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1.
Oncogene ; 9(6): 1649-53, 1994 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-8183559

RESUMO

Twenty microsatellite loci on chromosome 9 were analysed for allelic losses in DNAs from 30 uncultured melanomas from 25 patients, relative to DNA from autologous peripheral blood lymphocytes. All patients were constitutionally heterozygous at several loci, and loss of heterozygosity (LOH) affecting 9p was observed in melanoma DNAs from 18 individuals (72%). Observations of losses of identical alleles in different metastatic lesions from the same patients, and of LOH in a vertical growth phase primary melanoma, were consistent with previous reports of chromosome 9 deletion early in melanoma development. LOH data suggested the loss of entire copies of chromosome 9 in 11 cases, and the terminal deletion of all or a portion of 9p in six cases. A somatic interstitial deletion of 9p between D9S162 and D9S169 was seen in a familial melanoma. This 21 cM deleted region corresponded with the previously reported positions of homozygous deletions in melanoma cell lines, and of the familial melanoma susceptibility locus (MLM). As 16 of the 18 cases of 9p LOH in the present study were observed in individuals with no family history of melanoma, it is likely that the MLM locus plays a role in the development of most sporadic melanomas.


Assuntos
Deleção Cromossômica , Cromossomos Humanos Par 9 , Melanoma/genética , Mapeamento Cromossômico , DNA de Neoplasias/análise , Humanos , Reação em Cadeia da Polimerase
3.
Hum Mol Genet ; 2(9): 1407-11, 1993 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-7902161

RESUMO

A panel of 18 superficial or invasive transitional cell carcinomas (TCCs) was analyzed for chromosome 9 deletions by performing a high-density loss of heterozygosity (LOH) analysis. Twenty-five microsatellite loci were assayed by the polymerase chain reaction (PCR) and 7 restriction fragment length polymorphism (RFLP) loci were analyzed by Southern blotting. Concordant results were obtained with these methods, including direct comparisons at 2 loci. Chromosome 9 LOH was observed in 13 (72%) of 18 informative cases, including 10 superficial lesions. In contrast, LOH on chromosomes 10, 15, 20 and 21 was < or = 8%. Evidence for missing copies of chromosome 9 was observed in 7 of 13 LOH cases. Comparison of cases with subchromosomal LOH implicated the region between the D9S55 locus on 9p12 and the argininosuccinate synthetase (ASS) locus on 9q34.1 as the location of a tumor suppressor gene relevant to TCC.


Assuntos
Carcinoma de Células de Transição/genética , Deleção Cromossômica , Cromossomos Humanos Par 9 , Neoplasias da Bexiga Urinária/genética , Argininossuccinato Sintase/genética , Sequência de Bases , Primers do DNA/genética , DNA de Neoplasias/genética , DNA Satélite/genética , Marcadores Genéticos , Humanos , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de Restrição
4.
Mol Cell Biol ; 13(3): 1507-15, 1993 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-8382772

RESUMO

The gene encoding the carcinoma-associated antigen defined by the monoclonal antibody GA733 is a member of a family of at least two type I membrane proteins. This study describes the mechanism of evolution of the GA733-1 and GA733-2 genes. A full-length cDNA clone for GA733-1 was obtained by screening a human placental library with a genomic DNA probe. Comparative analysis of the cDNA sequence with the previously determined genomic sequence confirmed that GA733-1 is an intronless gene. The GA733-2 gene encoding the monoclonal antibody-defined antigen was molecularly cloned with a cDNA probe and partially sequenced. Comparison of GA733-2 gene sequences with the previously established cDNA sequence revealed that this gene consists of nine exons. The putative promoter regions of the GA733-1 and GA733-2 genes are unrelated. These findings suggest that the GA733-1 gene was formed by the retroposition of the GA733-2 gene via an mRNA intermediate. Prior to retroposition, the GA733-2 gene had been affected by exon shuffling. Analysis of GA733-2 exons revealed that many delineate structural motifs. The GA733-1 retroposon was localized either to chromosome region 1p32-1p31 or to 1p13-1q12, and the GA733-2 founder gene was localized to chromosome 4q.


Assuntos
Antígenos Glicosídicos Associados a Tumores/genética , Evolução Biológica , Elementos de DNA Transponíveis/genética , Sequência de Aminoácidos , Sequência de Bases , Mapeamento Cromossômico , Cromossomos Humanos Par 1 , Cromossomos Humanos Par 4 , Clonagem Molecular , Genoma Humano , Humanos , Dados de Sequência Molecular , Família Multigênica , RNA Mensageiro/biossíntese , Análise de Sequência de DNA , Homologia de Sequência de Aminoácidos
5.
Cancer Res ; 52(4): 815-21, 1992 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-1737342

RESUMO

GA733-2 is a monoclonal antibody-defined, 40-kDa glycoprotein antigen that is associated with carcinomas of various origins. Hydrophobicity analysis of the protein sequence predicted by complementary DNA (cDNA) has suggested that the GA733-2 antigen is a type I membrane protein. In this study, the polymerase chain reaction was used in a strategy to omit cDNA sequences for the transmembrane and cytoplasmic domains, thereby converting the extracellular domain into a secretory protein. Full-length and truncated cDNAs were cloned into the baculovirus transfer vector pVL1392 and introduced into Autographa californica nuclear polyhedrosis virus by homologous recombination. The full-length cDNA baculovirus recombinant directed the expression of a 40-kDa glycoprotein that was confined to infected Spodoptera frugiperda cells, whereas cells infected with the truncated cDNA baculovirus recombinant abundantly secreted a 31-kDa glycoprotein into the culture medium. Recombinant secretory antigen displayed an in vitro immunoreactivity to monoclonal antibody and an in vivo immunogenicity in mice that were similar to native antigen. The facile purification of mg quantities of carcinoma-associated antigen will enable an evaluation of its immunogenicity in cancer patients.


Assuntos
Antígenos de Neoplasias/genética , Baculoviridae/genética , Moléculas de Adesão Celular , Transfecção , Animais , Anticorpos Monoclonais , Antígenos de Neoplasias/análise , Antígenos de Neoplasias/imunologia , Sequência de Bases , Linhagem Celular , Clonagem Molecular , Molécula de Adesão da Célula Epitelial , Vetores Genéticos , Humanos , Proteínas de Membrana/genética , Dados de Sequência Molecular , Mariposas , Oligodesoxirribonucleotídeos , Reação em Cadeia da Polimerase/métodos , Proteínas Recombinantes/análise , Recombinação Genética , Mapeamento por Restrição
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