GLI3 encodes a 190-kilodalton protein with multiple regions of GLI similarity.

The GLI oncogene, discovered by virtue of its amplification in human tumors, encodes a sequence-specific DNA-binding protein containing five zinc fingers. We have now characterized one member of a family of GLI-related zinc finger genes. A previously identified fragment of GLI3 genomic DNA was used to localize GLI3 to chromosome 7p13 and to isolate cDNA clones. Sequence analysis of these clones and identification of the GLI3 protein by using polyclonal antisera demonstrated that GLI3 encodes a protein of 1,596 amino acids and an apparent molecular mass of 190 kilodaltons. Amino acid sequence comparison with GLI demonstrated seven regions of similarity (53 to 88% identity), with the zinc fingers representing the most similar region. Furthermore, when produced in vitro, the GLI3 protein bound specifically to genomic DNA fragments containing GLI-binding sites. Amino acid sequence comparison with the product of another member of the GLI family, the Drosophila segment polarity gene cubitus interruptus Dominant, revealed additional similarity that was not shared with GLI. These studies suggest that the GLI-related genes encode a family of DNA-binding proteins with related target sequence specificities. In addition, sequence similarity aside from the zinc finger region suggests that other aspects of function are shared among the members of this gene family.

The INT1 oncogene is not rearranged or amplified in lipomas with structural chromosomal abnormalities of 12q13-15.

DNA from nine different solitary subcutaneous lipomas, all with clonal abnormalities affecting the chromosome segment 12q13-15, was examined for rearrangement or amplification of a human INT1 gene sequence. INT1 is a putative oncogene that has been localized to the chromosome band 12q13. No rearrangement or amplification could be detected.

No amplification or rearrangement of INT1, GLI, or COL2A1 in uterine leiomyomas with t(12;14)(q14-15;q23-24).

We have studied three uterine leiomyoma tumors, all previously cytogenetically analyzed and shown to have the clonal abnormality t(12:14)(q14-15;q23-24), with the purpose of detecting amplification or rearrangement of three genes that are localized close to the 12q breakpoint region. The genes studied were the two putative oncogenes INT1 and GLI, and the collagen type II alpha 1 gene, COL2A1. No rearrangement or amplification could be detected for any of the three gene sequences.

Rings, dicentrics, and telomeric association in histiocytomas.

We report clonal karyotypic abnormalities in six of 12 cytogenetically investigated malignant fibrous histiocytomas. Four of eight tumors of the pleomorphic subtype had complex clonal chromosome aberrations, including ring chromosomes, dicentric chromosomes, and/or telomeric associations. No common characteristic aberration could be distinguished. Two of four myxoid tumors had clonal changes: One had one to two ring chromosomes and an extra chromosome #2; another had a supernumerary ring chromosome as the sole abnormality.

Multiple karyotypic rearrangements, including t(X;18)(p11;q11), in a fibrosarcoma.

The tumor stem line of a soft tissue fibrosarcoma, histologic malignancy grade III, had 43 chromosomes with several clonal chromosome aberrations, including the three balanced translocations t(X;18)(p11;q11), t(2;15)(p23;q26), and t(7;22)(q11;q13), two partly identifiable marker chromosomes, der(3)dic(3;?)(p11;?) and der(5), one small marker of unknown origin, and loss of one chromosome #11, #18, #19, and #21.

Unbalanced chromosomal rearrangements in a metastasizing salivary gland tumor with benign histology.

Benign metastasizing pleomorphic adenoma (BMPA) is a rare tumor of the salivary glands. Despite benign histopathologic features, it can metastasize and is sometimes lethal. No chromosomal data have been reported for this tumor type. We have by chromosome banding and fluorescence in situ hybridization analysis examined the short-term cultures of three skeletal metastases from a BMPA and identified two related hypodiploid clones: 44,XX,dic(3;22)(p11;q13) or der(3)t(3;22)(p11;q?) add(22)(q?),der(9;21)(q10;q10),der(13)t(1;13)(q11;p13)/45,XX,-3,der(9;21 ) (q10;q10),der(13)t(1;13)(q11; p13),?der(22)t(3;22)(q22;q13), +mar. The karyotypic features of this BMPA thus differ from the characteristic cytogenetic findings in pleomorphic adenomas and carcinomas ex pleomorphic adenoma.

Genomic amplification of CCND2 is rare in non-Hodgkin lymphomas.

Cyclin D2, encoded by CCND2 at 12p13, takes part in the regulation of the cell cycle and has been suggested as a candidate for gene amplification in lymphoid malignancies. CCND2 is often overexpressed in chronic B-cell disorders, and we recently detected genomic amplification of the chromosomal region containing CCND2 in two of three investigated non-Hodgkin lymphomas (NHLs) with cytogenetic abnormalities involving 12p. In the present study, 58 NHLs without karyotypic evidence of 12p aberrations were analyzed by fluorescence in situ hybridization with probes for CCND2. No genomic amplification was found, strongly suggesting that this abnormality is rare in such NHLs.

Multiple karyotypic abnormalities, including structural rearrangements of 11p, in cell lines from malignant melanomas.

Cell lines were obtained from three malignant melanoma patients by culturing cell suspensions from tumor biopsies. A total of six lines (I to VI) were established. One line each was established from the first two cases. Lines III and IV were established from two different methyl cellulose colonies derived from the primary tumor of case 3; line III was from a non-pigmented and line IV from a pigmented colony. Cloning of line IV resulted in two highly malignant (IV Cl 1 and IV Cl 3) and one less malignant (IV Cl 2) clone. Clone IV Cl 1 was inoculated intracardially in nude mice and gave rise to adrenal and brain metastases. Lines V and VI were derived from such metastases. Multiple structural and/or numerical chromosome abnormalities were detected in all lines. Line I had 57-61 chromosomes, with structural changes affecting 1p, 2p, 3q, 7p, 7q, 11p, 14q, 17q, and 22q, as well as one unidentified marker. Line II had 40-48 chromosomes, with structural changes of 1p, 1q, 4q, 5p, 6p, 8p, 11p, 11q, 14p, 20p, and two unidentified markers. Line III had 45 chromosomes, 6q+, del(11p), and a centric fusion between chromosomes 14 and 15. Line IV had 45-46 chromosomes. The clonal changes included rearrangements of 1p, 9p, 11p, and the centric fusion of chromosomes 14 and 15. Line V was pseudodiploid and contained aberrations of 1p, 9p, 11p, 14q, 20q, an isochromosome for 21q, and an unidentified marker. Finally, the pseudodiploid line VI had changes of 9p, 11p, centric fusion of chromosomes 14 and 15, and an unidentified marker. Although no single identical aberration was shared by all six lines, structural abnormalities of 11p were invariably present and, hence, might constitute a common cytogenetic feature in melanoma development. The most consistent difference between the amelanotic and melanotic lines derived from case 3 was the presence of a 6q+ marker in the former and a 9p+ marker in the latter.

Pseudohypoparathyroidism type I and Albright's hereditary osteodystrophy with a proximal 15q chromosomal deletion in mother and daughter.

A 33-year-old woman and her 71-year-old mother were both found to have pseudohypoparathyroidism type I with Albright's hereditary osteodystrophy associated with a cytogenetic deletion of the proximal part of one chromosome 15, resembling that found in Prader-Willi syndrome. As there are overlapping clinical features between these two syndromes a causal relationship cannot be excluded. However, molecular analyses with 10 probes from this region did not detect any uniparental disomy or deletion, features frequently found in Prader-Willi syndrome.

In situ hybridization localizes the human type II alpha 1 collagen gene (COL2A1) to 12q13.

We have, using in situ hybridization technique, localized the human type II alpha 1 collagen gene (COL2A1) to chromosome band 12q13. The gene had previously been assigned to either 12q13.1-13.2 or 12q14.3. Since the chromosome segment 12q13-15 has been shown to be rearranged in several benign and malignant human neoplasms, the exact band localization of COL2A1 within this region makes it a useful marker for the molecular analysis of these tumors.

Chromosome localization of the human oncogene INT1 to 12q13 by in situ hybridization.

The human oncogene INT1 has been mapped to chromosome band 12q13 by in situ hybridization. The precise localization of this gene is of particular interest, since the region 12q13----q14 has been reported to be involved in chromosomal rearrangements in lipomas, myxoid liposarcomas, pleomorphic adenomas, and myomas. The involvement of this region in both benign and malignant tumors suggests a common pathogenetic pathway in which changes affecting INT1 may be an important step.

Chromosomal rearrangements in chondromatous tumors.

Short-term cultures from 16 chondromatous tumors, 15 primary and one recurrent, were analyzed cytogenetically. Clonal chromosome aberrations were found in one of six benign tumors and in seven of ten malignant tumors. A chondroma had a complex translocation involving chromosomes X, 8, 12, and 13, as well as a deletion of the derivative chromosome 8. In the malignant tumors, monosomy 6 and 22 were observed in three tumors and monosomy 10, 11, 13, and 18 were observed in two tumors. In two of the three metastasizing tumors, del(5) (q13) and loss of chromosomes 6, 10, 11, 13, and 22 were common features. Structural aberrations of chromosome 1 were found in five tumors, of chromosomes 6, 12, and 15 in three tumors, and of chromosomes 4, 5, 9, and 20 in two tumors. We conclude that although considerable cytogenetic heterogeneity exists among chondromatous tumors, the karyotypic anomalies are still nonrandom.

Deletions of CDKN1B and ETV6 in acute myeloid leukemia and myelodysplastic syndromes without cytogenetic evidence of 12p abnormalities.

Seventy-nine acute myeloid leukemias (AML) and myelodysplastic syndromes without cytogenetic evidence of 12p aberrations were investigated by fluorescence in situ hybridization with probes for ETV6 and CDKN1B (previously called TEL and KIP1, respectively) to ascertain whether abnormalities of these genes are frequently undetected by standard chromosome banding analyses and, if so, whether they are associated with specific karyotypic patterns and morphologic features. One of sixty cytogenetically aberrant myeloid malignancies, an AML with a complex karyotype including del(5q) and del(20q), showed a hemizygous interstitial deletion of the ETV6 and CDKN1B loci. No concomitant rearrangement of the other ETV6 allele was detected. Two of nineteen cytogenetically normal AML displayed a hemizygous interstitial deletion involving CDKN1B, but not ETV6. Thus, cryptic deletions of these genes seem to be rare in cytogenetically abnormal myeloid malignancies without 12p aberrations (2%), whereas they may be more frequent in karyotypically normal AML (10%). Furthermore, the present findings show that the deletions may be narrow, not including the ETV6 gene, and indirectly suggest that CDKN1B, or a closely located genomic segment, is the target of 12p deletions.

Localization in man of fifteen DNA sequences within the chromosome segment 13q12-q22.

Fifteen human chromosome 13 specific DNA fragments, isolated from a lambda phage genomic library, were localized within the segment 13q12-q22. One was mapped to 13q12.1-q12.2, three to 13q12.3-q13.1, one to 13q14,1-q14.2, five to 13q14.1-q21.1, one to 13q21.1-q21.2, two to 13q21.2, and one to 13q22.1, and one to 13q22. The localization was performed by hybridization to Southern blots of a panel of human cell lines with overlapping deletions in 13q, and for three probes also by in situ hybridization to metaphase chromosomes.

Three major cytogenetic subgroups can be identified among chromosomally abnormal solitary lipomas.

We have investigated cytogenetically a total of 35 solitary lipomas, 10 of which have been reported previously. Of the 25 tumours presented herein for the first time, clonal chromosome aberrations were detected in 17. The remaining eight had normal karyotypes, although two of them had nonclonal aberrations in about one quarter of the cells. Based on the cytogenetic findings in all 35 lipomas, four major subgroups can be distinguished. These are characterized by: (I) hyperdiploid karyotypes including one or more supernumerary ring chromosomes (5 cases); (II) diploid karyotypes with mostly balanced rearrangements involving 12q13-14 (13 cases), including the rearrangement t(3;12) (q27-28;q13-14) in 4 cases; (III) hypodiploid or diploid karyotypes with other aberrations than ring chromosomes or rearrangements of 12q13-14 (8 cases); and (IV) normal karyotypes (9 cases).

Rearrangement of the transcription factor gene CHOP in myxoid liposarcomas with t(12;16)(q13;p11).

Most myxoid liposarcomas (MLS) are characterized cytogenetically by a t(12;16)(q13;p11). It is reasonable to assume that this translocation corresponds to the consistent rearrangement of one or two genes in 12q13 and/or 16p11, and that the loci thus affected are important in the normal control of fat cell differentiation and proliferation. We have used Southern blot technique to test whether a gene of the CCAAT/enhancer binding protein (C/EBP) family, CHOP, which maps to 12q13 and is assumed to be involved in adipocyte differentiation, could be the 12q gene in question. Using a cDNA probe that spans the CHOP coding region, we detected one rearranged and one wild type allele in nine of nine MLS with t(12;16). Using PCR generated, site-specific probes corresponding to the non-coding exons 1 and 2 and intron 2 of CHOP, rearrangements in five of seven tumors mapped to the 2.4 and 1.6 kbp PstI fragments that contain the first two exons and introns of the gene and the upstream promoter region. In contrast to the findings in MLS, no tumor without a t(12;16) exhibited aberrant CHOP restriction digest patterns. These tumors included one highly differentiated liposarcoma with abnormal karyotype but no involvement of 12q13, seven lipomas with various cytogenetic aberrations of 12q13-15, two uterine leiomyomas with t(12;14) (q14-15;q23-24), and one hemangiopericytoma and one chondroma, both of which also had 12q13 changes.(ABSTRACT TRUNCATED AT 250 WORDS)

Karyotypic features of malignant tumors of the nasal cavity and paranasal sinuses.

Cytogenetic analysis of short-term cultures from 6 tumors of the nasal cavity and paranasal sinuses--one esthesioneuroblastoma, 2 adenocarcinomas and 3 squamous-cell carcinomas (SCC)--revealed clonal chromosome aberrations in all cases. The esthesioneuroblastoma had a complex hyperdiploid karyotype. None of the aberrations was similar to those previously described in short-term cultures or established cell lines from esthesioneuroblastomas. The 2 adenocarcinomas had complex karyotypic changes, which in both cases included rearrangements of bands 9p22 and 14q11. One SCC had 5 unrelated pseudodiploid clones, 1 displayed a highly complex karyotype, including rearrangement of band 11q13, and 1 had simple karyotypic changes with loss of 6q material and gain of 3q. These findings are similar to those described in head-and-neck SCC at other sites.