Targeted inhibition of vascular endothelial growth factor (VEGF) expression in human osteosarcoma cell line by antisense VEGF165 cDNA promoted by hypoxia reaction element / 中华病理学杂志
Chinese Journal of Pathology
; (12): 588-591, 2005.
Artigo
em Chinês
| WPRIM (Pacífico Ocidental)
| ID: wpr-297285
Biblioteca responsável:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>Utilizing the hypoxia inducible factor 1/hypoxia reaction element (HIF-1/ HRE) gene regulation system to construct antisense vascular endothelial growth factor (VEGF165) cDNA eukaryotic expression vector promoted by HRE, and investigate its targeted inhibiting VEGF expression of osteosarcoma cells in hypoxia environment.</p><p><b>METHODS</b>Eukaryotic expression plasmid with HRE promoter was constructed containing luciferase reporter gene and antisense VEGF165 cDNA by using PCR and recombinant DNA techniques. The recombinant vectors were transfected into osteosarcoma cells with lipofectin method. Hypoxia-inducible reporter gene expression was determined by liquid scintillation analyzer and the expression of VEGF protein was detected by ELISA method.</p><p><b>RESULTS</b>The eukaryotic expression plasmid containing antisense VEGF165 and luciferase promoted by HRE was constructed successfully. After being transferred into MG63 cells, luciferase expression was increased 3.5 x 10(2) times and VEGF protein expression decreased 45% under hypoxia condition.</p><p><b>CONCLUSION</b>Antisense VEGF165 cDNA expression, efficiently realized by HRE promoter under hypoxia condition, provides an experimental basis for targeted antiangiogenesis of tumors.</p>
Texto completo:
Disponível
Base de dados:
WPRIM (Pacífico Ocidental)
Assunto principal:
Patologia
/
Plasmídeos
/
Neoplasias Ósseas
/
Proteínas Recombinantes
/
Transfecção
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Osteossarcoma
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Hipóxia Celular
/
Regiões Promotoras Genéticas
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Oligodesoxirribonucleotídeos Antissenso
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Linhagem Celular Tumoral
Limite:
Humanos
Idioma:
Chinês
Revista:
Chinese Journal of Pathology
Ano de publicação:
2005
Tipo de documento:
Artigo