancer with DNA aneuploidy.     

Malignancy of gastric cancer according to DNA ploidy pattern and tumor markers

Microfluorometric analysis of DNA content was performed on samples from 172 gastrectomy specimens of cancer. Histograms of DNA content were classified into three ploidy patterns; type I, II and III. Also, immunocytochemical examination for tumor markers (CEA, AFP and hCG) was performed. Type I was predominant in intramucosal carcinoma. Type III was predominant in differentiated carcinomas, in elevated type of early carcinomas and in gross type 1 and 2 of advanced carcinomas. Prognostic serosal factor exerted a greater influence on the prognosis than ploidy pattern. But in the patient without prognostic serosal factor, type III had a poorer prognosis than the other types because of vessel invasion, lymph node metastasis and liver metastasis. AFP and hCG were highly positive in type III. Furthermore type III cancer producing more than two tumor markers had an extremely poor prognosis. Analysis of ploidy pattern and tumor markers may prove to be a highly useful adjunct in the evaluation of malignancy in gastric cancer.     

流式细胞术分析肝细胞癌DNA倍体异质性的研究

目的 研究肝细胞癌(肝癌)是否存在DNA倍体的异质性。方法 采用流式细胞术检测了29例肝癌标车不同区域的DNA倍体情况。结果 16例为异倍体，13例为二倍体，没有发生二倍体和异倍体并存的肝癌，16例异倍体肝癌中，9例肿瘤内DNA指数相同．其他7例都存在异倍体亚克隆。结论 在肝癌的发生发展过程中二倍体肿瘤和异倍体肿瘤各自按固有的倍体模式增殖，二倍体肝癌一般不转化为异倍体肝癌，但异倍体肝癌可产生新的异倍体亚克隆。     

Flow cytometric DNA analysis of malignant potential in colorectal carcinoma--DNA ploidy pattern and liver metastasis

On the purpose of elucidating the malignant potential, flow cytometric DNA analyses were carried out using paraffin-embedded materials of 82 subserosal, serosal and adventitial invasive colorectal carcinoma. The ratio of DNA diploidy against DNA aneuploidy was one to two. DNA ploidy pattern was not correlated with histological grade but with histopathological factors such as lymphatic permeation, lymph node metastasis and venous invasion. At the primary operation, the rate of the liver metastasis in DNA aneuploid cancer was 16.4% but in DNA diploid cancer the liver metastasis was not observed at all. On the liver metastasis, the 5-year disease free survival rates were lower in patients with DNA aneuploidy (72.8%) than those with DNA diploidy (95.2%). These results indicate that DNA ploidy pattern is related to liver metastasis. Furthermore, the 5-year survival rates in patients with absolute curative resection were lower significantly in DNA aneuploidy (62.5%) than DNA diploidy (92.9%). In conclusion flow cytometric DNA analysis is useful for evaluating the biological malignant potential and predicting the liver metastasis in colorectal carcinoma.     

Correlation between DNA ploidy patterns and tumor progression, prognosis, and tumor infiltrating lymphocytes in human esophageal cancer

Cytophotometric DNA analyses were performed on 35 primary esophageal cancer. Histograms of DNA measurement were classified into three patterns (diploid pattern, aneuploid pattern and mosaic pattern) and were compared with histological findings, prognosis, and degree of lymphocytes infiltration around the tumor. Survival rate was worse in patients with mosaic pattern than the others, and 4-year survival rates of each patterns were 66.7% (diploid), 53.6% (aneuploid) and 25.4% (mosaic). Diploid cell line was observed frequently in the superficial cancers, and as the cancers infiltrated more deeply, mosaic cell line increased. Mosaic cell line appeared more frequently in well differentiated squamous cell carcinomas. In patients with mosaic pattern, there was high frequency of lymphnode metastasis and vascular invasion, as compared with diploid pattern. The rate of vascular invasion tended to increase in the following order; diploid, aneuploid and mosaic types. Furthermore in the diploid tumors, the degree of lymphocyte infiltration around tumor tended to increase. These findings suggest that the change of DNA content may occur frequently during tumor progression and may be affected by tumor infiltrating lymphocytes. So the DNA ploid pattern will be also to be one of the conjecturable factors of the prognosis of esophageal cancer.     

DNA ploidy in papillary tumours of the breast

The DNA content of benign and malignant papillary tumours of the breast as well as a group of papillary tumours where this distinction could not be made with certainty was measured by flow cytometry. All the benign papillomas were diploid; 5 of 19 carcinomas were aneuploid; and 1 of 15 tumours of undecided type was also aneuploid. Comparison of the DNA indices, proliferation indices and coefficients of variation were of no value in distinguishing between benign and malignant specimens.     

Flow cytometric analysis of DNA ploidy pattern in advanced gastric cancer and its relationship with prognosis

Cell nuclear DNA content was determined by flow cytometric analysis in 270 patients with advanced gastric cancer. Aneuploid DNA content was observed in 150 patients (55.6%). The DNA ploidy pattern was the third significantly prognostic factor behind peritoneal dissemination and liver metastasis in Cox regression multivariate analysis. About the relationship between DNA ploidy pattern and other prognostic factors, peritoneal dissemination and wall invasion ratio of aneuploid were significantly higher than those of diploid (p less than 0.01). Five-year survival rate of diploidy patients was significantly higher than that of aneuploidy patients. In stage I, five-year survival rate of patients with diploid tumor was 83.3% and that of patients with aneuploid tumor was 70.0%. In stage II, that of patients with diploid tumor was 81.3% and that of patients with aneuploid tumor was 66.7%. In stage III, that of patients with diploid tumor was 71.2% and that of patients with aneuploid tumor was 25.1%. In stage IV, that of patients with diploid tumor was 31.6% and that of patients with aneuploid tumor was 2.6%. Furthermore in the curative case, that of patients with diploid tumor was 77.2% and that of patients with aneuploid tumor was 48.2%. Aneuploid case has significantly worse prognosis in curative operation.     

The prognostic role of the DNA ploidy pattern in colorectal cancer analysis using paraffin-embedded tissue by an improved method

To assess the prognostic value of DNA ploidy in colorectal cancer, compared with the histopathological findings, paraffin-embedded surgical specimens from 330 patients who underwent resection for primary adenocarcinoma were studied using a new modified method of flow cytometry. Of these specimens, 141 were DNA diploid and 189, DNA aneuploid, among which there were 3 DNA hypodiploid lesions. Of the ten variables studied in curative resection, DNA ploidy ranked fourth in prognostic significance according to the linear trend by the ₂ test, after nodal status, grade of cellular differentiation, and degree of invasive growth, if the DNA ploidy pattern was classified into three categories. Conversely, DNA ploidy was the sixth most significant factor if DNA hypodiploidy was included in the DNA aneuploidy. The Cox multivariate analysis showed that DNA ploidy was one of the five significant factors independently determining prognosis; however, if adjustment for the modified Dukes' stage was made by the Mantel-Haenszel test, the survival difference between the diploid and aneuploid groups did not reach a statistically significant level. Thus, we conclude that from a practical point of view, DNA ploidy is not an essential factor which must be combined with histopathological variables for a better prediction of patient outcome.     

Therapeutic approach and associated problems of advanced liver tumors in children based on nuclear DNA ploidy pattern

We evaluated the therapeutic approach and its associated problems in 14 hepatoblastomas (HB) and 3 hepatocellular carcinomas (HCC) based on the ploidy patterns determined from paraffin-embedded tissues by flow cytometry. Aneuploid pattern was seen more frequently in the patients with poorly differentiated and immature types of HB, and HCC. It was also observed in patients with tumors showing microscopic vessel invasion, and in patients showing resistance to the therapeutic approach. Among 4 patients with combined epithelial components of HB, 2 patients' tumors revealed both the aneuploid and diploid patterns and their prognoses were poor. The prognoses of patients with tumors showing the aneuploid pattern were significantly poorer. From our study we concluded that careful attention should be pain to the heterogeneity of these tumors and that more aggressive treatment for tumors showing aneuploid pattern should be developed.     

Development and progression of undifferentiated carcinomas in the stomach]

Pathohistological studies of resected human stomachs and of experimental gastric cancers induced by ENNG have revealed that undifferentiated carcinomas arise at the neck region of glandular tubules both in the fundic and the pyloric mucosa, and tumor cells disclose the earliest invasion in the lamina propria by dripping from the glandular tubule. At earlier stages, the carcinoma cells tend to be confined to the middle level of the mucosa, and they extend to the horizontal direction of the mucosa. Most carcinomas at earlier stages comprise the diploid cell line. When tumors grow beyond a size of 2 cm in diameter in the mucosal layer, they begin to invade into the submucosal layer. As tumors grow, aneuploid and polyploid cancer cells arise in the diploid cell population. This is a kind of tumor progression. Aneuploid cancer cells disclose a more invasiveness, and they are ready to invade into the deep layer of the gastric wall. Scirrhous cancers are mostly composed of aneuploid cells, and it is suggested that small mucosal cancers which exclusively consist of aneuploid cells may become scirrhous cancers in a relatively short period.     

DNA ploidy in bronchopulmonary carcinoid tumours.

Fifty three bronchopulmonary carcinoid tumours were studied to assess the significance of DNA ploidy, determined by flow cytometry of paraffin embedded tissue. Twenty eight were typical carcinoid tumours and 25 well differentiated neuroendocrine carcinomas. Twenty seven were DNA diploid and 26 DNA aneuploid. DNA aneuploidy was significantly associated with histological features of increased malignant potential. Survival data were available for 43 patients. Of the 19 with DNA diploid tumours, 16 survived five years, compared with 14 of 24 with DNA aneuploid tumours--the difference being at the borderline of statistical significance. In a Cox multivariate regression analysis with other histological variables, DNA ploidy did not confer independent prognostic information. It is concluded that, although DNA aneuploidy as determined by flow cytometry is an indicator of increased malignant potential in bronchopulmonary carcinoid tumours, it does not provide clinically useful information additional to the results of routine histological examination.