No abstract

Summary One hundred and fifty-seven patients with usual colorectal cancer were analysed prospectively for DNA-ploidy, DNA-index and S-phase fraction (SPF) using flow cytometry. An abnormal DNA-stemline was observed in 68% of tumours. The patients have been followed for a median of 36 months. In univariate analysis, tumour stage was the most significant prognostic factor. After excluding patients with stage D disease, DNA-aneuploidy was significantly associated with a shorter survival and a shorter disease free survival. SPF, however, did not correlate with prognosis. In multiple samples from the same tumour there was on average a 29% difference between the highest and the lowest SPF indicating considerable heterogeneity in proliferative activity within the tumours. In diploid tumours the variation was even higher. (Shapiro, 1989). This information has proved useful for supplementing clinical and pathological classification of many malignant diseases (Seckinger et al., 1989). An accumulating body of data supports the view that flow cytometric detection of an abnormal DNA content in cells and the degree of DNA abnormality and/or proliferative activity in most malignancies may reflect the degree of malignancy and clinical behaviour (Seckinger et al., 1989). In reports on DNA-abnormalities in colorectal carcinomas, aneuploidy correlates with lower survival rates than diploidy (Wolley et al., 1982;Armitage et al., 1985;Kokal et al., 1986;Goh et al., 1987; Scott et al., 1987a,b;Quirke et al., 1987;Jones et al., 1988), although the results are somewhat controversial (Melamed et al., 1986;Schutte et al., 1987;Wiggers et al., 1988). Also, a high S-phase fraction (SPF) has shown an association with poorer survival (Bauer et al., 1987).In (Davis & Newland, 1983). The tumours were classified as right-sided (e.g. those proximal to splenic flexure) left-sided and rectal.Follow-up Follow-up visits took place once every 6 months during the first 2 years, and then yearly thereafter up to 5 years. Routinely the visits included a physical examination, sigmoidoscopy and blood tests including serum carcinoembryonic antigen. Recurrence was dated from the first evidence of relapse, based on physical, histological or imaging data. The time of death and cause of death was recovered from hospital records and from the Central Statistical Office of Finland. The patients have been followed for a median of 36 months (range 13-67 months). Thirty-six of the 125 patients with stage A-C disease developed a locoregional and/or distant recurrence during the follow-up. Twenty-two of them died before the closing date. Twenty-six of 32 patients with stage D disease died before the closing date. In addition, 14 patients died without reported recurrence and were treated as censored observations in the analysis.Flow cytometric analysis Biopsies from each tumour for flow cytometric DNA-analysis were immediately frozen in liquid nitrogen and stored thereafter at -80°C until analysis. At the time of analysis, the tumour samples were rapidly thawed in a 3...

Fifty-nine colorectal carcinomas of patients with verified cancer family syndrome (CFS) were analyzed for DNA ploidy using flow cytometry. Sixty-eight percent of the tumors were diploid, and 32% were aneuploid. The aneuploid tumors had a median DNA index of 1.24 (range, 1.12-1.97). In 90% of all tumors the DNA index was less than 1.27. This predominance of diploid/near-diploid tumors was seen both in primary and in metachronous carcinomas. In 21 cases a cell cycle analysis was possible. Tumors with the S-phase fraction (SPF) 2 9.8% had a worse prognosis than tumors with the SPF of <9.8%. These findings suggest that the predominance of diploid/near diploid DNA values is one of the characteristics of colorectal carcinomas in CFS. This might signify the existence of two or more pathogenetically different subgroups of colorectal carcinoma and explain the proposed better prognosis of colorectal carcinoma in CFS compared with other colorectal carcinomas. Cancer 65:1825-1829,1990.ANCER FAMILY SYNDROME (CFS) is the most com-C mon verifiable genetic risk factor for colorectal car- been linked to poorer ~urvival."~'~ However, to our knowledge there have been no published reports separating DNA ploidy in colorectal carcinoma in CFS. The histologic characteristics of colonic neoplasms in CFS were recently reported in a series of 22 Finnish CFS kindreds.16 We undertook to determine if the DNA ploidy of the SPF of these tumors could explain the different clinical characteristics of colorectal carcinomas in CFS compared with the usual large bowel carcinomas. Patients and Methods Patients and TumorsSeventy-four patients (48 men and 26 women) with 100 colorectal carcinomas (diagnosed between 1952 and 1987) and verified CFS were studied retrospectively. The definition of CFS was the occurrence of at least three colorectal carcinoma cases in first-degree relatives. The pedigrees and details of clinical, genetic, and histologic features of 53 patients with 73 carcinomas were published previously. 16-18 Our series includes 2 1 new patients with verified CFS. Paraffin blocks were available from 81 tumors. Fifty-nine of them were evaluable for DNA ploidy. Twenty-two tumors were excluded from the flow cytometric DNA analysis, 12 of them because of an unsatis-1825

The present study was carried out in order to examine the radiosensitivity of malignant pleural mesothelioma cell lines. Cell kinetics, radiation-induced delay of the cell cycle and DNA ploidy of the cell lines were also determined. For comparison an HeLa and a human foetal fibroblast cell line were simultaneously explored. Six previously cytogenetically and histologically characterized mesothelioma tumor cell lines were applied. A rapid tiazolyl blue microtiter (MTT) assay was used to analyze radiosensitivity and cell kinetics and DNA ploidy of the cultured cells were determined by flow cytometry. The survival fraction after a dose of 2 Gy (SF2), parameters alpha and beta of the linear quadratic model (LQ-model) and mean inactivation dose (D(MID)) were also estimated. The DNA index of four cell lines equaled 1.0 and two cell lines equaled 1.5 and 1.6. Different mesothelioma cell lines showed a great variation in radiosensitivity. Mean survival fraction after a radiation dose of 2 Gy (SF2) was 0.60 and ranged from 0.36 to 0.81 and mean alpha value was 0.26 (range 0.48 - 0.083). The SF2 of the most sensitive diploid mesothelioma cell line was 0.36: less than that of the foetal fibroblast cell line (0.49). The survival fractions (0.81 and 0.74) of the two most resistant cell lines, which also were aneuploid, were equal to that of the HeLa cell line (0.78). The alpha/beta ratios of the most sensitive cell lines were almost an order of magnitude greater than those of the two most resistant cell lines. Radiation-induced delay of the most resistant aneuploid cell line was similar to that of HeLa cells but in the most sensitive (diploid cells) there was practically no entry into the G1 phase following the 2 Gy radiation dose during 36 h.

(Clark et al., 1969) and thickness of tumour (Breslow, 1970) are known to predict the clinical behaviour of primary melanoma.