Flow cytometric analysis of DNA ploidy in canine mammary tumors.

DNA ploidy has been determined using flow cytometry in 23 nonmalignant and 34 malignant (primary and metastatic) mammary tumors from 46 dogs. This parameter was compared with clinical stage, histology, and estrogen and progesterone receptor analysis. Twenty-one of 34 cancers (61.8%) from 32 dogs were DNA aneuploid. Aneuploidy was also found in 4 of 23 nonmalignant tumors (17.4%) from 20 dogs. Regional lymph nodes were involved in 6 of 10 diploid and 3 of 9 aneuploid cancers of dogs with operable disease. The aneuploidy incidence was higher in dogs that had distant metastasis at initial diagnosis (8 of 11) than in those presented with local or locoregional disease (9 of 19), although this difference was not statistically significant. DNA aneuploidy incidence was not found to be related to histological tumor type, histological malignancy grade, nuclear grade, or steroid receptor presence. Heterogeneity in DNA content was found in 4 of 32 cancers (30 dogs) in samples from primary or locally recurrent lesions. In 3 of 16 cancers that were analyzed both at the primary and at secondary sites of growth, a significant variation in DNA content was observed. The degree of aneuploidy in the dog cancers was much lower than seen for human breast carcinomas with a relatively high frequency of hypoploid stemlines (7 of 34 cancers, 20.6%). The frequency distribution of DNA indices in dog mammary cancers indicates that aneuploidy evolution probably differs from that of human breast cancer.

Flow cytometric DNA ploidy analysis of feline mammary tumors.

Flow cytometric DNA analysis was performed on biopsies from 9 nonmalignant and 111 malignant (primary and metastatic) feline mammary lesions. In our series, 46.3% of the primary mammary carcinomas appeared to be aneuploid, whereas all but one benign breast lesion were diploid. The degree of aneuploidy in carcinomas was low, with a relatively high number of primary tumors (12 of 82) displaying hypodiploidy. Aneuploidy was not found to be correlated with any specific histological tumor type, vascular invasion, tumor size, or histological malignancy grade or with the separate components thereof. Comparison of the ploidy in primary and metastatic tumors from the same cases revealed a remarkable stability, both in time and location of appearance of the metastases. It is concluded that with respect to DNA ploidy feline mammary carcinoma has more in common with canine mammary carcinoma than with human mammary carcinoma. Further prospective studies are necessary to clarify the implications of aneuploidy in feline mammary carcinoma for tumor behavior and prognosis.

DNA ploidy and cell kinetic characteristics in canine non-Hodgkin's lymphoma.

Malignant lymphoma in the dog is frequently postulated and used as a therapeutic model for non-Hodgkin's lymphoma (NHL) in humans. In this study DNA ploidy and the cell kinetic characteristics of canine malignant lymphoma were studied by flow cytometric (FCM) nuclear DNA measurements on fresh frozen tumor tissue from 94 dogs with NHL and on material from non-neoplastic lymph nodes from 20 dogs. The results were correlated with histomorphology, immunophenotype and survival. All non-neoplastic tissues were diploid, whereas of the 94 lymphomas 74 were diploid or near-diploid and 20 aneuploid. Of the aneuploid lymphomas, 1 contained a hypoploid cell population. DNA-indices of the aneuploid peaks ranged from 0.87 to 1.21 (mean 1.11). The mean S-phase fraction (8.2%, SD 4.8) was significantly lower in the non-neoplastic tissues than in the lymphomas (11.4%, SD 5.1). A linear correlation was observed between FCM S-phase fractions and bromodeoxyuridine (BrdU) labeling indices (r = 0.78; p < 0.001) determined in paraffin-embedded tissue sections from 18 dogs with NHL after in vivo BrdU labeling. DNA ploidy status did not correlate to the S-phase fraction. There were no differences in S-phase fraction and DNA ploidy between B cell and T cell lymphomas or between different histological classes using the Working Formulation. No correlation was found between S-phase fraction or DNA ploidy and survival in a series of 59 dogs treated with a combination chemotherapy protocol. It is concluded that the frequency of DNA aneuploidy in canine malignant lymphoma is similar to that in human NHL. In contrast to findings in human NHL, however, no relationship was found between DNA ploidy or cell kinetic features and histomorphology or prognosis.

Paragangliomas of the head and neck region show complete loss of heterozygosity at 11q22-q23 in chief cells and the flow-sorted DNA aneuploid fraction.

Nonchromaffin paragangliomas of the head and neck region, also known as glomus tumors, are usually benign neoplasms consisting of clusters of chief cells surrounded by sustentacular cells arranged in so-called 'Zellballen.' Most of the patients have a familial background. In a previous study, examining all chromosome arms, we found loss of heterozygosity (LOH) predominantly at the chromosome 11q22-q23 region, where the disease causing gene PGL1 has been located by linkage analysis. However, all tumors showed only partial loss of allele signal intensities, and it was not clear whether this represented allelic imbalance or cellular heterogeneity. In the current study, we have performed LOH analysis for the 11q22-q23 region on DNA-aneuploid tumor cells, enriched by flow sorting, and on purified chief cell fractions obtained by single-cell microdissection. Complete LOH was found for two markers (D11S560 and CD3D) in the flow-sorted aneuploid fractions, whereas no LOH was found in the diploid fractions of three tumors. The microdissected chief cells from two of these tumors also showed complete LOH for both markers, indicating that the chief cells are clonal proliferated tumor cells. These results indicate that the PGL1 gene is likely to be a tumor suppressor gene, which is inactivated according to the two-hit model of Knudson. Furthermore, it shows that chief cells are a major if not the sole neoplastic component of paragangliomas.

Molecular identification of a partial hydatidiform mole.

Specimens of a vacuum curettage were microscopically indicated for a hydatidiform mole. The combination of three different approaches identified the specimen as a partial mole caused by the fertilization of a haploid ovum by sperm containing a haploid or diploid nucleus with one or two sets of paternal genetic material. Interphase fluorescence in situ hybridization identified three chromosome 1 centromeres, and DNA flow cytometry revealed a peak with a DNA index of 1.50. The combination of flow cytometric cell sorting and microsatellite marker polymerase chain reaction proved that in this case two alleles were from paternal origin. Because it is known that partial hydatidiform moles have a tendency for recurrence, specimens from the same patient of an earlier executed vacuum curettage were investigated. Microdissection of the villi was performed before DNA isolation in this case as too few villi were present for DNA flow cytometry and cell sorting. In this case, no evidence was fond for additional alleles. This study shows the diagnostic potential of microsatellite markers for genetic typing of hydatidiform moles.

The (iso)enzyme activities of lactate dehydrogenase, alpha-hydroxybutyrate dehydrogenase, creatine kinase and aspartate aminotransferase in human myocardial biopsies and autopsies.

In a study on enzyme activities in normal human myocardial tissue the activities of lactate dehydrogenase (LDH), alpha-hydroxybutyrate dehydrogenase (HBDH), creatine kinase (CK), and aspartate aminotransferase (AST), and the activities of the isoenzymes of LDH (1--5) and AST (c and m) were measured in myocardial specimens obtained from live patients (18 biopsies) and postmortem (14 autopsies). Comparison of measurements in 4 right ventricular biopsies from 1 patient with those in 9 right ventricular biopsies from 9 patients shows that (1) the enzymes studied are homogeneously distributed throughout the right ventricular myocardium, (2) errors in the measurement of the activities of the enzymes studied are within 5%, and (3) inter-individual differences in myocardial enzyme activities are quite considerable (10--17%). No significant differences in the activities of the enzymes studied were found between left and right ventricular myocardium. The effect of autolysis on the activities of the enzymes studied in myocardial specimens obtained postmortem is small, amounting to -1% to -6% in the first 10 h, irrespective of whether a linear or an exponential decay of enzyme activity is assumed. Comparison of myocardial enzyme activities in biopsies with those in autopsies, the latter being corrected for autolysis-induced inactivation, reveals significant differences with respect to CK, AST and mAST. At present it is impossible to conclude whether these differences were due to the different conditions existing before the myocardial specimens were frozen at -20 degrees C or to the different types of pathology present in the hearts from which the specimens were taken.

Pyridoxal-5'-phosphate-induced stimulation of aspartate aminotransferase and its isoenzymes in human myocardial biopsies and autopsies.

The extent to which pyridoxal-5'-phosphate stimulates the activities of aspartate aminotransferase and its cytoplasmic and mitochondrial isoenzymes was measured in six human left ventricular biopsies obtained freshly during open-heart surgery, and in 13 human left ventricular autopsies. A concentration of pyridoxal-5'-phosphate of 15 mumol/l for 1 h is sufficient to convert any apoenzyme to holo-enzyme. Pyridoxal-5'-phosphate-induced stimulation of aspartate aminotransferase is 26 +/- 6% (+/- SD) in myocardial biopsies (range 22-34%) and 112 +/- 54% in myocardial autopsies (range 35-200%). The extent of stimulation of the cytoplasmic and mitochondrial isoenzymes is 18 +/- 9% and 32 +/- 6%, respectively, in myocardial biopsies, and 150 +/- 57% and 100 +/- 62%, respectively, in myocardial autopsies. The greater extent and variation of the pyridoxal-5'-phosphate-induced stimulation of aspartate aminotransferase and its isoenzymes in myocardial autopsies compared to that in myocardial biopsies is caused by autolysis and its duration. Autolysis depresses myocardial aspartate aminotransferase activity measured in the absence of pyridoxal-5'-phosphate which effect is more prominent for the cytoplasmic than for the mitochondrial isoenzyme.

The difference in DNA ploidy pattern between some canine and human neoplasms appears to be genuine and a reflection of dissimilarities in DNA aneuploidy evolution.

In a reaction to the article by Deitch et al, (Anticancer Res 13: 2117-2118, 1993) evidence is presented that flow cytometrically detected DNA-hypodiploidy in canine neoplasms is genuine and not an artefact caused by autolysis or chemotherapy. Intervals between removal of tumors and freezing in our studies were much shorter (average 15 min, maximum 30 min) than e.g. for human breast tumors in which the percentage of hypodiploidy is about 2%. Also average CVs for the G0, 1 peaks in our FCM analysis of canine tumors (mammary 2.27 + 0.06, n = 179); thyroid 2.57 + 0.13, n = 88) were equal to or less than those usually found in the comparable human tumors. Biological arguments in favor of the existence of genuine hypodiploid stemlines are the finding of tetraploidized subclones of the original hypodiploid clone, the reappearance of the same hypodiploid stemline in distant metastases during clinical follow up, and the isolation of a cytogenetically and flow cytometrically hypodiploid cell line from a primary canine mammary carcinoma. It is concluded that Deitch et al, incorrectly have invoked autolysis as a source of hypodiploidy in our original studies on canine neoplasms. Our evidence for interspecies differences in the evolution of aneuploidy in tumors of the same organ therefore remains unchallenged.

Flow-cytometric DNA ploidy analysis in primary and metastatic canine thyroid carcinomas.

DNA ploidy was measured by flow cytometry in 36 primary malignant thyroid neoplasms (including 6 bilateral tumours which were considered as separate neoplasms) from 30 dogs. In addition, DNA ploidy was determined in local recurrences in 3 dogs, and in 18 metastatic sites from 14 dogs. Aneuploidy was found in 21 of 36 (58%) primary sites. Eighteen of the 21 (86%) aneuploid tumours contained hypodiploid cell populations, with 12 having single hypodiploid peaks, and 6 being multiploid. Three other tumours had single aneuploid peaks with a DNA index (DI) greater than 1.0. The DIs in local recurrences were identical to those in the original neoplasms. Ploidy status (diploid vs. aneuploid) was identical in primary and metastatic sites in 10 out of the 14 dogs. Aneuploidy was more frequent in carcinomas from dogs with distant metastases (78%) than from dogs with less advanced stages of disease (53%), although this difference was not significant. There was no significant correlation between DNA ploidy and histopathological variables. From the strikingly high frequency of hypodiploidy in canine tumours, it is concluded that ploidy evolution in canine neoplasms may differ from that in human tumours.

DNA flow cytometry of canine mammary tumours: the relationship of DNA ploidy and S-phase fraction to clinical and histological features.

The DNA ploidy status and S-phase fraction (SPF) of benign proliferative lesions (BPL) and malignant tumours (MT) in the mammary glands of dogs were determined by flow cytometric analysis and the results were related to their clinical and histological features. Seven (14.3 per cent) of 49 BPL and 16 (48.5 per cent) of 33 primary MT had aneuploid G0,1 peaks (P < 0.001). Hypodiploid G0,1 peaks were found in one BPL and in five primary MT. The DNA ploidy status of primary MT was not found to be associated with their size, nodal status, grade of histological malignancy or nuclear grade. In several cases there was intra-tumour heterogeneity in ploidy status independent of histological heterogeneity. The SPF was significantly higher in 27 primary MT than in 45 BPL when diploid and aneploid cases were combined for comparison (P < 0.05), but not when only diploid cases were compared. Among the primary MT the SPF was higher in aneuploid than in diploid tumours (P < 0.05) and it was higher in five MT from five dogs with regional disease than in 22 MT from 19 dogs with local disease (P < 0.05). The SPF was positively correlated with the grade of histological malignancy (P < 0.05) but not with nuclear grade.

Analysis of tumor heterogeneity in a patient with synchronously occurring female genital tract malignancies by DNA flow cytometry, DNA fingerprinting, and immunohistochemistry.

A case of a patient with bilateral ovarian cancer and a uterine malignant mesodermal mixed tumor with ascites and metastatic disease is presented. Flow cytometry, DNA fingerprinting, and immunohistochemistry were performed to assess the origin of these malignancies. Ploidy analysis showed that both ovarian tumors had different aneuploid stemlines (DNA index [DI] = 1.64, 1.85, right ovary and DI = 1.73, left ovary) indicating independent origins. One of the stemlines in the right ovary (DI = 1.64) was also present in the ascites cells, whereas omentum metastases showed the same stemline (DI = 1.73) as the left ovarian tumor. The uterine malignancy contained three aneuploid stemlines. The highest stemline was associated with epithelial differentiation, but a metastatic origin from the left ovarian tumor seems unlikely. DNA fingerprinting analysis revealed a common change in restriction fragment length pattern in the DNA from all tumor localizations as compared with the patient's constitutional DNA. These results indicate that DNA flow cytometry can be helpful in discriminating intragenital metastatic disease from multiple primary tumors.

Myocardial (iso)enzyme activities, DNA concentration and nuclear polyploidy in hearts of patients operated upon for congenital heart disease, and in normal and hypertrophic adult human hearts at autopsy.

To investigate biochemical characteristics of hypertrophic myocardium of young and adult humans, we analysed myocardial biopsies obtained from 28 mainly young patients undergoing cardiac surgery for congenital heart disease and 41 autopsied hearts from 18 adult normal and 23 hypertrophic human subjects. Myocardial activities of the enzymes creatine kinase and lactate dehydrogenase were independent of age during childhood, but decreased significantly with hypertrophy at adult age. Myocyte nuclei showed increased polyploidization during childhood which was progressive with age, and in the adult stage polyploidization was correlated with heart weight. Nevertheless myocardial DNA concentration fell under both conditions, which is to be ascribed to the 'diluting' effect of myocyte hypertrophy. Before an age of 8 years DNA concentration in the child heart material studied has reached the value found in adult nonhypertrophic hearts, although at that time polyploidization of myocyte nuclei in child hearts was only half the value found in adult non-hypertrophic hearts. Biochemical measurement of DNA concentration in peroperatively taken myocardial biopsies may contribute to the in vivo diagnosis of ventricular hypertrophy in quantitative terms, in combination with radiology, echocardiography and histology.

Molecular genetic analysis of flow-sorted ovarian tumour cells: improved detection of loss of heterozygosity.

Detection of loss of heterozygosity (LOH) is usually performed on homogenised tumour specimens. In this type of analysis samples with a low percentage of tumour cells have to be excluded and possible intra-tumour heterogeneity is obscured. In this study we report the application of polymerase chain reaction (PCR)-driven LOH detection with in total 22 microsatellite markers for chromosome 1q, 3p, 3q, 4p, 6p, 6q, 11p, 11q, 17p, 17q, 18p, 18q, Xp and Xq on flow-sorted cells from fresh and paraffin-embedded ovarian tumour tissue. Titration experiments showed that LOH can be detected with as few as 100 cell equivalents of DNA. Clear examples of LOH could be detected in the sorted aneuploid fractions from one unilateral and two bilateral ovarian tumours from three patients. In two samples the sorted fraction was less than 10% of the total sample. The bilateral tumours from the same patient showed loss of identical alleles for one marker (case OV64) and two markers (case OV69), indicative of their monoclonal origin. Multiparameter flow cytometry using two different ovarian tumour markers (MOv18 and BMA180), an anti-cytokeratin monoclonal antibody (MAb) (M9), an anti-vimentin MAb (V9) and a MAb against the panepithelial antigen 17-1A on the fresh ascites cells of the fourth ovarian cancer patient was used to investigate possible intra-tumour heterogeneity. We showed the presence of at least three phenotypically different populations, of which the diploid, keratin-positive, vimentin-negative population showed a similar LOH pattern as the aneuploid population (DNA index = 1.7), indicative of its neoplastic origin. The same LOH pattern was shown in an omentum metastasis from this patient also having the same aneuploid DNA index of 1.7. The sharing of the same LOH pattern by the diploid and aneuploid tumour cell populations suggests that the observed allele loss events occurred before the development of aneuploidy. PCR on flow-sorted cells is thus an important tool to study clonal diversity in tumours.

Flow cytometric DNA-ploidy analysis of synchronously occurring multiple malignant tumors of the female genital tract.

In this study the authors applied flow cytometric DNA-ploidy analysis to multiple female genital tract malignant tumors in 43 patients, most of whom (n = 37) had bilateral ovarian cancer. An algorithm was developed for calculation of the likelihood ratio of the probabilities that measured DNA index differences between multiple tumor localizations within the same patient could be attributed to measurement variation or to true biologic DNA content differences. The results of this statistical analysis show that in 72% of the cases (31 of 43) this probability ratio exceeded 1. Because the probability that two independent tumors will have a near-identical aneuploid DNA content is very low, this finding supports a metastatic process rather than the occurrence of multiple primary tumors in these patients. Thus, flow cytometric DNA-ploidy analysis can be helpful in the identification of metastatic disease in patients with multiple female genital tract malignant tumors.

Evidence of clonal divergence in colorectal carcinoma.

BACKGROUND: The aim of this study was to investigate the generation of DNA ploidy diversity in different stages of colorectal carcinoma development. METHODS: DNA flow cytometry was performed on tissue samples from 20 colorectal adenomas, 38 colorectal carcinomas, 30 lymph node metastases, and 70 hematogenous metastases. RESULTS: DNA aneuploidy was detected in 30% of the adenomas, 82% of the primary colorectal tumors, 57% of the lymph node metastases, 92% of the liver metastases, and 100% of the other distant hematogenous metastases. Multiple DNA tumor stemlines were found in 10%, 39%, 29%, 24%, and 40%, respectively. Sixty-two percent of the DNA tumor stemlines detected in the lymph node or liver metastases were also present in the primary tumors. In primary carcinomas and lymph node metastases, the DNA index distribution had a bimodal shape with a minimum at the 1.2-1.4 region. In the hematogenous metastases, a higher percentage of hypertetraploid stemlines was found. CONCLUSIONS: The emergence of DNA aneuploidy as well as clonal divergence seems to take place during the transition from adenoma to carcinoma. The DNA aneuploid stemlines formed during this phase remain relatively stable over time, although ongoing clonal evolution at distant metastatic tumor sites cannot be completely ruled out.

Frequency and degree of aneuploidy in benign and malignant thyroid neoplasms.

The frequency and degree of aneuploidy in 44 benign and 124 malignant thyroid neoplasms were analyzed by DNA flow cytometry. Single aneuploid cell populations were found in 72% of the undifferentiated carcinomas, 64% of the follicular carcinomas, 24% of the papillary carcinomas and in 24% of the follicular adenomas. Multiple aneuploid cell populations were detected in 4% of the papillary and in 36% of the follicular carcinomas but not in undifferentiated carcinomas. A low degree of aneuploidy was found in well differentiated papillary carcinomas (mean DNA index of aneuploid populations: DI = 1.17; SD +/- 0.09). Significantly higher values were found for aneuploid moderately differentiated papillary carcinomas (DI = 1.46; SD +/- 0.29), well and moderately differentiated follicular carcinomas (DI = 1.61; SD +/- 0.33 and DI = 1.60; SD +/- 0.30, respectively) and undifferentiated carcinomas (DI = 1.72; SD +/- 0.19). High DNA indices were also found in several follicular adenomas (DI = 1.49; SD +/- 0.22). Comparison of the 10-year survival rates of patients with moderately versus well differentiated papillary carcinoma (79 vs. 98 months, respectively) indicates that loss of differentiation and progression of aneuploidy in this tumour type is associated with more aggressive clinical behaviour. Similarly, the high frequency and degree of aneuploidy in undifferentiated carcinomas is in agreement with the very poor survival rate (0% at 10 years) in this group of patients. However, the occurrence of highly aneuploid adenomas and (near)-diploid undifferentiated carcinomas does not point to a direct causal relationship between DNA-ploidy changes and clinical behaviour of these thyroid tumours.

Molecular genetic evidence for unifocal origin of advanced epithelial ovarian cancer and for minor clonal divergence.

Detection of loss of heterozygosity (LOH) and DNA flow cytometry (FCM) were used to trace the origin of bilateral ovarian cancer from 16 patients. From each tumour the DNA index (DI) and LOH patterns for chromosomes 1, 3, 6, 11, 17, 18, 22 and X were determined with 36 microsatellite markers. Formalin-fixed, paraffin-embedded as well as frozen specimens were used. Flow cytometric cell sorting was used to enrich tumour cells for polymerase chain reaction (PCR)-driven LOH analysis. Analysis of the LOH data showed that in 12 of the 16 cases concordance was observed for all informative markers, namely retention of heterozygosity (ROH) or loss of identical alleles in both tumour samples. In four cases discordant LOH patterns were observed. In two cases the discordant LOH was found for one of the chromosomes tested while other LOH patterns clearly indicated a unifocal origin. This suggests limited clonal divergence. In the other two cases all LOH patterns were discordant, most likely indicating an independent origin. The number of chromosomes showing LOH ranged from 0 to 6. Comparison of DNA FCM and the LOH data showed that the latter technique has a higher sensitivity for the detection of a unifocal origin. In 14/16 cases evidence was found for a unifocal origin, while in two cases clonal divergence was found at LOH level and in two other cases clonal divergence at DNA ploidy level. In 12 cases the complete observed allelotype had developed before the formation of metastases, including the two cases showing a large DNA ploidy difference.

At least two different regions are involved in allelic imbalance on chromosome arm 16q in breast cancer.

Loss of heterozygosity (LOH) or allelic imbalance, the latter term referring to both loss and gain of an allele, on the long arm of chromosome 16 has been repeatedly found in cancers of, e.g., the breast and prostate. This indicates the presence of one or more tumor suppressor genes on 16q. To locate the gene(s) more precisely, a detailed allelic imbalance map of 20 polymorphic markers on this chromosome arm was made for 79 sporadic breast carcinomas. LOH of one or more markers was found in 63% of the tumors. Some had allelic imbalance on a region of 16q which failed to overlap with the LOH in other tumors. We therefore assigned two separate "smallest regions of overlap" to 16q and suggest that this chromosome arm contains at least two different tumor suppressor genes.

Loss of heterozygosity and DNA ploidy point to a diverging genetic mechanism in the origin of peripheral and central chondrosarcoma.

Chondrosarcomas are malignant cartilaginous tumors arising centrally in bone (central chondrosarcoma), or secondarily within the cartilaginous cap of a hereditary or sporadic exostosis (peripheral chondrosarcoma). Loss of heterozygosity (LOH) was studied by microsatellite analysis at the loci harboring the EXT genes (implicated in hereditary multiple exostoses), the EXT-like genes, and at 9p21, 13q14, 17p13, and chromosome 10. Nineteen of 20 peripheral chondrosarcomas showed LOH at all loci tested, while only 3 of 12 central chondrosarcomas exhibited LOH, restricted to 9p21, 10, 13q14, and 17p13. LOH at 9p21 did not appear to involve the CDKN2A gene, as assessed by SSCP analysis. DNA flow cytometry demonstrated a wide variation in the ploidy status in peripheral chondrosarcomas (DNA indexes, 0.56-2.01), whereas central chondrosarcomas were predominantly peridiploid. Near-haploidy found in peripheral chondrosarcomas could explain part of the high LOH percentages. Ki-67 immunohistochemistry suggested a higher proliferation rate in peripheral chondrosarcomas. Our results indicate that peripheral chondrosarcomas, arising secondarily to an exostosis, may obtain genetic alterations during malignant transformation, with subsequent genetic instability as demonstrated by a high percentage of LOH and a wide variation in ploidy status. In contrast, peridiploidy and a low percentage of LOH in central tumors suggest that a different oncogenic molecular mechanism may be operative.