Cytogenetic diagnosis of cancer: abnormalities of chromosomes and polyploid levels in the bone marrow of patients with small cell anaplastic carcinoma of the lung.

The chromosomes of metastatic cells and polyploid levels in the bone marrow of 26 patients with small cell anaplastic carcinoma were studied by direct bone marrow preparation and trypsin-Giemsa banding. Eighteen of these patients had received no tumor therapy and 8 had had chemotherapy and/or radiation therapy; 18 patients, including 5 who had received therapy, had karyotypic abnormalities with or without elevation of the polyploid level. Modal numbers and chromosome abnormalities were highly variable in treated and untreated patients. Modes ranged from hypodiploid to polyploid, but polyploid modes were the most frequently observed abnormal modes. Polyploid modes were not seen, however, in post-therapy patients with the exception of one who had received radiation therapy to the mediastinum for only 4 days prior to withdrawal of the specimen for chromosome analysis. Ten patients had elevated polyploid levels that ranged from 4.24 to 44.8% and always occurred in conjunction with karyotypic abnormalities. Both aneusomy (abnormal number) of normal chromosomes and structural aberrations (markers) occurred frequently. Some markers were consistent within an individual, but other variable aberrations were also typically present. Very few markers were common to 2 or more patients. The no. 1 chromosome participated in marker formation in 14 of the 18 patients with karyotypic abnormalities. Of the 26 patients, 5 were negative for metastasis to the marrow by pathologic examination but positive by cytogenetic diagnosis, whereas none were positive by pathologic examination and negative by cytogenetic diagnosis; this demonstrated that cytogenetics may be used as a rapid adjunct diagnostic procedure for the detection of metastasis in the marrow.

Karyotypic evolution associated with loss of tumorigenicity.

A reproducible association between loss of tumorigenicity and specific karyotypic changes was described in cell culture lines SLU-5 and DMS-402 established from mouse plasmacytoma MOPC-21 carried in BALB/c mice. The defect in chromosome no. 15, which has been specifically associated with mouse myelomas, was neither corrected nor eliminated in the karyotypic evolution that occurred simultaneously and progressively with the grandual loss of oncogenicity.

Karyotype, marker formation, and oncogenicity in mouse plasmacytomas.

Two common chromosome markers in the 2 plasmacytomas previously examined by Giemsa banding were consistently present in the mouse plasmacytoma X-5563, a transplantable hypertetraploid tumor of spontaneous origin in C3H mice. The 2 markers were found in both induced and spontaneous tumors and in either BALB/c or C3H mice. The derived cell line had 17 fewer chromosomes than the X-5563 tumor and was oncogenic, and its modal karyotype was identical to that of the tumor transmitted by the inoculation of the cell line. The homogeneity of a slight karyotypic modification in a second tumor suggested a possible clonal origin of that tumor. The high frequency of centric fusions between homologues and the structure of certain markers suggests that homologue association may precede marker formation. We proposed a second mechanism of marker formation, selective regional elongation, to account for the larger number of markers with proximal or distal elongations without evidence of translocation and for the observed alterations in length and banding pattern of markers after growth in vitro. Comparison of MOPC-21, MOPC-315, and X-5563 tumors showed preferential involvement of certain chromosomes in marker formation, an inferred association of the 2 common markers with an early stage in the origin of the 3 plasmacytomas, and consistent loss of an X chromosome. Loss of oncogenicity in cell lines was associated with a number of karyotypic changes, but did not require the loss of the characteristic markers or additional copies of a specific normal chromosome.

A specific chromosome breakpoint associated with mouse plasmacytomas.

The chromosomes of uncultured cells of the near-diploid mouse plasmacytoma MOPC-31C were studied. The modal number of chromosomes was 44. The tumor lacked two marker chromosomes, reciprocal translocation [rcp t(12; 15)], that in previous studies were found to be common to 3 other uncultured myelomas and 1 cultured mouse myeloma. Through the formation of two markers, rcp t(6; 15), unique to this tumor, however, the tumor shared with other tumors and their specific markers a common breakpoint in chromosome "15 at band D3/E. This breakpoint has been found in all mouse plasmacytomas examined with banding thus far and is considered of possible importance in the development of this tumor.

Isolation and characterization of a hormone-producing cell line from human small cell anaplastic carcinoma of the lung.

A continuous cell culture line was established from a bone marrow metastasis of small cell anaplastic carcinoma of the lung. The cultures were characterized by light and electron microscopy, and an unusual concentric arrangement of cells was observed, both in sectioned material from the patient's tumor and from the cell cultures. The cells had two types of specialized cell junctions and contained secretory-like granules of the type described in neuroendocrine cells. Lactic dehydrogenase isozyme patterns were the same as those observed in normal human serum, and the karyotype revealed the presence of several marker chromosomes. Vasopressin was present in the cells and secreted into the culture medium in the absence of neurophysin, as shown by the immunoperoxidase technique and radioimmunoassay. Oxytocin was also absent from cells.

A cytogenetic study of familial medullary carcinoma of the thyroid.

Familial medullary carcinoma of the thyroid (MCT) is a malignant neoplasm of the calcitonin-producing C-cells of the thyroid gland, inherited in an autosomal dominant pattern with a high degree of penetrance. We have studied 26 individuals from 5 New England families with MCT with the goal of finding a cytogenetic test useful in the detection of the presence of the gene. G-banded metaphase spreads, segregational errors of chromosomes, breakage in untreated cells, breakage in bleomycin-challenged cells, and high-resolution banding of prophase/prometaphase spreads were examined; one or more of each of these procedures were studied in at least one affected individual in each of 5 families. All specimens were coded, and slides were scored blindly. All tests were negative with respect to significant differences from controls. We did not observe the minute deletion from the short arm of chromosome 20 that was reported by other investigators to occur in individuals with multiple endocrine neoplasia types 2A and 2B which include MCT. Contrasting results from our study and several other cytogenetic studies of MCT may reflect genetic heterogeneity among the families studied and/or differing environmental factors in various geographic locations that are possibly associated with initiation and onset of the disease.

Non-reciprocal translocation (5;15), isodicentric (15) and Prader-Willi syndrome.

A non-reciprocal translocation (5;15) and an isodicentric (15) resulting in trisomy 15pter----15q1?3 and monosomy 5qter [46,XY,-5,-15,+der(5)t(5;15) (5pter----5q35::15q13----15qter),+idic(15) (pter----q1?3::q1?3----pter)] was found in a 28-year-old profoundly retarded male resident of a state institution. Early developmental history and childhood and adult physical findings resembled those of Prader-Willi syndrome (PWS) patients. The parents' unbanded chromosomes were normal. Blood groups of parents and propositus were uninformative with regard to identifying gene deletions or duplications.

Mixoploidy in humans: two surviving cases of diploid-tetraploid mixoploidy and comparison with diploid-triploid mixoploidy.

We report on 2 cases of diploid/tetraploid (2n/4n) mixoploidy in surviving females, 11 and 21 years old. Both individuals manifested severe mental retardation, reduced peripheral limb muscle bulk, asymmetric growth deficiency, seizure disorder, and skin pigmentary dysplasia. Previous lymphocyte karyotypes had been normal on 2 occasions, but when skin fibroblast karyotypes were done, 33% of the cells were tetraploid on the younger woman, and over 60% were tetraploid in the older woman (on 2 separate occasions). In both individuals, the distal limbs and digits were long and thin, with reduced small muscle bulk. The similarity in distal limb findings prompted reexamination of the younger woman's chromosomal constitution in skin fibroblasts. We concluded that the clinical findings in these cases are unique and similar, and we caution clinicians about uniformly dismissing tetraploidy as artifactual in amniocytes from normal patients, especially since this phenotype would be very difficult to detect, even with directed prenatal ultrasonography. We compare the 2n/4n phenotype with that in diploid/triploid (2n/3n) mixoploidy and note subtle differences which might be detected postnatally. These findings should be useful in guiding clinicians on when to request skin fibroblast karyotypes in mentally-deficient individuals with asymmetric growth deficiency and pigmentary skin variation.

Ring chromosome 12.

A ring chromosome 12 (p13.3q24.3) was observed in all cells analyzed from skin fibroblasts and the peripheral blood of a 19-year-old man initially referred for developmental delay with expressive language deficiency. Other phenotypic anomalies included growth deficiency, multiple café-au-lait spots, mild pectus excavatum, glandular hypospadias, left esotropia, clinodactyly of the fifth fingers, and hypothyroidism with elevated antithyroid antibodies. The four previously reported cases of r(12) support the theory of a general ring phenotype which is manifested independently of the specific autosome involved and which is characterized by growth failure, moderate mental retardation, and lack of other major phenotypic anomalies. Breakpoints in all cases of r(12) have been assigned to the telomeric regions, suggesting minimal deletion of chromosome material.

Familial translocation 5;14 resulting in an unbalanced offspring.

We report on an infant with multiple congenital anomalies possessing a derivative 14 chromosome in excess of the normal complement, resulting from transmission of a familial t(5;14)(p13;q22). The proposita's phenotypically normal mother, mentally retarded half-brother, and fetal sib are carriers of the apparently balanced translocation. Previous cases of similar familial t(5;14) are reviewed. The proposita's phenotype is characterized by failure to thrive, developmental retardation, cleft palate, congenital heart anomaly, abnormal hands and feet, unusual face with abnormal ears, and recurrent respiratory infections. The proposita died at age 9 months and postmortem examination showed multiple central nervous system, cardiopulmonary, gastrointestinal, and genital malformations. Our proposita's phenotype is attributable to contributions from both chromosomes and is consistent with the consequences of both the dup(5p) and dup(14q).

Familial t(11;13)(q21;q14) and the duplication 11q, 13q phenotype.

Cases of duplication of distal 11q or proximal 13q have been reported independently. A specific translocation resulting in duplication of distal 11q, [der(22)t(11;22)(q23;q11)], has been documented in over 40 cases. We report on a male fetus with chromosomal excess of both distal 11q and proximal 13q resulting from a familial translocation. This case supports the causal association of duplication 11q with neural tube defects.

Isodicentric chromosome 18q as sole abnormality in a histologically normal bone marrow from a patient with diffuse large-cell non-Hodgkin's lymphoma.

Cytogenetic analysis was performed on the histologically and immunophenotypically normal bone marrow (BM) of a 33-year-old woman with non-Hodgkin's lymphoma (NHL) before BM harvest. Unstimulated 24- and 48-hour cultures produced only normal metaphases. A pokeweed mitogen (PWM)-stimulated 48-hour culture, however, showed a clonal isodicentric chromosome 18q as the sole abnormality, suggesting a role for this approach in detection of submicroscopic BM involvement by B-cell NHL.

Cytogenetics of medullary carcinoma of the thyroid.

Medullary carcinoma of the thyroid (MCT) is a dominantly inheritable neoplasm derived from intrathyroid C cells. The cytogenetics of this tumor has been only sparsely and indirectly studied previously. This article describes the chromosomes of primary MCT tumor tissue cultured with colcemid for 48 hr, metastatic tumor in lymph node cultured for 7 days from the same patient, and of primary tumor tissue cultured for 3 weeks from a second patient. The modal numbers were 42-44 in all 3 specimens. Karyotypes from the metastastic tumor were similar to those of the primary tumor. Karyotypes of the two primary tumors, however, differed from each other, having in common only the modal numbers and the loss or structural alteration of a #22.

Hypodiploid, pseudodiploid, and normal karyotypes prevail in cytogenetic studies of medullary carcinomas of the thyroid and metastatic tissues.

Medullary carcinoma of the thyroid (MCT), often a dominantly inherited neoplasm, derived from intrathyroid C-cells of neural crest origin, is one of the solid tumors least studied cytogenetically. The cells are difficult to grow in culture, only two cell lines having ever been established. Cytogenetic studies of only 5 tumors have been reported previously. In this paper we report on the cytogenetic analyses of 8 specimens of primary and/or metastatic MCT tumor tissue from 6 patients with familial disease, including more recent metastatic tumors in lymph node and femur of a patient whose thyroid and earlier lymph node metastases were described previously. Some of these specimens were harvested sequentially over time. Hypodiploid or diploid modal numbers prevailed with normal, pseudodiploid, or hypodiploid karyotypes.

Fragile sites and high-resolution chromosome studies in multiple endocrine neoplasia type 2A.

Affected individuals from four kindreds with multiple endocrine neoplasia type 2A syndrome (MEN-2A), were studied for the possible existence of a specific fragile site that might be associated with the MEN-2A gene. The chromosomes were also examined with high-resolution banding with particular emphasis on those chromosomes (#1, 10, 20, and 22) that have been implicated by previous studies from several laboratories as being associated with this disease. There was no evidence for a unique fragile site or a unique high-resolution banding pattern in subjects with MEN-2A. These findings, in combination with all previous cytogenetic studies, indicate that it is unlikely that current techniques will be useful in developing a simple cytogenetic test for this disease.

Cytogenetics of small cell carcinoma of the lung.

Nineteen cell lines derived from various malignant tissues of 15 patients with small cell carcinoma of the lung (SCCL) have been studied. The results showed heterogeneity in all cell lines, with no one consistent abnormality among them. Cell lines from 11 of the patients had minute and double minute chromosomes, and cell lines from 2 patients had abnormally banding regions, designated as ABRs, as distinguished from homogeneously staining regions (HSRs). The latter 2 and several of the former cell lines were derived from specimens taken before the patients were placed on therapy. All but 2 of the cell lines had a constant marker load, consisting of 24%-35% of the complement. Some markers remained stable through months and years of culture life, while other markers came and went. Chromosomes #1, #6 and #11 were most frequently involved in marker formation in the cell lines, and these were compared to similar markers in direct bone marrow preparations. Chromosome #1 markers were of variable structure, whereas #6 and #11 most often took the form of 6q- and 11p+ markers, with breakpoints most frequently at 6q23-25 and 11p11-12. A 3p- marker was found in a minority of cell lines. All of these markers were also found in direct marrow preparations from some patients with SCCL. Nonmonoclonal tumors arose from inoculation of bimodal cell lines into nude mice, but population selection by undetermined mechanism was evident. Cytogenetic parameters showed no positive correlation with hormone production by these cell lines.

Chromosome identification of the rusty-spotted cat (Felis rubiginosa): one more down and four to go.

The chromosomes of the rusty-spotted cat, Felis rubiginosa, of Sri Lanka (Ceylon) were analyzed. Although the 38-chromosome complement of F rubiginosa closely resembles that of several other species, it seems to be unique with only minor variations in the D and F chromosomes. This reduces, from 5 to 4, the number of the 37 recognized world feline species not yet cytogenetically studied. Despite great differences in size and distribution, the feline species of the world are one of the most homogenous families of mammals, with only subtle chromosomal differences separating most species. By comparative cytogenetic analyses, it is becoming possible to map the probable evolutionary pathway of the world's cats.

Chromosome identification of the flat-headed cat, Felis planiceps: one more down and five to go.

There are 38 recognized feline species in the world. With successful chromosome analysis of the flat-headed cat, Felis planiceps or Prionailurus planiceps, from the secluded river banks and lower mountain slopes of the jungles of Malasia, Borneo, and Sumatra, there remain only 5 rare and endangered species yet unstudied. Although some, including the flat-headed cat, have somewhat distinctive chromosome configurations, all cats thus far karyotyped show 38-chromosome counts, excepting 5 new world "tropicals" which have 36. A pattern of probable feline mutations can be read from the chromosome variations in the different species.