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We determined DNA content, S-phase fraction, and estrogen (ER) und progesterone receptor (PR) levels in 36 stage I endometrial adenocarcinomas and in 22 hyperplastic lesions to obtain information on the genesis and progression of endometrial malignancy. DNA aneuploidy was detected in 12/36 (33%) carcinomas and in none of the hyperplastic lesions. DNA aneuploidy was significantly more common in poorly and moderately differentiated carcinomas than in the well-differentiated ones. Similarly, the highest number of cells in S-phase were found in poorly and moderately differentiated carcinomas, whereas well-differentiated carcinomas and all hyperplasias had an equally small S-phase fraction. Mean ER and PR levels were highest in hyperplastic lesions, especially those with atypical features, whereas carcinomas of all grades had significantly lower values. Thus, it is likely that the loss or decreased expression of steroid receptors is an early event during carcinogenesis in human endometrium, whereas an increase in the cell proliferation rate and the formation of DNA aneuploidy occur later during tumor progression.

We analysed ploidy and S-phase fraction (SPF) from 78 paraffin-embedded primary prostatic carcinomas by DNA flow cytometry. DNA aneuploidy and above median (4.2%) SPF were both associated with high tumour grade, large size of prostate and presence of distant metastases. Both aneuploidy and high SPF (greater than 4.2%) indicated short 10-year progression-free (P = 0.01 for ploidy and P = 0.0002 for SPF), overall (P = 0.004 and P less than 0.0001) as well as prostate cancer survival (P = 0.002 and P less than 0.0001). Ten-year overall survival rate was 45% in cases with SPF below 4.2% and 0% in those with higher values, whereas the corresponding prostate cancer-specific survival rates were 80% and 11%, respectively. None of the seven tumours with SPF above 12% showed an objective response to endocrine therapy, whereas 26/49 (52%) of those with lower SPF values responded (P = 0.01). DNA ploidy, tumour grade, T-stage or M-stage did not significantly correlate with endocrine responsiveness. SPF was also the best predictor of progression free survival in patients treated hormonally. In conclusion, detection of high SPF in prostate cancer may indicate lack of hormonal responsiveness and poor prognosis.

Consecutive sections from 33 paraffin-embedded human breast carcinomas without intratumor heterogeneity were sent for flow cytometric (FCM) DNA analysis in two experienced laboratories. FCM instruments, run conditions, and tumor disaggregation procedures were different in the two laboratories. In four cases (12%) the laboratories reported a different DNA ploidy and DNA index (DI). These variations were due to analytical reasons, differences in the detection rates of near-diploid and tetraploid DIs, not due to interpretation of data or the criteria used for aneuploidy. There was a significant correlation between S-phase fractions (SPF) obtained in the two laboratories (r = 0.90, p less than 0.0001) if only cases with concordant DI were included. Discordant DI usually led to very different SPF values.

An organized mammographic screening program covered 8,690 women from selected birth cohorts (aged 50-59) in the Tampere University Central Hospital district. Forty-four breast cancer cases were detected in the first round of mammographic screening, which is 3.7 times the expected annual number of new cases in this population. To evaluate the proliferative kinetics and biological properties of these cancers, DNA flow cytometric analysis was carried out in 37 of the screen detected cancers (SDCs) using 60 clinically detected stage I-II cancers and 30 screen-detected benign lesions as reference. DNA aneuploidy was observed in 17/37 (46%) of the SDCs as compared to 41/60 (68%) in the clinical controls (p less than 0.05), while all the benign lesions were DNA-diploid. The median S-phase fraction (SPF) in the SDCs was significantly (p less than 0.001) lower (3.5%) than in the clinical controls (9.6%). Differences in SPF persisted in subgroups defined by DNA ploidy and histological type. In stage-I SDCs the median SPF value (2.5%) approached that of benign tumors (1.9%). Our epidemiological and biological data indicate that the first round of mammography predominantly detects prevalent preclinical lesions, some of which are of very low malignant potential. At present such patients may often receive too extensive treatment. DNA flow cytometry could help in the identification of cases which could be treated, for example, by breast-conserving methods.

We determined nuclear DNA content from 308 archival paraffin-embedded malignant breast tumours and evaluated the survival of the patients by univariate and multivariate statistical analyses. The overall 8-year survival rate of stage I-III breast cancer patients was 74.3% in DNA-diploid and 51.2% in DNA-aneuploid tumours (P less than 0.0001). DNA ploidy had prognostic significance in both node-negative and node-positive breast cancer, primarily in cases with steroid receptor-positive tumours. In a Cox multivariate analysis DNA ploidy (P = 0.001), primary tumour size (P = 0.0007), nodal status (P = 0.04) and the content of progesterone receptors (P = 0.0008) emerged as significant independent prognostic factors, whereas oestrogen receptor status, age and menopausal status of the patients had no significant independent prognostic value. If the histological grade of ductal carcinomas was also included in the Cox model, both grade and DNA ploidy had independent prognostic effect. In conclusion, our results indicate that the analysis of DNA ploidy is a useful adjunct in the assessment of prognosis for breast cancer patients.

Eleven invasive and five non-invasive primary vaginal carcinomas were studied by DNA flow cytometry using archival paraffin-embedded tissue as starting material. Overall frequency of DNA aneuploidy in the invasive carcinomas was 8/11 (73%). DNA aneuploidy occurred in all four advanced stage (III-IV) and in 4/7 (57%) of the early stage (I-II) carcinomas. Among the squamous cell carcinomas aneuploid DNA content was also associated with non-keratinizing tumor type. Invasive vaginal carcinomas showed a high median S-phase fraction (SPF) (18.4%, range 6.9-31.8%). High SPF values were associated with advanced stage and non-keratinizing tumors. Corrected 5-year survival rate in invasive vaginal cancer was 44%, with no significant relation to DNA ploidy or SPF. In situ carcinomas were almost as often DNA-aneuploid (3/5, 60%) as the invasive carcinomas and had comparable median SPF value (13.4%, range 5.5-24.6). One in situ carcinoma with a high DNA-index and SPF relapsed, but overall 5-year survival rate was 100%. In conclusion, both invasive and in situ vaginal carcinomas frequently contain DNA-aneuploid stemlines and show a high SPF. Although DNA aneuploidy and high SPF correlate with advanced stage and non-keratinizing tumor type, they do not have much prognostic relevance in vaginal neoplasia.

We have previously defined three types of tumor DNA histograms, which are associated with favourable, intermediate and poor prognosis of patients with ovarian cancer. In the present study we evaluated the value of DNA histogram type combined with clinicopathological data in predicting long-term clinical outcome in stage I ovarian cancer. A stepwise discriminant analysis was done to find out the best combination of prognostic parameters in the distinction of two groups of stage I ovarian cancer patients; those with less than 5-year overall survival (22 cases) and those with longer than 5-year recurrence-free survival (47 cases). DNA histogram and histological type as well as FIGO stage in that order proved to be the most discriminating parameters and their combination allowed the correct prediction of clinical outcome in 78% of stage I ovarian cancer patients. In stage Ia the proportion of correctly classified cases based on DNA histogram and histological type was 82%. If DNA histogram was omitted from the discriminant analysis, the combination stage and histological type correctly classified a significantly lower percentage (68%) of patients. DNA flow cytometry thus improved the prognostic evaluation in stage I ovarian cancer, but even when combined with conventional clinicopathological factors failed to give correct prognostic assessment in about 20% of patients with stage I ovarian cancer.

To optimize the prognostic value of DNA flow cytometry in breast cancer the authors calculated several parameters from the DNA histogram, including the DNA index, the size and number of aneuploid peaks as well as S-phase and G2/M-phase cell cycle fractions. Of these, DNA index and S-phase fraction (SPF) proved to be the most valuable prognostic indices. DNA aneuploidy was associated with a three-fold risk of death as compared to DNA diploidy (P less than 0.0001). The highest risk of death was associated with hypertetraploid (greater than 2.20) DNA index, whereas a tetraploid DNA index (1.80-2.20) was associated with a relatively low risk. The SPF had significant additional prognostic value in both DNA diploid (P = 0.0002) and DNA aneuploid (P = 0.02) tumors. By combining DNA index and SPF the authors defined three types of DNA histograms, which were associated with favorable, intermediate, and poor prognosis of the patients. DNA diploidy together with low (less than 7%) SPF (type I DNA histogram) was associated with very favorable prognosis, whereas DNA aneuploidy with high DNA index (greater than 2.20) or high (greater than 12%) SPF (type III DNA histogram) was related to the worst prognosis with approximately eight-fold relative risk of death. In a Cox multivariate regression analysis the type of DNA histogram was an independent and most powerful prognostic indicator in breast cancer. The other independent factors in the Cox analysis were primary tumor size, nodal status, and progesterone receptor status.

The prognostic impact of DNA content and S-phase fraction (SPF) of tumour cells was studied in 93 patients with primary breast cancer. Aneuploid DNA content and high SPF were clearly associated with poor differentiation state of tumours and absence of steroid, especially progesterone receptors. Aneuploidy and high SPF tended to become more common with increasing primary tumour size, with more extensive nodal involvement and with more advanced stage of the cancer. Patients with diploid tumours had a slightly longer disease-free interval and survival than those with aneuploid tumours, whereas below median SPF as compared to above median SPF was associated with significantly longer (P less than 0.01) relapse-free interval and survival in patients with stage II-III cancer. We conclude that the DNA analysis of tumour cells is a promising method for the estimation of prognosis in breast cancer patients.

We analyzed nuclear DNA content and S-phase fraction (SPF) from 157 paraffin-embedded ovarian tumors by flow cytometry and compared the results with the clinicopathological features of the tumors. DNA aneuploidy was more common and mean SPF was higher in advanced than in early-stage tumors (p less than 0.001). DNA aneuploidy was most often (75%) observed in undifferentiated tumors (World Health Organization classification) and most seldom (30%) in mucinous carcinomas. Mean SPF values ranged from 17.7% in undifferentiated carcinomas to 11.1% in mucinous carcinomas. DNA flow cytometric results correlated better with nuclear than with histological tumor grade. The proportion of DNA-aneuploid tumors increased from 30% in nuclear grade I to 93% in nuclear grade III, and mean SPF increased from 9.9 to 20.2% (p less than 0.001). DNA ploidy and SPF were independently associated with both stage and nuclear grade of the tumor, whereas the differences between the histopathological tumor types virtually disappeared when the groups were adjusted for nuclear grade. On the basis of these clinicopathological correlations, it appears that DNA ploidy and SPF reflect the malignant potential of ovarian tumors and thus complement the routine histopathological evaluation.