Cytogenetic study of ten carcinomas of the bladder: involvement of chromosomes 1 and 11

In direct preparations of ten untreated transitional cell carcinomas of the bladder, chromosomes #1 and #11 were most frequently involved in structural changes (in at least seven tumors each). Three tumors had one or two 11p- chromosomes, and, in other tumors, chromosome #11 had taken part in translocations or isochromosome formation, which, except in one tumor, resulted in a loss of short arm material. Also, there was a tendency for the presence of fewer than expected normal chromosomes #11. Chromosome #1 anomalies are common in most types of tumor; however, chromosome #11 abnormalities, particularly the loss of short arm material, are not common and may thus characterize carcinoma of the bladder, a finding that is of interest in view of the location of an oncogene, c-Ha-ras1, on 11p. Translocations probably involved chromosome #17 in four tumors. Structurally changed chromosomes #3 were seen in four tumors, including one or two 3q- chromosomes in two or possibly three tumors.     

A possible specific chromosome change in transitional cell carcinoma of the bladder

Chromosome changes were ascertained in nine tumor samples from seven untreated patients with transitional cell carcinoma of the urinary bladder. All tumors analyzed showed abnormal karyotypes. In one tumor, a single numerical abnormality (+7) was the sole detectable change. From 1 to 19 structurally abnormal chromosomes could be identified in the remaining 8 tumors. The same abnormality, an isochromosome of the short arm of chromosome #5, was found in five tumors from four patients. We have previously described the presence of this marker chromosome in three of nine cases of transitional cell carcinoma of the bladder. We therefore conclude that i(5p) constitutes the most consistent nonrandom chromosome abnormality in this malignancy. Other chromosomes most frequently involved in structural changes in the present series of tumors were chromosomes #1, #6, #11, and #13.     

Karyotype evolution in a case of chronic myelogenous leukemia with an unusual Philadelphia chromosome translocation,t(4;22), and an additional translocation,t(3;5)

A patient with chronic myelogenous leukemia was found, at the time of diagnosis, to have an unusual Philadelphia chromosome translocation, t(4;22) (q35;q11) and an additional previously unreported translocation, t(3;5) (q27;q22). The blastic crisis, which occurred after 14 months, was characterized by the appearance of i(17q). Ten months later, two different hyperdiploid cell lines with 50 chromosomes were found in about 20% of the metaphases examined.     

Evaluation of chromosome 8 and 11 aneuploidies in washings and biopsy materials of bladder transitional cell carcinoma

We compared chromosome 8 and 11 aneuploidies on bladder biopsy tumor tissues and bladder washing samples of transitional cell carcinoma (TCC) and their relationship to tumor malignancy. Interphase fluorescence in situ hybridization (FISH) was applied to nuclei of washing material and biopsy samples of 17 patients with TCC. Incidence of cells having aneuploidy was clearly nonrandom from patient to patient. There was no significant difference in the incidence of aneuploid frequency for chromosomes 8 and 11 between biopsies of bladder tumors and bladder washing samples (P > 0.05). For chromosome 8, incidence of disomic cells (having two signals) in grade III tumors was significantly lower than in grade II tumors of both washing samples (P = 0.004) and biopsy materials (P = 0.005), indicating a high frequency of aneuploidy. The incidence of nuclei with four or more than four signals of chromosome 8 was significantly higher in grade III tumors than in grade II tumors in washing samples (P = 0.031 and 0.003, respectively). Similarly, in biopsy material, the incidence of nuclei with more than four signals of chromosome 8 was significantly higher in grade III tumors than in grade II tumors (P = 0.004). For chromosome 11, in both washing samples and biopsy materials, the incidence of disomic cells (having two signals) in grade III tumors was significantly lower than that detected in grade II tumors (P = 0.031 and 0.014, respectively), indicating a high frequency of aneuploidy. In biopsy materials, the incidence of nuclei with three or four signals was significantly higher than that in grade II tumors (P = 0.014 and 0.012, respectively). These findings suggest that FISH analysis of bladder washing samples can be effectively detected as genetic changes of bladder tumors. It might predict genetic progression of these tumors, which might be related to tumor stage, because higher stages of tumors showed a higher incidence of aneuploidies of chromosomes 8 and 11.     

Detection of genetic alterations in primary bladder carcinoma with dual-color and multiplex fluorescence in situ hybridization

Cytogenetic studies of bladder cancer have shown several nonrandom aberrations. Numerical aberrations of both sex chromosomes were investigated in 32 primary bladder tumors with bicolor fluorescence in situ hybridization (FISH). Loss of chromosome Y and overrepresentation of chromosome X were observed in subgroups of male patients. Chromosome X was represented normally in female patients. Two of the above primary bladder tumors, a transitional cell carcinoma (TCC) and an adenocarcinoma, were further analyzed with both multiplex FISH (24-color M-FISH) and G-banding. Both cases exhibited 1) common breakpoints on 5q11 approximately q12 and 15q24; 2) involvement of the pericentromeric area of chromosome 13; 3) structural abnormalities of chromosomes 8 and 17, with loss of material on the short arm; 4) structural abnormalities involving chromosome 11; and 5) loss of chromosome Y. The TCC case also exhibited structural abnormalities of chromosome 9, resulting in loss of 9q. The combined G-banding and M-FISH findings could help reveal regions potentially involved in bladder tumorigenesis.     

Nonrandom chromosome changes in carcinoma of the cervix uteri. I. Nine near-diploid tumors

Chromosome analysis of G- and C-banded preparations of tumor material, processed by a direct method, from nine primary carcinomas of the cervix with modal chromosome numbers in the range 41–49 showed the nonrandom involvement of certain chromosomes in structural and numerical changes. Besides chromosome No. 1 (six tumors), structural changes involved chromosomes No. 11 (five tumors with variable breakpoints and translocation partners), No. 3 (three tumors) No. 6 (three tumors), and No. 17 (17p+ in two tumors). A small metacentric (present in duplicate in most metaphases of one tumor), which may have been a 5q? (?) (with an interstitial long arm deletion), was seen in five tumors. Additional normal chromosomes included chromosomes No. 1 (one tumor without structural changes in this chromosome showing trisomy 1) and No. 3 (four tumors showing trisomy in the absence of structural changes involving this chromosome). Losses commonly affected the B, D, and G groups, particularly chromosomes No. 13 (three tumors) and No. 21 (six tumors), as well as chromosome No. 18 (four tumors). Two X chromosomes were present in all tumors except the two with the lowest modal numbers, both of which lacked an X chromosome.     

Chromosome number 11 and Wilms' tumor

Cytogenetic studies have been carried out on cells derived from two Wilms' tumors in vitro. Both tumors had a diploid chromosome range. One tumor was shown to have a definite stemline; 46,XY,4p+,del(9)(q22),11p?q?,11p+, and the other a range of variation chiefly involving chromosomes No. 11, 4, 7, and 2; most changes in chromosome No. 11 took the form of deletions of the short arm.High resolution chromosome analysis of peripheral blood lymphocytes of the two patients revealed apparently normal karyotypes. These findings suggest that changes in the short arm of chromosome No. 11 are important in the development of Wilms' tumor in normal individuals. This association is reinforced by the fact that patients with spontaneous aniridia with a 1 in 3 risk of Wilms' tumor have been reported as having a specific 11p13 deletion.     

Cytogenetic analysis of 62 transitional cell bladder carcinomas

Chromosome analysis of biopsy material obtained after vinblastine pretreatment was carried out in 108 specimens from 89 patients with transitional cell carcinoma of the urinary bladder. Analyzable metaphases were obtained in 62 tumors, but only in nine tumors could karyotypes be analyzed by banding; a conventional technique was used in all others. Ploidy and occurrence of markers corresponded with tumor morphology and invasion and sometimes aided in the clinical evaluation; chromosome anomalies specific for bladder cancer were not revealed. In noninvasive tumors of WHO grade 1 and 2, near-diploid karyotypes with occasional marker chromosomes dominated. Grade 3 tumors showed a variety of grossly aneuploid karyotypes, with an almost constant occurrence of different markers. Superficially invasive G2 tumors had moderately pronounced aberrations with more deviations than noninvasive tumors but without the great variety of G3 tumors.     

Nonrandom karyotypic changes in immortal and tumorigenic Syrian hamster cells induced by diethylstilbestrol

Treatment of Syrian hamster embryo cells with diethylstilbestrol (DES) resulted in the induction of immortal cell lines that progressed and formed tumors in nude mice. Four independently treated cell lines were analyzed cytogenetically at several passages during neoplastic progression. The immortal cell lines at the early passages had no structural abnormalities but did have numerical changes. For example, gain of chromosome 11 was found in all immortal cell lines, and gain of chromosome 19 was found in two of four cell lines. Tumorigenic cells showed not only a variety of numerical abnormalities but also structural abnormalities. Loss of a sex chromosome and gain of chromosome 19 were found in six of seven tumors. Gain of chromosome 11, which was found in all immortal cell lines, disappeared in five of seven tumors. Structural abnormalities involving chromosomes 2 and 3 were found in three of seven tumors. Many marker chromosomes were also found in the tumors. These results support our hypothesis that DES-induced nondisjunction is important in its ability to induce cell transformation and suggests that gain of chromosome 11 and/or 19 may play a role in DES-induced neoplastic progression. Furthermore, these results indicate that for the acquisition of tumorigenicity, additional numerical or structural changes are needed, suggesting that multiple genetic events are required in the multistep process of carcinogenesis.     

Chromosomal analysis of bladder cancer. III. Nonrandom alterations

Chromosome analysis using G- and C-banding was performed on 13 primary transitional cell carcinomas of the bladder. The chromosome preparations were obtained by a direct method. In eight tumors with a (near) diploid modal chromosome number, the most frequently observed chromosome aberrations were: (partial) monosomy 9 in four cases, deletion of 10q in two cases, and partial trisomy 1 in two cases. In five tumors with a modal chromosome number in the triploid or tetraploid range the chromosomes #1, #3, #7, #9, #11, and #17 were numerically and or structurally abnormal in at least four cases. In three out of ten males, the Y chromosome was missing. These findings suggest that the loss of chromosome #9, and possibly also loss of 10q is a primary event in the karyotypic evolution of transitional cell carcinoma of the bladder.     

Near-triploid Ph-positive leukemia

Tetraploid populations have been observed in various types of leukemia, but relatively few reports exist of triploid cell populations in acute or chronic leukemia. We report two cases of Ph-positive leukemia with a modal triploid cell population. Examination of peripheral blood from a 3-year-old boy with Ph-positive acute lymphoblastic leukemia (ALL) and a 68-year-old male with Ph-positive chronic myelocytic leukemia (CML) in blastic crisis revealed modal populations of 72 and 63 chromosomes, respectively. G-banding analysis of both cases revealed the following: karyotypic instability (no clonality), dominant trisomy, and the random association of the Ph chromosome with gains and losses of chromosomes involved in this translocation. The cytogenetic evidence obtained suggests that the triploid cell populations were not derived from a duplication of a hypodiploid cell population, but resulted from random loss of chromosomes from tetraploid cell populations derived from duplication of pseudodiploid cells.     

Clonality and genetic divergence in multifocal low-grade superficial urothelial carcinoma as determined by chromosome 9 and p53 deletion analysis

Multifocality and recurrence are clinically important features of urothelial carcinomas of the urinary bladder. Recent molecular genetic studies have suggested that multifocal urothelial carcinomas are monoclonally derived from an identical transformed progenitor cell. However, most of these studies investigated advanced and poorly differentiated tumors. The study presented focuses on early papillary tumors, including 52 superficial well-differentiated multifocal and recurrent bladder carcinomas from 10 patients. Microdissection separating urothelium from stromal cells was considered essential to obtain pure tumor cell populations. Genetic analysis was carried out by applying two different methods. Dual color fluorescence in situ hybridization (FISH) with centromeric probes for chromosomes 9 and 17 and gene-specific probes for chromosome loci 9q22, 9p21, and 17p13 was carried out in parallel to loss of heterozygosity (LOH) analyses applying 5 microsatellite markers on these chromosomes. Overall, deletions on chromosome 9p were found in 47 tumors (90%), at chromosome 9q in 36 tumors (69%) and at chromosome 17p in 3 tumors (6%). There was a very high correlation of the results between FISH and LOH analysis. Ten early superficial papillary tumors showed deletion of chromosome 9p without deletion of 9q, suggesting 9p deletions as a very early event in the development of papillary urothelial carcinoma. Although in four patients, all investigated tumors showed identical genetic alterations and one patient showed no genetic alterations at the loci investigated, in five patients, two or more clones with different deletions were found. In four of these patients, the results are compatible with clonal divergence and selection of different cell subpopulations derived from a common progenitor cell. However, in one patient different alleles in two markers at chromosome 9 were deleted, favoring an independent evolution of two recurring tumor cell clones. In summary, we could show that there is considerable genetic heterogeneity in early multifocal and recurring urothelial carcinoma and demonstrated the occurrence of two independent clones in at least one patient as an indicator of possible initial oligoclonality of bladder cancer.     

Gastric emptying and anxiety in chronic duodenal ulcer

The authors have evaluated in a group of patients afflicted with duodenal ulcer whether anxiety might be in some way linked to gastric emptying. The 23 patients studied were subdivided into two groups: "fast" emptiers (n = 12; t 1/2 less than 90 min) and "normal" emptiers (n = 11; t 1/2 greater than 90 min). There were no significant differences between the two groups for the following parameters: sex, age, marital status, educational level, social class, number of cigarettes smoked per day, type of treatment undergone for the duodenal ulcer and maximal acid output. Anxiety levels measured with the Q4 and QII factors of the Cattell's PF 16 were found to be the same in the two groups. The authors conclude that anxiety does not seem to be linked to gastric emptying in chronic duodenal ulcer.     

Marked karyotype abnormalities in two cases of acute myelogenous leukemia

Two patients with acute myelogenous leukemia with severe chromosome abnormalities are described. The cytogenetic analysis shows the following karyotype: patient No. 1: 41,XY, ?1,?2,?4,?5,?13,?15,?17,?17,?18,?22,+5 markers; patient No. 2: 46,XY,?2,?5,?7,?13,+16,?21,?21,+5 markers. In each patient one set of double minute chromosomes was observed.     

Solid tumor cytogenetics. Progress since 1979

Some of the advances in the past decade in the field of solid tumor cytogenetics are described, with particular reference to nonrandom structural chromosome changes. Although it had been known for many years that meningiomas and salivary gland tumors were associated with changes involving particular chromosomes, it has only quite recently become clear, following the application of suitable culture techniques, that other benign tumors such as lipomas and leiomyomas may also be characterized by specific changes, particularly reciprocal translocations. Reciprocal translocations may also be found in malignant soft-tissue tumors such as liposarcomas (involving 12q as in lipomas) and Ewing's sarcoma. In contrast, the common forms of carcinoma present a more variable picture, although certain chromosomes may undergo nonrandom changes of various types, including translocations, which, however, are generally nonreciprocal. Some of these chromosomes may be quite specific (e.g., chromosome 10 in prostatic and #18 in colorectal cancer), while others appear to be common to many or all types of carcinoma, such as chromosomes 1, 3, 11, and 17, and a small metacentric that may be an i(5p). In carcinoma of the bladder, different chromosome changes may characterize subsets of the tumors. In carcinoma of the cervix, however, the commonly involved chromosomes, 1, 3, ?5, 11, and 17, appear in markers in any combination and are thus not mutually exclusive. Although further study of the chromosome changes in carcinomas is essential to an understanding of their relationship to the molecular changes that are associated with malignant transformation, it can be hypothesized that, while some of the changes result in the duplication of particular genes, e.g., on chromosome 1q, a more important role may be to bring about the loss of chromosomal segments containing tumor-suppressor genes. Evidence from molecular studies that has recently been accumulating for the loss of alleles on, for instance, 3p, 11p, and 17p, which could in part be due to gross chromosomal rearrangements, also strongly suggests the importance of genic loss in malignant transformation. In carcinomas, at least, the changes probably involve a number of genes, each change representing one of the several steps necessary for tumorigenesis.     

The incidence, type, and subsequent evolution of 14 variant Ph1 translocations in 180 South African patients with Ph1-positive chronic myeloid leukemia

A Philadelphia (Ph1) chromosome translocation was found in 180 of 198 cases of chronic myeloid leukemia (CML). A standard t(9;22) was present in 166 patients, 83 of whom were black, 79 white, and 4 of "mixed" ancestry; whereas a variant Ph1 translocation was detected in 14 patients (7.8%), 11 of whom were black and only 3 white. There was a higher frequency of a variant Ph1 among black patients compared with whites. The significantly higher frequency of a variant among our patients compared with surveys from elsewhere could be due to differing environmental agents. Simple variants were detected in four patients. Complex variants were found in eight cases; in one of these patients, only chromosomes #9 and #22 were involved, but a complex rearrangement of chromosome #9 had occurred. A "masked" Ph1 translocation was detected in two cases, both of which showed monosomy #22 because the Ph1 chromosome was incorporated or interchanged with chromosome #9. Karyotypic evolution of the Ph1-positive cell line was observed more frequently in the variant group (71.4%) than the standard group (29.5%). This difference was significant (p less than 0.005). There was no difference in the type of clonal changes seen in standard and variant groups. The majority of clonal changes were observed during the acute stage in both groups. In the variant group, there was no obvious correlation between the type of variant, type of clonal change, blast morphology, or survival. Their initial survival pattern resembled that of Ph1-negative cases, but those patients who survived longer than 1 year showed a survival trend similar to standard Ph1-positive cases. Possible explanations for the specificity of chromosome #22 involvement and the constancy of the 22q11 breakpoint in all these variant translocations are discussed.     

Chromosomal aberrations in malignant gastrointestinal stromal tumors: correlation with c-KIT gene mutation.

Gastrointestinal stromal tumors (GISTs) are distinctive, KIT positive mesenchymal neoplasms. The genetic alterations leading to the malignant behavior of these tumors are not well known. In this study, we looked for recurrent numerical chromosomal changes, which may be associated with malignant GISTs, using interphase fluorescence in situ hybridization (FISH). Fourteen malignant primary tumors and two intra-abdominal recurrences were analyzed. Nine benign tumors were studied for comparison. In all cases, the presence of mutations in exons 9, 11 and 13 of the KIT gene were evaluated. Sixteen centromeric enumeration probes (CEP) for chromosomes 1, 2, 3, 4, 6, 7, 8, 9, 10, 11, 12, 15, 16, 17, 18, and X and three locus specific probes (LSI) for 22q11.2 (BCR-locus), 13q14 (RB1-locus) and 14q32 (IgH-locus) were used. The most common changes seen in malignant GISTs were losses of 14q32 and 22q11. However, these changes were commonly detected in benign tumors and represent early changes related to the pathogenesis of GISTs. Losses of chromosomes 1 and 9 were the only recurrent numerical changes seen exclusively in malignant GISTs. Other recurrent numerical changes seen predominantly in malignant tumors were gain of chromosome 8 and losses of chromosomes 7 and 15. The concurrent loss of chromosome 7 and gain of chromosome 8 (in 4 cases) was never seen together with loss of chromosomes 9 or 15 and only once with loss of chromosome 1. Mutations in KIT were found in the majority of malignant GISTs (64%) confirming a previously shown correlation between presence of such mutations and malignancy. KIT mutations were seen in four of five malignant GISTs with loss of chromosome 9, but only in one of four malignant tumors with loss of chromosome 1. These observations may reflect the different pathways leading to malignant transformation of GISTs.     

Chromosome studies in two human brain tumors

The cytogenetic findings based on G- and C-banding in two human brain tumors (a meningioma and an astrocytoma) are reported. Both tumors were characterized by hypodiploid modal numbers (45 and 40 chromosomes, respectively), chromosome 22 abnormalities, and the presence of several markers. This observation supports the hypothesis of the association of No. 22 chromosome abnormalities with tumors of the brain.     

Numerical chromosome changes in 165 malignant tumors. Evidence for a nonrandom distribution of normal chromosomes

The numbers of normal copies of each of the chromosomes in representative karyotypes from 165 malignant tumors of the bladder, breast, cervix, colorectum, and testis studied in this laboratory or described in the literature were assessed to determine whether particular chromosomes were over- or underrepresented. For each chromosome, the mean number of copies was expressed as a percentage of the number expected on the basis of the total number of chromosomes in the karyotypes. The most highly represented autosomes in the tumors as a whole were, in descending order of frequency, numbers 7, 20, 12, 19, 21, and 3, while those most underrepresented were numbers 10, 1, 4, 5, 14, 17, 11, and 18. In tumors of males, the Y tended to be underrepresented. The X was highly represented in the testicular tumors (there were usually two or more copies) and in colorectal tumors of males, but not in the other tumor categories studied. For the tumors as a whole, statistically significant differences could be demonstrated between pairs of autosomes that were at opposite ends of the frequency range. Differences between tumors at the different sites studied were not demonstrable. It is suggested that the determination of the number of normal copies of chromosomes, i.e., whether there are more or fewer than expected, may usefully complement observations on structural changes by reflecting the presence of oncogenes and tumor-suppressor genes, respectively. It may also point to chromosomes that are involved in significant genic changes in which cytogenetic observations on structural changes are equivocal.     

Correlation of chromosome abnormalities with presence or absence of WT1 deletions/mutations in Wilms tumor

Of 40 Wilms tumors with chromosome abnormalities, 6 were hypodiploid, 10 were pseudodiploid, 7 were hyperdiploid with 47 to 49 chromosomes, and 17 were hyperdiploid with 50 or more chromosomes, mostly including +12. WT1 deletions/mutations were found in one hypodiploid, eight pseudodiploid, and one hyperdiploid (47-49 chromosomes) tumor, but in none of the hyperdiploid (> or =50 chromosomes) tumors. Of the 10 tumors with WT1 abnormalities, 6 had a homozygous WT1 deletion, 1 had a nonsense WT1 mutation and loss of heterozygosity at 11p, 1 had an intragenic hemizygous WT1 deletion without detectable WT1 mutation, and 2, which occurred in Wilms tumor-aniridia-genitourinary abnormalities-mental retardation syndrome patients, had a hemizygous deletion and a missense or frameshift mutation of WT1. Six of the nine tumors with homozygous or hemizygous WT1 deletions had chromosome aberrations involving chromosome band 11p13 in one of the two chromosomes 11. While one hypodiploid and one pseudodiploid patient died of the disease, and one hyperdiploid (47-49 chromosomes) patient was alive in nonremission, all hyperdiploid (> or =50 chromosomes) patients had no evidence of disease at the last follow-up. Our data show that chromosome aberrations are closely correlated to WT1 abnormalities and suggest that hyperdiploid (> or =50 chromosomes) Wilms tumors may be characterized by the absence of WT1 abnormalities and possibly also by a favorable prognosis.