Euploid Tumors Research Articles

Mucoepidermoid Carcinoma of Major and Minor Salivary Glands

Forty-three consecutive cases of mucoepidermoid carcinoma of the major and minor salivary glands were studied to evaluate the influence of the following parameters on prognosis: age, sex, location, local extension, mitotic counts, vascular invasion, grade of differentiation, and nuclear DNA ploidy assessed by image cytometry. The age of the patients ranged between 7 and 84 years. Twenty-eight patients were male and 15 female. Thirty-five tumors were in the major salivary glands and 8 in the minor glands of the oral cavity. According to Healey's criteria, 14 tumors were classified as grade 1, 17 as grade 2, and 12 as grade 3 and, according to the 1990 World Health Organization proposal, 17 tumors were classified as low-grade and 26 as high-grade neoplasms. Image cytophotometric DNA analysis revealed 15 euploid and 6 aneuploid tumors. The range of the follow-up study was 3-156 months (mean, 40.7 months). The 5- year actuarial survival for each of the above-cited parameters was: 100% and 59.1 % for patients aged less than 20 and over 55 years, respectively; 49.6% for male and 85.7% for female patients; 61.3% for major gland and 73.3% for minor gland tumors; 100% and 57.5% for intraglandular and extraglandular major salivary gland neo plasms, respectively; 46.2% for cases with high mitotic rate and 83.1% for those with low mitotic rate; 38.7% and 81.6% for tumors respectively with and without vascular invasion; 100% for grade 1 and 64.7% for grade 2; none of the patients with grade 3 neoplasms were alive at 5 years. Survival was of 100% for low-grade and 40.7% for high-grade tumors. Survival was 92.8% for euploid tumors and 61.3% for the aneuploid ones. In conclusion, survival of patients with mucoepidermoid carcinoma is significantly related to the histological grade of differentiation, the mitotic count, the presence of vascular invasion and the DNA ploidy of the tumor cells. Int J Surg Pathol 1(1):3-12, 1993

Breast carcinoma growth rate described by mammographic doubling time and S-phase fraction. Correlations to clinical and histopathologic factors in a screened population.

In a retrospective study, correlations among mammographic doubling times (DT), clinicopathologic prognostic factors, and cytometric predictors were examined. One hundred fifty-eight patients with the possibility to calculate mammographic tumor DT were selected and the tumors were histologically reexamined and flow cytometric analysis for ploidy and S-phase fraction (SPF) was performed. The tumors were Stage I in 68%, and 45% were detected by mammographic screening. DT ranged from 0.6 months to an indefinite time (median, 9.0 months). Short DT was significantly correlated to large tumor size (P = 0.01) and advanced pathologic tumor stage (P = 0.016), but there was no correlation between DT and histologic grade. Ploidy analysis indicated that there were 57% aneuploid and 7% tetraploid tumors. There was a significant overrepresentation of euploid tumors among tumors smaller than 10 mm (P = 0.02). Ploidy was correlated to histologic grade (P less than 0.001) and DT (P = 0.009). SPF was calculated in 122 cases. SPF correlated significantly with pathologic stage (P = 0.002), tumor size (P = 0.037), histologic grade (P = 0.001), the presence of axillary lymph node metastases (P = 0.046), DT (P = 0.02), and DNA ploidy (P less than 0.001). Compared with interval carcinoma, screening-detected carcinoma showed favorable characteristics concerning size, stage, DT, ploidy, and SPF but not regarding histologic grade and axillary lymph node metastases. DT shows great variations. Factors related to tumor biology (i.e., DT, DNA ploidy, and SPF) are strongly correlated with one another, but they have no correlation with axillary lymph node metastases. Cancer detected by screening is discovered at an early stage and shows favorable characteristics concerning DT, ploidy, and SPF.

Prognostic factors in parathyroid cancer: a review of 95 cases.

The clinical course, histopathology, and tumor DNA distribution patterns were analyzed in 95 patients with parathyroid cancer. The median follow-up was 6 years (range 1-25 years). Eighteen patients received a benign diagnosis at their first operation. The initial procedure was tumor resection in 42 patients and tumor resection plus partial or total thyroidectomy in 40 patients. Forty patients developed recurrent disease and 36 patients underwent 1 to 9 re-operations. Cervical recurrence and lung metastases were most commonly encountered. The median time from the first operation to recurrence was 33 months (range 1-228 month). Twenty-one patients died of parathyroid cancer a median of 28 months following discovery of their first recurrence. The histopathological reevaluation confirmed unequivocal parathyroid cancer, i.e., infiltration and/or metastases, in 41 cases. Fifty-four cases lacked these criteria but showed various forms of atypia. Image cytometry demonstrated tumor aneuploidy in 26 of 39 cases with definite cancer by histological criteria, compared to the 13 of the 52 with equivocal histological diagnosis. Twelve patients with aneuploid tumors and 7 patients with euploid tumors died of parathyroid cancer. In a multivariate analysis, patients treated with extensive surgery, i.e., tumor resection and unilateral or bilateral thyroidectomy, had a longer survival and a longer relapse-free period. Other factors of importance for survival were age and histopathology. Histopathology and an aberrant nuclear DNA content were important factors for the time to recurrence. We conclude that histopathology alone is unable to confirm a cancer diagnosis in the absence of infiltration and/or metastases. Because recurrence may occur late, patients should be followed closely. Even repeated surgical interventions have proven beneficial.

DNA cytophotometry and prognosis in ovarian tumors of borderline malignancy. A clinicomorphologic study of 80 cases.

Surgical specimens of 80 ovarian tumors of borderline malignancy (OTBM) were investigated by scanning DNA cytophotometry. Diploid or euploid DNA histograms were found for 21 tumors, whereas 59 OTBM showed noneuploid or aneuploid DNA patterns. All patients were followed-up after surgery for at least 3 years (mean observation period, 6.7 years). Follow-up showed 11 cases of recurrent disease and 6 deaths. DNA findings and several other morphologic and clinical details (including patient age, histologic type and stage of disease, and extent of therapy) were correlated to the postoperative course. Statistical analyses disclosed that, of these parameters, only DNA content significantly affected prognosis. Recurrences and deaths resulting from tumor exclusively were observed among patients with noneuploid or aneuploid OTBM, whereas none of the diploid or euploid tumors recurred (P less than 0.05). DNA cytophotometry thus might be regarded as an effective complementary means to assess the prognosis of individual OTBM cases.

DNA ploidy in squamous oesophageal carcinoma

The role of DNA ploidy in the management of oesophageal carcinoma is unclear. Most studies have employed flow cytometry (FC) for DNA analysis but some have used image analysis (IA) of tissue sections. In this study aneuploidy rates in stage IIa squamous tumours were determined by both FC and IA of cell suspensions and results were compared with outcome in two patient subgroups. Group 1 ( n = 15) were patients who died from tumour recurrence within 1 year of surgery while Group 2 ( n = 21) were patients who survived tumour free for at least 1 year. Aneuploidy rates differed significantly between techniques; 29 of 36 tumours (81%) were aneuploid by IA compared with 19 of 34 (56%) by FC ( P < 0.05). Aneuploidy rates differed significantly between groups 1 and 2 as determined by FC (79% versus 40%) ( p < 0.05) but not by IA (93% versus 71%) ( P = ns). Euploid status was a good prognostic indicator; 6 of 7 (86%) patients with euploid tumours by IA and 12 of 15 (80%) by FC ( P < 0.05) survived more than 1 year. The sensitivity and specificity of euploidy was 93% and 28.6% for IA compared with 78.6% and 60% for FC. Since 33 (92%) of these tumours exhibited a marked peritumoral desmoplastic or chronic inflammatory reaction IA, being more sensitive to subtle nuclear change, may be a more appropriate technique than FC for evaluation of the role of ploidy in such tumours.

Reduced proliferative activity of polyploid cells in primary hepatocellular carcinoma.

The proliferative activity of tumor cells differing in DNA content (ploidy) and nuclearity was investigated in primary hepatocellular carcinomas of the rat by flow cytometric analysis of collagenase-isolated cells immunostained after labelling with bromodeoxyuridine (BrdU) in vivo. The diploid cell fraction in these euploid tumours was higher than in normal liver, and the rate of binucleation as well as the proliferative activity of the binuclear cells was very low. The highest proliferative activity (BrdU labelling index) was found among the diploid tumour cells. The activity in mononuclear tetraploid and octoploid cells was reduced in inverse proportion to their increasing DNA content, possibly suggesting a loss of proliferative potential associated with polyploidization. There was a significant correlation between the proliferative activity of hepatocellular carcinoma cells and nonparenchymal liver cells in the different tumours, indicating that different cell types within a tumour may respond to common growth stimuli. Treatment of tumour-bearing rats with a promoting carcinogen (2-acetylaminofluorene) resulted in significant stimulation of tumour cell proliferation (all ploidy classes), whereas the proliferation of non-parenchymal (stromal) cells in the tumour was slightly inhibited.

Proliferation and DNA ploidy in malignant breast tumors in relation to early oral contraceptive use and early abortions

In 175 premenopausal breast cancer patients, a history of oral contraceptive (OC) use before 20 years of age was significantly associated with higher tumor cell proliferative activity, as indicated by a higher S-phase fraction (SPF), and a higher fraction of DNA aneuploid tumors, compared with later or never users (P = 0.05 and p = 0.01, respectively). The higher SPF among early OC users was apparent in patients with aneuploid tumors but not in patients with euploid tumors. Abortions (spontaneous or induced) before the first full-term pregnancy also were associated with a higher SPF compared with other young patients with breast cancer (P = 0.03). Adjusting for parity and abortions or OC use, respectively, an early OC use was associated with a 43% higher SPF and early abortions were associated with 49% higher SPF. Younger patients had a higher SPF and a higher frequency of aneuploid tumors, but this was found to be because the users of OC had a lower median age at diagnosis. Among never users, no significant age relationship was seen for SPF or the frequency of aneuploidy. For the DNA analyses there is a selection of patients with breast cancer with larger tumors, and therefore the conclusions drawn in this article may not be generalizable to patients with smaller primary tumors, e.g., cases diagnosed at breast cancer screening. The higher tumor proliferative activity and frequency of aneuploidy in early OC users are in line with previously reported findings of worse prognostic indicators and a worse survival in early users of OC compared with other young women with breast cancer.

Prognostic Value of flow cytometric DNA content analysis in granulosa cell tumor of the ovary

The nuclear DNA content and S-phase fraction of 23 ovarian granulosa cell tumors were measured from paraffin-embedded tissue with flow cytometry. Crude survival of the patients with a euploid tumor (17 diploid, one tetraploid) was more favorable than that of the patients with an aneuploid tumor (n = 5, P = 0.02). The percentage of S-phase cells was a good predictor of survival. If more than 6% S-phase cells were present in the DNA histogram, both crude survival (P = 0.0001) and survival corrected for intercurrent deaths (P = 0.0001) were clearly inferior as compared with tumors with less than 6% S-phase cells. The results indicate that DNA flow cytometric study may provide a rapid and valuable method to predict the biological behavior of granulosa cell tumors of the ovary.

Nuclear DNA content and survival in medullary thyroid carcinoma

In a nationwide study of medullary thyroid carcinoma, the relation between nuclear DNA content and survival was studied in 211 patients with complete follow-up of up to 27 years. Morphologically identified tumor cells were analyzed by an image cytometric method. The DNA histograms were classified (1) by an objective method in which the degree of aneuploidy was defined as the percentage number of tumor cells with DNA values exceeding the 90th percentile of diploid control cells (P90), and (2) according to a subjective evaluation of whether the DNA profiles represented a euploid or an aneuploid DNA pattern. Both classifications separated groups with marked differences (P less than 0.001) in survival. A multivariate proportional hazards analysis indicated that each method provided additional information when the other one was taken into account. In patients whose tumors were classified according to the objective P90 method 1, the relative hazards (with 95% confidence interval) in the groups with P90 values of 36% to 69% and greater than or equal to 70% were 1.6 (0.9-2.8) and 2.1 (1.1-4.0) respectively, compared with the reference group. With the subjective method 2 the corresponding figure for the group with aneuploid tumors was 1.7 (1.0-2.0) compared with the group with euploid tumors.

Chromosome studies in thyroid neoplasia.

Cytogenetic studies in thyroid neoplasia were performed by G-banding of chromosome preparations obtained from the in vitro cultures of nine adenomas, one follicular carcinoma, five papillary carcinomas, and two medullary carcinomas. Complex structural chromosome aberrations were found in one adenoma. Two more adenomas, both composed of Hürthle cells, showed multiple numerical chromosome deviations with trisomy 4 and tetrasomy 7 in common. Six metastasizing carcinomas were characterized by normal stemlines, which indicates that malignancy in thyroid neoplasia cannot be excluded by cytogenetic techniques used currently. Comparisons between cytogenetic findings and cytophotometric DNA measurements in the material studied illustrate that euploid tumors represent a heterogenous group including cases with various gross structural chromosome aberrations of yet unknown clinical significance. Further studies of additional material with long-term follow-up are called for by our findings of structural and numerical chromosome aberrations in follicular neoplasms that are benign according to histologic criteria.

Feulgen DNA Values in Wilms' Tumour in Relation to Prognosis

Single-cell DNA measurements were performed on Feulgen-stained nuclei in 4 mu tumour sections from 30 patients with Wilms' tumour, all treated with initial nephrectomy. Fifteen patients died from their tumours within 2 years of nephrectomy. The remaining 15 had survived for at least 10 years without evidence of recurrence. Based on the histograms, the tumours were classified as being euploid (E type) or aneuploid (A type). Twenty tumours were euploid and 10 aneuploid. The euploid tumours predominated in the group of survivors. The group of 10 aneuploid tumours contained 7 tumour deaths. The tendency for aneuploid tumours to be associated with early tumour death, however, was not statistically significant.

Cellular DNA content and metastasis pattern in colorectal carcinomas.

Tissue samples from primary tumours and metastases obtained from surgical specimens and autopsies were investigated by flow cytometry for their DNA content. Of the 149 cases investigated, 124 autopsy cases of colorectal carcinoma were suitable for study. The pattern of metastatic spread in each individual case was analysed with reference to the autopsy records. A subgroup of euploid primary tumours was observed which should be considered separately with regard to its biological behavior, tumour location and extent of metastases. In these tumours, the liver was the first and final organ of haematogenic metastatic spread at an above-random frequency and a grossly metastatic liver with less than 15% of residual normal parenchyma was present at the same time. Furthermore, we observed stem-lines deviating in ploidy from the primary tumour in the metastases of nine cases. This indicates the chromosomal heterogeneity of colorectal carcinomas.

Tumor DNA content as a prognostic feature in advanced epithelial ovarian carcinoma

Tumor DNA content (ploidy) was determined in 84 patients with epithelial ovarian carcinoma, Stage II–IV. A total of 251 DNA histograms generated by flow cytometry on cells derived from paraffin-embedded specimens were analyzed retrospectively. Of the 84 patients, 44 had tumors which were aneuploid, whereas 33 had diploid, and 7 had tetraploid tumors. Cox regression analysis revealed that age ( P < 0.001), stage ( P < 0.001), and ploidy ( P < 0.001) were independent prognostic features. The median survival time was 19 months and 48 months, respectively, in aneuploid and euploid tumors ( P < 0.001). The size of residual after surgery lost its significance when corrected for stage. Multivariate analysis in Stage III tumors revealed that ploidy was the most important prognostic factor ( P < 0.001) followed by age ( P < 0.025). A remarkable stability of cellular DNA content was found when the primary tumor was compared to the following groups: (1) various metastatic specimens from the primary operation in the same patient; (2) specimens analyzed sequentially from primary, secondary, and tertiary exploratory laparotomy; and (3) peritoneal washings before and after intraperitoneal chemotherapy.

Comparison Between Histologic Type, Estrogen Receptor, and Nuclear DNA Content in Mammary Carcinoma

Histological specimens from 80 invasive breast carcinomas comprising typical cases of 16 ductal, nine papillary, 14 comedo, 13 colloid (mucous), 15 lobular, and 13 medullary carcinomas were examined with respect to nuclear DNA and estrogen receptor content. In agreement with previous studies, ductal carcinomas were found to exhibit different types of nuclear DNA distribution patterns, i.e., tumors with DNA values in the normal diploid or tetraploid regions indicative of good prognosis (euploid tumors) or those with values exceeding the normal tetraploid region indicative of poor prognosis (aneuploid tumors). The majority of the papillary and colloid tumors were euploid, while comedocarcinomas in general had aneuploid profiles. These findings are in agreement with expected survival within these patient groups. In lobular breast carcinomas, the correlation between the DNA distribution patterns and expected patient survival was less obvious; and in medullary carcinomas where the vast majority of the tumors showed aneuploid DNA profiles, the correlation to expected patient survival was low. Thus, lobular carcinoma in general seems to have a worse prognosis than is expected from nuclear DNA analysis, whereas medullary carcinomas in general seem to carry a better prognosis than indicated from DNA measurements. In agreement with earlier reports there was a good correlation between nuclear DNA content of the tumor cells and cytosol estrogen receptor values, i.e., euploid tumors in general exhibited relatively high receptor levels, whereas aneuploid tumors had low or unmeasurable estrogen receptor levels.

Flow-cytometric DNA analysis in primary breast carcinomas and clinicopathological correlations.

Flow-cytometric DNA analyses of single cell nuclei were performed on nuclear suspensions prepared from biopsies of primary breast carcinomas in 638 patients. Propidium-iodide-stained cell nuclei were analysed in an Ortho 50-H Cytofluorograph. The patients were staged by the TMN classification. Sixty percent of all the patients had aneuploid primary breast carcinomas; of size T1, T2, T3, and T4 tumors, 51%, 63%, 67%, and 75% were aneuploid, respectively. The proportions of aneuploid tumors in each of the tumor stages SI, SII, SIII, and SIV were 47%, 62%, 67%, and 69%, respectively. This trend to increasing aneuploidy proportions with more advanced disease was significant in contrast to the degree of aneuploidy found in relation to axillary nodal tumor involvement. Multiple aneuploid cell populations were found in 109 (17%) tumors. With a mean follow-up time of 16 months, 92 patients have relapsed out of 540 completely staged patients with unilateral breast cancer with no distant metastases at the time of initial treatment. When the influence of various treatments and tumor stage are not considered, the recurrence rate was twice as high among patients with aneuploid primary tumors than among patients with euploid tumors. The differences in relapsing rates among patients with euploid and aneuploid primary tumors decreased with more advanced disease. Out of 170 patients with T1 tumors, 17 relapsed and 16 of these were aneuploid. No such difference in relapse rate in relation to ploidy was, however, found in patients with more advanced primary disease.

The analysis of malignancy by cell fusion. 3. Hybrids between diploid fibroblasts and other tumour cells.

ABSTRACT Diploid fibroblasts were fused with cells of two highly malignant near euploid tumours, a sarcoma induced by polyoma virus and a spontaneous carcinoma; the resulting hybrid cells were tested for their ability to grow progressively in vivo. In the case of the sarcoma/fibroblast hybrids, 15 clonal populations, each derived from a separate primary fusion, were examined. Most of these populations already showed substantial losses of chromosomes by the time enough cells had been generated to permit chromosomal analysis; but a few clones were isolated with chromosomal constitutions approximating to the sum of the 2 parental chromosome sets. Those clones that had undergone substantial chromosomal losses were highly tumorigenic, but some of the clones that contained the complete, or almost complete, chromosome sets of both parent cells, showed a greatly reduced take incidence. Initially these clones produced very few tumours, but the take incidence rose as continued cultivation of the cells in vitro resulted in progressive loss of chromosomes. In the case of the carcinoma/fibroblast hybrids, clonal populations with very high chromosome numbers were selected for special study. These were also found to have a very low take incidence, comparable to that of the hybrids formed by the fusion of the tumour cells with L cell derivatives. None of the tumours produced by the injection of either the sarcoma/fibroblast or carcinoma/fibroblast hybrids were composed of cells containing chromosome complements corresponding to the sum of the 2 parental chromosome sets; all the tumours were formed by selective overgrowth of cells from which some chromosomes had been eliminated. These results indicate that the malignancy of the tumour cells can be suppressed by diploid fibroblasts as well as by the L cell derivatives, but that the rapid chromosome losses characteristic of hybrids between these tumour cells and diploid fibroblasts generate malignant segregants with much greater frequency. In the range of material examined in the present experiments malignancy thus behaves as if it were a recessive character.