Cytogenetic analysis in human breast carcinoma. I. Nine cases in the diploid range investigated using direct preparations

Cells in mitosis were found in 51 of 110 (47%) breast tumor samples; karyotypes of nine tumors in the diploid range are presented. The simplest stemline karyotype found was 46,XX,?16,+del(1)(qter→p21). The chromosome homologues most frequently lost were #8, #13, and #16. Monosomy or partial monosomy for chromosome #16 was seen in six cases, including the two simplest and chromosome #16 might be of relevance for initiation of malignant transformation in breast carcinoma.The only chromosome feature common to all nine breast carcinomas was the presence of a marker involving the long arm of chromosome #1, the region shared by all being 1qter→1q21.     

人脑垂体瘤染色体分析

为探讨垂体瘤及癌的发病机理与肿瘤演进过程中的细胞遗传学特征，对１５例人脑垂体瘤进行了染色体分析。实验结果表明染色体数目的分布从２４～２１０，２０％病例干系细胞为二倍体染色体，而８０％病例干系细胞为非二倍体染色体，其中亚二倍及超二倍体以上染色体病例分别为１３％和６７％。１０例人脑垂体瘤Ｇ显带核型分析，９０％病例具有２２号染色体减少及７号染色体增加，侵袭性垂体瘤７号染色体增加明显。６０％病例具有８、９号染色体的增加。提示人脑垂体瘤在病理形态学虽属良性肿瘤，而细胞遗传学却呈恶性表现。作者认为２２号染色体的丢失，７、８和９号染色体的增加，可使调控细胞增殖，分化基因缺失，癌基因活化，可能与该肿瘤的发病机理有关，７号染色体的明显增加可能为肿瘤演进中的表现。     

Chromosomal doubling: the significance of polyploidization in the development of human tumors: possibly relevant findings on a lymphoma

Recent molecular evidence points to defects in cell cycle checkpoints as one of the most important events in the transformation of normal to malignant cells. A byproduct of, if not a critical step brought about by, these defects is the occurrence of polyploidization; near-tetraploid and near-octoploid cells are a common feature of cancers, and the neoplastic stemline may itself attain a high (e.g., near-tetraploid) value. This short review cites cases in which polyploidization is frequent, even at an early stage of tumor development, and considers the probability that, once a high stemline has arisen, there is increased instability with the likelihood of further chromosome changes. A possible example of the latter is a lymphoma in which tetraploid and hypotetraploid metaphases were found, the latter, interestingly, showing an apparently preferential loss from tetraploidy of chromosome 10. It appears, therefore, that a stemline was emerging consequent to (a) chromosome doubling resulting in tetraploid cells, and (b) the appearance of a hypotetraploid line in which chromosome 10 was under-represented. Alternately, there might have been a repeated loss of chromosomes from tetraploid cells that preferentially included chromosomes 10.     

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.     

Noninvolvement of chromosome 16 in karyotype evolution of acute myeloid leukemia in a patient with a heritable fragile site on 16q22

A fragile site on the long arm of chromosome #16 (q22) was detected in a 24-year-old man with pancytopenia. During the course of the disease he developed an inverted duplication of region q11-12 of chromosome #1 and a translocation between chromosomes #9 and #13: t(9;13)(p22;q32). These abnormalities, as well as an additional iso-like marker chromosome that consisted of one normal 9p and the abnormal 9p arm, were detected in Epstein-Barr nuclear antigen-positive B-cell cultures. Two years later, evolution of the abnormal clone with loss of chromosome #7 and, subsequently, chromosome #22 occurred in connection with development of acute myeloid leukemia. Although the heritable fragile site on chromosome #16 was present in all cell populations investigated, it was not involved in the evolution of the abnormal karyotype. This fragile chromosome #16 also was found in 4 of 11 family members in whom chromosome analysis was performed, thus suggesting this aberration was inherited in a dominant autosomal pattern. The incidence of the heritable fragile site in normal and leukemic cells of the patient, as well as stimulated blood cultures of his relatives, are reported. In addition, the possible relationship between this constitutional chromosome breakage syndrome and the occurrence of leukemia is analyzed.     

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.     

A der(14)t(1;14)(q12;p11) in chronic myelomonocytic leukemia

Duplication of the long arm of chromosome 1 (1q) is widely reported in human neoplasia, including the myelodysplastic syndromes (MDS). So far, it has not been described as a single aberration in the chronic myelomonocytic leukemia (CMML), a subtype of MDS. Rather, trisomy 1q was always a part of complex chromosome changes affecting the subtypes of MDS other than CMML. We report on a patient with CMML with an unbalanced translocation of the entire 1q onto the short arm of chromosome 14 as a sole cytogenetic abnormality. Fluorescence in situ hybridization (FISH) analysis with an alpha-satellite probe for the paracentric region of the long arm of chromosome 1 confirmed the presence of trisomy 1q in a derivative chromosome, der(14)t(1;14)(q12;p11). The discrepant results between the metaphase cytogenetics (100% abnormal) and interphase cytogenetic (71% nuclei with 3 signals) suggest that trisomy 1q, even in the absence of additional cytogenetic changes, has a sufficient leukemogenic potential to confer a proliferative advantage on hematopoietic cells committed to monocyte stemline both in vitro and in vivo. The literature data on partial and complete trisomy 1q in CMML is reviewed.     

Chromosomal patterns in Warthin's tumor. A second type of human benign salivary gland neoplasm

The cytogenetical observations in eight successfully cultured human adenolymphomas are reported. When the results were considered with those of two previously reported cases, three main stemline groups could be distinguished: (a) one with a normal karyotype and noted as a primary or secondary stemline in all hitherto studied tumors; (b) a second group with only numerical changes, either loss of the Y chromosome or trisomy or monosomy 5; and (c) a third group with only structural changes, as a rule with one or two reciprocal translocations. With regard to the last group, studies of many more cases are necessary to decide whether distinctive subgroups exist. Analyses using molecular methods are also urgently needed to clarify whether the normal stemline cells contain submicroscopic changes.     

Cytogenetic analysis of a plexiform fibrohistiocytic tumor

A plexiform fibrohistiocytic tumor was analyzed cytogenetically after short-term in vitro culture. The stemline karyotype of this tumor was interpreted as 46,XY, -6, -8,del(4)(q25q31), del(20)(q11.2), + der(8)t(8;?)(p22;?), + mar. We believe this to be the first report of chromosome findings in this recently described histological entity.     

Karyotypic differentiation in a spontaneous mouse B-cell leukemia

The karyotype of a spontaneous mouse B-cell leukemia passaged intravenously several times in succession into syngeneic recipients was determined. Peripheral lymphocytes from a control animal and from animals in passage 1 and 7 were stimulated by the mitogen lipopoly-saccharide, and studied by Giemsa banding methods. In passage 1, the cultured lymphocytes consisted of 4 clones, one with a normal karyotype (1 cell out of 32), and three with multiple chromosome changes. One clone, representing about 10% of all mitoses, had 39 chromosomes with monosomy for chromosomes 12 and 13, two reciprocal translocations, and a small marker chromosome. One of the translocations had occurred between chromosomes 4 and 15, the other between an X chromosome, and one chromosome No. 8. Another clone had 37 chromosomes (6% of all mitoses), and the remaining clones had 36 chromosomes (81% of all mitoses). These clones had the same chromosome changes as the clone with 39 chromosomes, three additional monosomies (6, 8, and 15), and three new translocations. In passage 7 the clone with 36 chromosomes represented about 30% of the cultured cells, while 70% of the cells had 35 chromosomes; the reduction in number was caused by a 17;19 translocation. The cultures from the control animals had a normal karyotype.     

Chromosome banding analysis of a new stemline of hypodiploid Hirosaki sarcoma established by single cell transplantation

A new hypodiploid stemline of the long-lasting ascites sarcoma, Hirosaki sarcoma (HS), was established by means of single-cell transplantation, and its G-, C- and NOR-banding patterns were compared with those of the predecessor stemline, with special notice taken of the structural alteration of marker chromosomes. Their chromosomal relationship was well explained by the rearrangement, t(M7pM8q): Translocation of the short arm of marker M7 to the NOR of marker M8 produced a new marker M8t, and the translocation-carrying variant cells grew into a subline. A new translocation-carrying stemline was formed from this subline by single-cell transplantation. Judging from the size and number of silver-positive NORs, the NOR activity was significantly higher in this stemline than in its predecessor. The increased NOR activity might have been partly responsible for the growth of a translocation-carrying variant cell into a new subline. Single-cell-mass transplantation may be one of the most effective systems for studying the proliferative ability of subline cells and for understanding the mechanism of stemline formation in transplantable rat ascites sarcomas.     

A serially transplantable human giant cell glioblastoma that maintains a near-haploid stem line

We have karyotyped a human giant cell glioblastoma removed from an 11-year-old girl and have established from it a subcutaneously transplantable line in athymic nude mice. The original tumor contained near-haploid cells with 25 or 26 chromosomes, including two copies of #1, (7 or 7p+) and #18. There were also hyperdiploid (49-52) cells that were tetraploid for these same three chromosome types; doubled versions of the hyperdiploid population were also seen. The stemline of the mouse-grown tumor was 26,X, +1, +7p+, +18 in the first passage and has remained consistently near-haploid through ten serial in vivo passages. Growth stabilization has occurred with an average latency of less than 3 months. This transplantable line is available for evaluating chemotherapeutic responsiveness of human giant cell glioblastoma and for studying near-haploidy in solid human tumors.     

Burkitt cell acute lymphoblastic leukemia with partial expression of T-cell markers and subclonal chromosome abnormalities in a man with acquired immunodeficiency syndrome

A 45-year-old white male, bisexual, with a 2-year history of acquired immunodeficiency syndrome (AIDS) prodrome, developed a Burkitt cell-like acute lymphoblastic leukemia (ALL). Marker studies of marrow blasts show an unusual and possibly unique pattern, in that an unequivocal monoclonal B cell leukemia, having K-IgM with HLA-DR and B cell subset antigen (BA-1) expression, was superimposed with a mature suppressor T cell marker profile (pan-T, mature T, and suppressor/cytotoxic T antigens). The leukemic blasts were totally negative for terminal deoxynucleotidyl transferase (TdT), human T cell leukemia-lymphoma virus (HTLV) p19 antigen, and other immunoglobulin isotypes. Chromosome analysis of marrow cells disclosed that 70% of the cells had 47,XY, + 12,t(8;14)(q24;q32) chromosome complement, and 30% of the cells had a 47,XY, + 12,dup1q + (q22-31),t(8;14)(q24;q32). The consistent finding of the specific chromosome rearrangement (8/14 translocation) in all abnormal cells suggests that the cells were derived from a common precursor. With regard to the partial T cell marker expression, the significance of these markers in B cell leukemia is unclear, as is their relation to the additional chromosome abnormalities that apparently developed in the process of clonal evolution.     

Cytogenetic characterization of a colon adenocarcinoma from a familial polyposis coli patient

We report on the cytogenetic findings in a fresh biopsy of a colorectal adenocarcinoma in a patient with familial polyposis coli (FAP). The stemline had 45 chromosomes with several clonal chromosome aberrations including a der(17)t(17;?), and -18, which are the two most recurrent aberrations in sporadic colorectal carcinomas. This finding is discussed in relation to the chromosomal changes described in sporadic colorectal carcinomas and in FAP cell lines.     

Flow cytometric analysis of stemline heterogeneity

The stemline heterogeneity of malignant tumors is closely connected with tumor aneuploidy. Both features can be characterized by either chromosome analysis or DNA cytophotometry. DNA analysis may be performed either by single cell photometry or by flow cytometry, of which the respective advantages and drawbacks are presented. Preliminary results of flow cytometric DNA analysis in malignant neoplasms are discussed; the possibilities of multiparametric measurement for quantitative analysis of various biochemical and antigenic properties in DNA stemlines are considered.     

Consistent cytogenetic aberrations in hepatoblastoma: A common pathway of genetic alterations in embryonal liver and skeletal muscle malignancies?

Cytogenetic analyses of four consecutive hepatoblastomas revealed near-diploid stemline karyotypes with relatively simple chromosome aberrations. Cytogenetic abnormalities shared by each tumor included trisomy for all of part of chromosome 2 and trisomy for chromosome 20. In two cases, partial trisomy for chromosome 2 resulted from direct duplication of long arm material with the shortest region of overlap being 2q23-2q35. In one tumor, each metaphase also contained a variable number of double minute chromosomes found not to derive from NMYC amplification. Interestingly, trisomy for 2q and trisomy 20 are also characteristic events in pediatric embryonal rhabdomyosarcomas. Furthermore, others have reported loss of heterozygosity for the short arm of chromosome 11 in both hepatoblastoma and childhood embryonal rhabdomyosarcoma, and both these malignant diseases are associated with Beckwith-Wiedemann syndrome. These cytogenetic and molecular findings suggest a parallel pathway of genetic steps in the initiation and/or progression of tumors of embryonal liver and skeletal muscle.     

Noninvolvement of a constitutional heritable fragile site at 10q24.2 in rearranged chromosomes from rectal carcinoma cells

A fragile site in chromosome band 10q24.2 was found in the lymphocytes of a patient ascertained for rectal carcinoma. The karyotype of 110 R-banded tumor cells was performed, showing two stemline formulas: 46,XXY,-1,-18,+20,der(6),t(1;6)(q21.1;q22.3),i(17q) and 46,XY,-1,-18,+8,+20,der(6),t(1;6),del(2)(p1600p22),i(17q). These findings are in agreement with our previous studies, which reported that the rearrangement of chromosome #17 and the loss of chromosome #18 are recurrent anomalies in colorectal carcinomas. In addition to these rearrangements, other anomalies were occasionally observed in tumor cells, but no breakages nor rearrangements involving band 10q24.2. The relationships between fragile sites and cancer breakpoints are discussed.     

Ph-positive CML in a family with a constitutional Robertsonian translocation 14;15

A patient with CML in chronic phase was admitted to our center for bone marrow transplantation. Cytogenetic analysis of bone marrow cells revealed a Ph chromosome due to a standard t(9;22) and a Robertsonian t(14;15). This Robertsonian translocation was also found in PHA-stimulated lymphocytes of the patient's sister, the donor of the bone marrow. A chromosome study of the whole family proved the constitutional character of the t(14;15) abnormality and showed that it was inherited from the father's side. All family members were phenotypically normal with normal mental status. Female carriers of the t(14;15), as well as wives of male carriers, had a high incidence of miscarriages and early death of their offspring. The occurrence of Ph-positive CML in a patient with a Robertsonian t(14;15) might indicate increased risk for the development of leukemia in patients with this constitutional chromosome abnormality. This assumption however, is limited by the rarity of the Robertsonian translocations.     

Chromosomal composition of four permanent cultured cell lines derived from human gliomas

Four permanent cell lines derived from malignant human gliomas were karyotyped using Giemsa-trypsin banding. D-65 MG had a stemline with 44 chromosomes, including 11 markers: 1p+, 2q-, 3p-, 3q+, 4p-, 9q-, 11q+, 15q-, 17p+, 21p+, 22q-. The net effect after accounting for fragments in markers was: +8, -10, -16 -X. D-32 MG had chromosome counts 90-91 without a distinct stem karyotype. Modal cells contained from 3 to 5 copies of the normal autosomes and 5 markers: 1q-, 3q-, 7q-, 13q-, 18q-. D-32 MGCl2 had a complex karyotype containing 78-82 chromosomes. There was no stemline, and modal cells varied from one another primarily in their set of marker chromosomes. A total of 23 markers were seen in this line, 17 of which were present in most modal cells. They were partially characterized as: 1q+, 1q+, 2q-, 5q+, 7p-, 7q-, 8p+, 8p+, 9p+, 12p+, 14q-, 16q+, 19q+, 19q+, a small submetacentric chromosome of undetermined origin and two small isochromosomes, i(Dp or Gp) and i(17p or 18p). A-172 MG had a modal peak of 77 chromosomes within which no two cells were exactly alike. Ten markers seen in modal cells were: 1p-, 4p+, 6p+, 6p+, 6q-, 7p-, 9p-q+, 13q+ 14p+, 22q+. There were no normal copies of chromosomes #1, #6, #9, #14. These four glioma-derived cell lines possess unique karyotypes, but each displays some combination of the numerical and structural deviations generally associated with established glioma lines.     

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.