RESUMO
NF-?B, a group of important transcription factors, are introduced and discussed here on several aspects: their component and molecular structure; their activity control by I?B and IKK, the mechanism of their activation; their important roles in transcriptional regulation for large numbers of genes; and their importance in immunity, inflammation, cell survival, proliferation and apoptosis. The analysis of the relation between NF-?B and disease occurrence, the analysis of the relation between NF-?B and disease therapy, and the application prospect of the new strategy regarding the novel drug design and correlative diseases therapy on the basis of NF-?B as the target, are also included.
RESUMO
<p><b>OBJECTIVE</b>To establish a protocol for the targeting gene therapy against cancer with rich epidermal growth factor receptor (EGFR).</p><p><b>METHODS</b>A recombinant pcDNA3.1-PE III mut was constructed and combined with a non-viral vector, a fusion protein histone H1, epidermal growth factor C-loop previously expressed by us, to be a protein-DNA complex in vitro. Using the complex to treat BT-325 and Hela cancer cells with EGFR and JK cells without EGFR. The killing rates of the cells was calculated after 48 h of incubation at 37 degrees C.</p><p><b>RESULTS</b>To BT-325 and Hela cells, the killing rates were 46.03% and 48.12% respectively. To JK cells, the complex had no killing function.</p><p><b>CONCLUSION</b>The protocol for targeting gene therapy against cancer with EGFR has been established successfully.</p>
Assuntos
Humanos , ADP Ribose Transferases , Genética , Farmacologia , Toxinas Bacterianas , Genética , Farmacologia , Sequência de Bases , Linhagem Celular Tumoral , Células , DNA , Genética , Exotoxinas , Genética , Farmacologia , Marcação de Genes , Terapia Genética , Vetores Genéticos , Histonas , Genética , Dados de Sequência Molecular , Receptores ErbB , Genética , Metabolismo , Proteínas Recombinantes de Fusão , Genética , Metabolismo , Farmacologia , Transfecção , Fatores de Virulência , Genética , FarmacologiaRESUMO
<p><b>OBJECTIVE</b>To create a gene transfer vehicle for targeting gene therapy of cancer with epidermal growth factor receptor overexpressing.</p><p><b>METHODS</b>Encoding sequences of the first domain of histone gene (H1) and EGF C-loop (EGFc) were obtained by PCR amplification. These two DNA fragments were ligated by EcoR I site, and cloned and sequenced. E. coli expression vector of the fusion gene was then constructed. The fusion protein H1EGFc was purified by specific band isolation from SDS-PAGE.</p><p><b>RESULTS</b>The molecular weight of purified protein was consistent with the designed request. Its purity reached 94.02%.</p><p><b>CONCLUSION</b>A fusion protein H1EGFc was expressed and purified.</p>
Assuntos
Humanos , Sequência de Aminoácidos , Sequência de Bases , Escherichia coli , Genética , Técnicas de Transferência de Genes , Terapia Genética , Vetores Genéticos , Histonas , Genética , Dados de Sequência Molecular , Engenharia de Proteínas , Receptores ErbB , Genética , Proteínas Recombinantes de Fusão , GenéticaRESUMO
In order to explore the feasibility of gene therapy strategy based on the human atrial natriuretic peptide (hANP) gene delivery for the treatment of nephropathy and compare the diuretic activities of the hANP gene injected intramuscularly(i.m.) and intravenously(i.v.), the naked retroviral vector DNA harboring the hANP cDNA under the control of retroviral 5' long terminal repeat at a dose of 5 mg/kg body weight was injected i.m. or i.v. into the nephrotic model rats induced with adriamycin(ADR) injected i.v. at a dose of 7.5 mg/kg body weight. A single injection of the hANP gene resulted in a marked elevation in plasma level of hANP 5 days after gene delivery and a significant increase in the ratio of urine volume to body weight and the diuretic effect continued for more than 15 days. In addition, there was a significant rise in the body weight of treatment groups as compared with that of negative control group and no difference in the concentrations of electrolytes in urine between groups. There was no significant differences in total effects resulted from the two routes of gene delivery and the way of gene delivery through the skeletal muscle is simpler and easier. These results suggest that somatic gene delivery of the hANP gene could enhance the renal functions in nephrotic rats significantly and would be a potential strategy for the treatment of renal disorders.