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>