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Background: The authors determined the flow cytometric (FCM) DNA characteristics of 53 benign tumors and 132 malignant (71 primary, 61 recurrent) soft tissue sarcomas to investigate their heterogeneity and to evaluate the prognostic values of DNA ploidy status and S-phase fraction (SPF)., Methods: One to 11 frozen samples were collected from 185 tumors at 10 participating centers of the French Federation of Cancer Centers (FNCLCC). All FCM analyses were performed in the same laboratory. Histologic diagnoses were collegially reviewed, and grade was assessed by the pathologists of the FNCLCC Sarcoma Group. Relationships between FCM, clinical, and pathologic data were investigated using univariate and multivariate analyses for risks of mortality and metastasis., Results: All except 2 of the 53 benign lesions were DNA diploid. One schwannoma and one desmoid tumor exhibited small abnormal DNA peaks. SPF was low in all benign lesions. One-third of the sarcomas were DNA diploid, whereas two-thirds were DNA aneuploid. Relationships were found between aneuploidy and mitotic count, grade, and histologic subtype of malignant fibrous histiocytoma. DNA ploidy status did not influence the clinical outcome. Multiple sampling performed in 32 sarcomas showed both diploid and aneuploid samples in 6 tumors. Heterogeneity was related to tumor size. SPF evaluated in 85 sarcomas was related to DNA ploidy status, mitotic count, and grade. SPF > or = 4% was significantly associated with a low overall survival rate. In a multivariate analysis performed for the whole group of 132 patients, the single factors with independent prognostic value for patient mortality were disease free status after the treatment course (P < 0.0001) and SPF (P = 0.03). In the subgroup of patients who were initially free of metastases and free of tumor after the treatment course, SPF remained the only factor that significantly influenced the overall survival rate (P = 0.021)., Conclusions: Despite its high failure rate, SPF is an independent factor in the prognosis of soft tissue sarcoma and should be considered when patients with a high mortality risk need to be selected for adjuvant chemotherapy.

In 132 soft-tissue sarcomas and 52 benign soft-tissue tumors, cellular proliferation was examined by immunohistochemistry using monoclonal antibodies Ki-S11 (Ki-67 antigen) and Ki-S1 (topoisomerase II alpha) and by flow cytometric analysis of the S-phase fraction (SPF). Malignant tumors were graded histologically according to the Fédération Nationale des Centres de Lutte Contre le Cancer (FNCLCC) system. Patient age, sex, tumor location, histological type, and DNA ploidy were considered as additional prognostic variables. Consistent immunoreactivity was seen in approximately 95% of the cases, and determination of SPF was possible in approximately 60% Ki-S11 and Ki-S1 immunolabeling indices correlated in a linear manner. All proliferation parameters yielded significant differences between benign and malignant tumors. Ki-S11 and Ki-S1 immunoreactive scores also co-varied significantly with SPF, mitotic count, and histopathological grade. In univariate analysis, immunohistochemical proliferation indices, histopathological grade, mitotic count, and SPF were predictive of overall survival and the development of metastases. In multivariate analysis, immunolabeling scores of proliferation markers, grade, and SPF emerged as independent predictors of global survival and systemic progression. We conclude that the immunohistochemical assessment of proliferation, being more readily performable and more easily assessable than the equally relevant S phase fraction, may add appreciable information to the current prognostic models for soft-tissue sarcoma.

To assess the prognostic value of flow cytometric analysis, 61 cases of operated squamous cell lung carcinoma were studied in terms of desoxyribonucleic acid (DNA) nuclear content and percentage of cells in DNA synthesis phase (% S). These parameters were determined on a fresh surgical sample. The S % was obtained in 25 cases. DNA index (DI) and % S were compared with survival and usual clinicopathologic characteristics for the prediction of survival. DNA content classified as DNA-diploid and DNA-aneuploid is not a prognostic factor for survival. DI higher than 2 seems to be predictive but needs confirmation. The % S and multiploidy are not predictive factors for survival.

When N-myc copy number was assessed by molecular biology, it has been proven that an amplification of the oncogene was a bad prognosis in childhood neuroblastoma, and the same goes for DNA-diploidy. This study concerns the development of a biparametric flow cytometric analysis of 2 neuroblastoma cell lines (SK-N-SH and IGR-N-91), which exhibited respectively 1 and 60 copies of the N-myc oncogene. An indirect immunofluorescence technique allowed N-myc oncoprotein staining and an isotypic control was used to assess the threshold of specific fluorescence. Simultaneously, a double staining with propidium iodide gave the nuclear DNA content. For both types of cells, the level of N-myc expression was calculated as a fluorescence index (IF). IF for IGR-N-91 appeared 2.5 times higher than those of SK-N-SH. This fluorescence index increases significantly during the exponential growth of N-myc amplified cells, whereas it does not vary for SK-N-SH. During IGR-N-91 tumoral evolution, the cell line which derived from murine heart metastasis was the only one to show an increased IF. When applied to 10 neuroblastoma cell suspensions, this double staining showed an high IF for only 1 N-myc amplified case. A poor cell yield after tumoral dissociation and too much debris did not allowed the calculation of IF for half of them, which hampered a routine development of this technique.

The authors determined the desoxyribonucleic acid (DNA) content of 45 small-cell lung cancers, by flow-cytometry (FC), from bronchial brushings to asses the prognostic meaning of the DNA index, the percentage of cells in DNA synthesis phase (% S) and the ploidy. Moreover, the value of this tumorous cells brushing was assessed for Flow-cytometric analysis. Bronchial brushing offers some advantages opposite to biopsies, but the low quantity of collected cells reduces the number of tumors analysable by FC. The output is low (39%) for determination of DNA index, and moreover low (22%) for determination of % S. In regard to the flow-cytometry analysis, the DNA content does not show any significant difference of survival where as the % S is related to survival, but it seems to be paradoxical in this study. Indeed, patients with small-cell lung cancer associated to high cells' percentage in DNA synthesis phase have the longest survival.

The flow-cytometry (FCM) is a method of quickly determining the desoxyribonucleic acid (DNA) content of tumoral cells. This provides us with two results: the DNA-ploidy which characterizes quantitatively the DNA, and the percentage of cells in DNA synthesis phase (%S). The contribution of FCM in evaluation of the prognosis of non-small cell lung cancer remains disputable. The absence of standardization between laboratories makes it difficult to compare the different studies to define the place of FCM analysis. However, for epidermoid carcinomas, the most actively studied, a DNA diploidy and a weak %S have a favourable impact on survival. No conclusion can be drawn from the studies on adenocarcinomas. Large cell carcinomas have never been studied in an isolated manner, due to the lack of numbers.

Flow cytometric DNA analysis was performed in 35 patients with squamous cell carcinoma of the esophagus. The aim of this study was a) to establish a tumoral DNA pattern, b) to determine an objective parameter correlating with tumoral response to chemotherapy (5 FU-cisplatin). DNA analysis was performed in perendoscopic ranged tumoral biopsy specimens to determine the DNA ploidy (DNA index) and S phase fraction. Tumor diameter and length were evaluated by computed tomography (CT) before and after chemotherapy in 24 patients. The relative variation of this product determined a CT index. Before chemotherapy, 82 percent (76/93) of the specimens were available for assessment; 72 percent (26/36) of the tumors were aneuploid. In these tumors, the DNA index ranged from 1.23 to 2.80. Five tumors had two distinct and simultaneous aneuploid populations. CT index values were not significantly different according to the ploidy (diploid, aneuploid), the S phase fraction (low, high), the DNA content modification after chemotherapy (absent, present). In this study, DNA analysis did not allow to select patients with higher response to chemotherapy. The inter- and intratumoral phenotypic heterogeneity may be one of the responsible factors.

Flow cytometry DNA analysis of fragments of fresh or paraffin-embedded tumor was performed in order to investigate its prognostic value in renal cell carcinoma. As aneuploidy is a frequent cytogenetic finding of these cancers, it is not surprising that the literature reports a high incidence of anomalies of DNA content, affecting 2/3 of cases studied. DNA aneuploidy is more frequent in advanced stages, high grades and spindle-cell carcinomas. Its prognostic value in terms of the survival of nephrectomized patients has been demonstrated by unifactorial studies. Several studies using Cox's model have demonstrated its independence of stage and grade, but they are too limited for DNA ploidy to be considered to be an element of the therapeutic decision. In contrast with other sites, the prognostic influence of the proliferation evaluated by S% does not appear to be as important as DNA ploidy. Prospective studies comparing these results should take into account the tumor heterogeneity observed in this type of cancer by analysing, for example, several fragments of the same tumor.

Cancerembryonic antigen (CEA) and beta2-microglobulin (beta2m) have been measured in cancer patients and patients with benign diseases. Of 168 patients with intestinal cancer, almost 90% had increasing concentrations of either CEA or beta2m or both. In 29 patients at different stages of pancreatic cancer there was a high incidence of increased values in the more severe cases. In 60 patients with histologically classified colorectal cancer the TNomegaMomega group of 19 patients had 47% and 42% of elevated beta2m and CEA respectively. A significant correlation of beta2m or CEA to extension of disease was noted. In benign intestinal disease like cirrhosis and pancreatitis both beta2m and CEA is commonly elevated. Of 26 breast cancer patients, seven had elevated CEA and five had elevated beta2m values before treatment. In the patients with extraganglionary metastasis almost 90% had high beta2m or CEA or both. Of 40 patients with uterine cancer, 26 were found to have increased values of beta2m or CEA or both. Finally, 140 colorectal cancer patients, 62 patients with breast cancer and 10 patients with uterine cancer have been followed longitudinally.

The results are reported of cytofluorometric DNA analyses of colorectal tumors using cells obtained by mechanical disruption and fine needle aspiration. The latter method does not lower the level of debris or cell aggregates. We found a significantly higher proportion of aneuploid cells by needle aspiration. There was a good correlation between the data obtained by both methods, but for large heterogeneous tumors multiple site aspirations are needed to obtain representative specimens. Fine needle aspiration was found particularly useful for small tumors and polyps.

We report a flow cytometric study on ploidy in 117 colorectal cancers. An aneuploid cell population was found in more than 70% of adenocarcinomas. Ploidy was found to be stage-related; aneuploid tumors with DNA index greater than or equal to 1.4 were found mainly in Dukes' C and Dukes' D stages (P less than 0.02). Tumors of the caecum and of the ascending colon were more often found to be diploid than those of the other sites. There was a progressive increase in the proportion of cells in S-phase depending on whether they were from normal tissue, inflammatory mucosa or adenocarcinomas. The proportion of cells in S-phase was significantly larger in aneuploid tumors (P less than 0.001). The data presented above suggest that aneuploidy and the proportion of non-resting cells could be important prognostic factors for colorectal cancers. The latter are independent of the stage of the disease and histological differentiation.

The efficiency of flow cytometric (FCM) DNA analysis in the diagnosis of lung carcinoma was compared with that of conventional cytologic techniques on bronchial brushing and fine needle aspiration samples from 461 patients. The main advantage of FCM was the rapid delivery of results. Unfortunately, this was offset by a poor sensitivity in the detection of bronchial tumors. Nevertheless, DNA analysis may still prove useful in determining the prognosis and in evaluating the effects of chemotherapy on known tumor stem lines.