Assuntos
Anticorpos/imunologia , Rim Policístico Autossômico Dominante/genética , Proteínas/genética , Animais , Western Blotting , Linhagem Celular , Eletroforese em Gel de Poliacrilamida , Escherichia coli/genética , Expressão Gênica , Humanos , Imuno-Histoquímica , Neoplasias Renais/química , Neoplasias Renais/patologia , Fragmentos de Peptídeos/química , Fragmentos de Peptídeos/genética , Fragmentos de Peptídeos/imunologia , Reação em Cadeia da Polimerase , Proteínas/química , Proteínas/imunologia , RNA Mensageiro/análise , RNA Mensageiro/genética , Coelhos , Proteínas Recombinantes de Fusão/biossíntese , Proteínas Recombinantes de Fusão/imunologia , Proteínas Recombinantes de Fusão/isolamento & purificação , Canais de Cátion TRPP , Células Tumorais CultivadasRESUMO
A group of 20 CLL patients selected for advanced clinical stage p53 mutations were analysed by single-strand conformational polymorphism (SSCP) following PCR amplification of exons 5-9. In two patients abnormal SSCP of either exon 5 or exon 8 was found and PCR products were analysed by direct sequencing. A hemizygous or homozygous 12bp deletion at codon 135 and 3bp heterozygous deletion at codon 264 were detected; also, in the latter sample a heterozygous mutation at codon 282 (Arg to Gln) was found. To our knowledge, this is the first report of p53 deletions in B-CLL. The two patients were elderly, and both had a rapidly progressive disease in the absence of unfavourable cytogenic abnormalities. These findings support a role for p53 alterations in the clinical course of some B-CLL patients.
Assuntos
Genes p53/genética , Leucemia Linfocítica Crônica de Células B/genética , Deleção de Sequência , Idoso , Sequência de Bases , Feminino , Humanos , Dados de Sequência Molecular , Mutação Puntual , Reação em Cadeia da Polimerase , Polimorfismo Conformacional de Fita SimplesRESUMO
Loss of chromosome 6q was investigated in endometrial and ovarian carcinomas by a PCR based-microsatellite polymorphism analysis. Results obtained show that this technique is able to detect frequent loss of heterozygosity in the ovarian cancers (10/27) and only in the serous type (8/17). Then, this kind of analysis can contribute to the understanding of tumor development and progression of ovarian cancers with different histopathological features.
Assuntos
Deleção Cromossômica , Cromossomos Humanos Par 6 , DNA de Neoplasias , DNA Satélite , Neoplasias Ovarianas/genética , Polimorfismo Genético , Sequência de Bases , Primers do DNA/química , Feminino , Heterozigoto , Humanos , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Receptores de Estrogênio/genéticaRESUMO
The effects of androgen and estrogen on cell growth and gene expression were investigated in KJ29 kidney epithelial cells. Incorporation of 3H leucine and 3H thymidine was increased by androgen at 10nM, but not by estrogen. Estrogen however, inhibited the effects induced by androgen. In addition, cell number and the proliferation marker Ki67 were increased by androgen, but not by estrogen. Levels of androgen receptor RNA, as detected by RT-PCR and Northern blot analysis, were not affected by either androgen or estrogen. Levels of estrogen receptor RNA could be detected only by RT-PCR, and disappeared after estrogen treatment. These studies show that sex steroid receptors are differently expressed in KJ29 cells, and suggest that androgen, via its canonic receptor, acts as a mitogenic factor in human kidney cells, whereas estrogen has an antiandrogenic action.
Assuntos
Estradiol/farmacologia , Expressão Gênica , Rim/metabolismo , Receptores Androgênicos/biossíntese , Testosterona/farmacologia , Anticorpos Monoclonais , Sequência de Bases , Northern Blotting , Divisão Celular/efeitos dos fármacos , Linhagem Celular , Primers do DNA , Relação Dose-Resposta a Droga , Epitélio/efeitos dos fármacos , Epitélio/metabolismo , Expressão Gênica/efeitos dos fármacos , Humanos , Neoplasias Renais , Cinética , Leucina/metabolismo , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Receptores de Estrogênio/análise , Receptores de Estrogênio/biossíntese , Timidina/metabolismo , Fatores de TempoRESUMO
Two transcripts of the human estrogen receptor (ER) gene have been described, ER mRNA 1 and mRNA 2, different in their 5' untranslated region. By performing reverse transcriptase-polymerase chain reaction with oligonucleotides specific for the 5' genomic region of the human ER gene we have identified a new ER RNA transcript. The sequence analysis of cDNA from MCF7 breast cancer cells and endometrial human tissues demonstrates that this transcript originates further upstream of the initiation transcription sites so far proposed. Primer extension analysis on RNA from MCF7 cells reveals in the upstream region a possible transcription start site at -3090. In agreement with this result, Northern blot analysis shows, in addition to the canonical 6.3 kb ER mRNA, an ER RNA transcript of approx. 7.4 kb in size. The presence of the additional ER mRNA suggests the existence of a new upstream 5' promoter directing transcription of the human ER gene.
Assuntos
RNA Mensageiro/genética , Receptores de Estrogênio/genética , Transcrição Gênica , Sequência de Bases , Neoplasias da Mama , Primers do DNA , Sondas de DNA , Endométrio , Éxons , Feminino , Humanos , Íntrons , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Análise de Sequência de DNA , Células Tumorais CultivadasRESUMO
In order to demonstrate that Androgen binding activity in thyroid is caused by the canonic Androgen Receptor (AR), member of steroid receptor family, we studied the presence of AR mRNA in human thyroid tissues and primary cultured cells. Here we report a polymerase chain reaction protocol (RT-PCR) that we have designated to investigate the presence of AR mRNA in human cells. AR cDNA was synthesized and amplified with primers specific for C-terminal sequence of the protein. We demonstrated that AR gene expression i) is present in thyroid samples studied and ii) in a primary culture of follicular adenoma where it seems to be modulated by steroid hormones.