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The production of metastases depends on changes in a large number of genes. It is also connected with the interaction of tumor cells with the environment. It has been reported that primary tumor clone domination is also an important factor in metastasizing, and in many neoplasms dominating clones are the metastatic forerunners. Up to now it is unknown whether domination of a given clone in a primary renal cell carcinoma is a crucial factor in forming metastases or rather presence or absence of specific genes imposes the major advantage in the metastatic process. To study the presence or absence of the duplication and mitotic nondisjunction event as one of the phenomenon in the creation of metastases, as well as possible derivation of metastatic cells from the minor subclone of primary tumor, we examined three metastatic renal clear-cell carcinomas in which by comparative genomic hybridization we detected the loss of 3p in the primary tumor and two copies of 3p in the corresponding metastasis. Loss of heterozygosity analyses using markers for 3p25 (D3S1038), 3p21.1 (D3S1295), and 3p14.2 (D3S1481) proved heterozygosity of at least two 3p loci in all metastatic tumors, which indicates the absence of mitotic nondisjunction event as a cause of occurrence of two copies of 3p in metastases. Our results suggest that in some of the clear-cell renal carcinomas metastatic cells may derive from minor subclones of primary tumors.

Archival material from primary and metastatic renal clear cell carcinomas of 25 patients was studied by comparative genomic hybridization. Copy number changes of entire chromosomes or chromosomal subregions were detected in 22 primary and 21 metastatic tumors. Copy number changes affected the following chromosomes in at least 20% of the 25 primary tumors (minimal common region given in parentheses): gains were noted for chromosomes 1 (1q21-->q23), 5 (5q31-->q34), 7 (7p), 8 (8q), 16 (16p), 17 (17q12-->qter), 19, and 22 (22q12-->qter); losses were revealed for chromosomes 3 (3p21-->pter), 8 (8p23-->pter), 14(14q21-->qter), and Y. The same chromosomal regions that were involved in primary renal clear cell carcinomas were also found in the respective metastatic tumors but with strikingly different frequencies for a few regions. Metastatic tumors showed a significantly higher frequency of complete or partial gains of the long arm of chromosome 1, in particular at 1q21-->q23 than primary tumors (16 cases versus 6 cases; P < 0.005). These data suggest a correlation of metastatic events in renal clear cell carcinomas with an increase in the copy number of genes located at 1q, in particular at 1q21-->q23. In contrast, the entire or partial loss of the short arm of chromosome 3 was significantly less frequent in metastatic tumors (8 cases versus 15 cases; P < 0.025). The validity of 1q and 3p copy number changes detected by comparative genomic hybridization was confirmed by interphase cytogenetics with region-specific yeast artificial chromosomes to paraffin-embedded tumor tissue sections.

Background: The influence of DNA content on prognosis in stomach cancer has been investigated rarely, and the results are controversial., Method: The prognostic relevance of the DNA content and histomorphologic parameters was evaluated in 41 patients after resection of pancreatic cancer., Results: In the univariate analysis, the DNA content, tumor size, lymph node status, tumor stage, nuclear grade, and type of resection had a statistically significant influence on the prognosis. No association was found between the DNA content and the histomorphologic features. Apart from the operative procedure, the DNA content was the strongest indicator of prognosis in the multivariate analysis., Conclusions: Further investigations are necessary to find out if DNA analysis can be performed preoperatively on material obtained by fine-needle aspiration.

In 133 cases of patients with renal cell carcinoma which infiltrated locally into the renal fatty tissue (stage pT3a, TNM classification of 1987), the prognostic potential of the following parameters was investigated: symptoms, patient's age at the time of operation, tumor size and localization, grade of malignancy, cell type, growth pattern, and prognostic score according to Störkel et al. [Eur Urol 1990;18(suppl 2):36]. Additionally, automated image analysis DNA cytometry was performed on 110/133 carcinomas. After an average observation period of 3.6 years (maximum 10.1 years), 59 (44.4%) of the patients died of their tumors. The cause-specific 5- and 10-year survival rates were 51.3 and 29.1%, respectively. Statistically significant differences (p < 0.05) within the individual parameters were found only for the grade of malignancy and the corresponding prognostic score. Using DNA cytometry, 13 types of renal cell carcinoma were differentiated; 90% of the tumors contained aneuploid cells. Significant differences between these relative to prognosis did not exist. In the case of locally advanced renal cell carcinoma, the DNA histogram does not seem to be superior to conventional prognostic criteria.

The DNA histograms of 57 conservatively resected renal tumors were studied using automated image analysis DNA cytometry (Leytas II). Forty-nine of the analyzed tumors were renal cell carcinomas, six were oncocytomas, one was an angiomyolipoma, and one was a renal cell adenoma. On the basis of their DNA histograms, diploid, tetraploid, and aneuploid tumors could be distinguished. Aneuploid tumors could be subtyped further according to the DNA content of the stem cell line as hyperdiploid, hypertriploid, or hypertetraploid. Eight of the tumors were characterized by a combination of diploid and hypertriploid stem cell lines. During a mean follow-up of 5 years, only the two patients with a pure hypertriploid tumor died of distant metastases. These results indicate that automated DNA image analysis cytometry is able to differentiate among several types of renal tumors with obviously different prognoses.

In this study we analysed DNA-ploidy as a potential prognostic parameter in ductal pancreatic carcinoma. Paraffin embedded histological material, obtained by resection from 34 patients with a ductal pancreatic carcinoma, was selected for analysis. Tumor areas within the paraffin embedded material were identified by HE-stained reference sections. One 50 microns section was dewaxed, rehydrated and mechanically and enzymatically prepared to form a suspension of 10000 cells/ml. One milliliter of the suspension, which contained bare nuclei with small rests of cytoplasma, was centrifuged on glass slides. The fixed nuclei were air-dried and stained by Feulgen SITS technique, which allows for the quantitative measurement of DNA. The DNA analysis was carried out with a computer-controlled single-cell cytophotometry. In contrast to using flow cytometry, only the tumor cells were measured by image-cytometry. Overlapping nuclei, dirt and other artifacts as well as inflammatory cells were efficiently eliminated. With DNA image-cytometry, we could differentiate between hypotriploid, triploid, hypertriploid and tetraploid tumors. The DNA-content provided the strongest influence on prognosis in the multivariate analysis.

Esophagus cancer is a heterogeneous disease with considerable differences in malignant behaviour. Some relevant factors for prognosis are known. In this study we analyzed DNA-ploidy as a potential prognostic parameter in esophagus carcinoma. Paraffin embedded histological material from 50 patients with an esophagus cancer, obtained by resection, were selected for analysis. Tumor areas within the paraffin material were identified by HE-stained reference sections. One 50 microns section was dewaxed, rehydrated and mechanically and enzymatically treated to a suspension of 10,000 cells/ml. 1 ml of the suspension, containing bare nuclei with small rests of cytoplasma was centrifuged on glass slides. The fixed nuclei were air-dried and stained by Feulgen-SITS technique, which allows quantitative measurement of DNA. The DNA analysis was carried out with a computer-controlled single cell cytophotometry (Leytas 2, Leitz, Wetzlar). In contrast to the flow cytometry with image cytometry only tumors cells were measured. Overlapping nuclei, dirt and other artefacts as well as inflammatory cells were efficiently eliminated. With the DNA image cytometry we could differentiate between diploid and hypotriploid, hypertriploid aneuploid tumors. Best prognosis had diploid and hypotriploid tumors, the worst hypertriploid carcinomas. In the multivariate analysis the DNA-content of the tumor cells in esophagus cancer was the only prognostic parameter. DNA-content of tumor cells may become considerably clinical relevant in esophagus cancer for the decision to perform a resection or palliative treatment. In patients with hypertriploid tumors an adjuvant oncological therapy may increase the prognosis.

In 45 patients who underwent an esophagus resection due to a squamous cell carcinoma, in addition to the TNM classification and usual morphologic criteria, the paraffin-embedded material underwent deparaffinization, was rehydrated, and was mechanically and enzymatically processed into a single-cell solution. For evaluating the DNA histogram this was analyzed with the help of automatic single-cell cytophotometric study. The method, contrary to that of flow cytometric study, allows for the selective analysis of tumor cells due to the electronically, previously given selection criteria, whereas artifacts, stroma, and infection cells remain excluded from analysis. The multivariate analysis shows that the prognosis is only correlated with DNA content of the tumor cells. Patients with diploid tumors had a median survival time of 32 months as compared with the 22 months of patients with hypotriploid tumors, and 6.5 months with hypertriploid tumors. DNA cytometric analysis should be included when diagnosing patients with esophagus carcinoma.

The surgical technique, course and complications were analysed in 123 patients in whom a kidney tumor had been enucleated. Fifty-seven enucleated tumors were available for image analysis DNA-cytometry. In 49 patients there was an imperative indication for an organ-preserving operation, because nephrectomy would have made dialysis obligatory. In 74 patients with a healthy contralateral kidney the tumor was enucleated by choice. Thirty-five of 49 patients with an imperative indication are without sign of tumor progression after a mean follow-up of 4.5 years. In two patients there was a recurrence after 4 and 5 years, respectively, requiring a second organ-preserving operation. In one further patient there is a suspicion of multiple small tumor lesions 2 years after the first operation. Known metastases were present in 3 of 6 patients who died of their tumors. Sixty-eight of 74 patients operated on electively are without signs of tumor progression after a mean follow-up period of 3.3 years. One patient died from tumor metastases. Two patients had tumor recurrence, requiring nephrectomy and enucleation, respectively.