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Construction of cell line of small hairpin RNA-mediated inhibition of HER2/neu gene expression / 中华预防医学杂志
Chinese Journal of Preventive Medicine ; (12): 714-717, 2009.
Artículo en Chino | WPRIM | ID: wpr-316109
ABSTRACT
<p><b>OBJECTIVE</b>To establish the stable inhibition of HER2/neu expression by vector-mediated small hairpin RNA in malignant transformed human bronchial epithelial cell line induced by anti-benzo(a)pyrene-trans-7, 8-dihydrodiol-9, 10-epoxide (anti-BPDE).</p><p><b>METHODS</b>The pSIREN-RetroQ-neu recombinant vector targeting HER2/neu was constructed and confirmed by restriction and sequencing analysis, then it was transfected into anti-BPDE malignant transformed 16HBE cells (16HBE-T) through lipofectamine 2000. The control groups included the 16HBE-T cells transfected with negative control vector (negative control) and 16HBE-T. The cells transfected with vectors were screened by puromycin. The HER2/neu mRNA and protein expressions in the vector-transfected 16HBE-T cells were detected by RT-PCR and Western blot method respectively.</p><p><b>RESULTS</b>The pSIREN-RetroQ-neu recombinant vector which inhibited HER2/neu mRNA and protein expressions in the 16HBE-T was constructed. The level of HER2/neu mRNA in the 16HBE-T cells transfected with pSIREN-RetroQ-neu was significantly reduced as compared to the negative control and blank control cells (0.114 +/- 0.003 vs.blank control 0.186 +/- 0.001, t = 39.154, P < 0.05; and negative control 0.182 +/- 0.015, t = 7.564, P < 0.05), while its level did not differ significantly between negative control cells and blank control of 16HBE-T (t = -0.409, P > 0.05). HER2/neu protein level in pSIREN-RetroQ-neu transfected cells was inhibited by 40% and 39% respectively.</p><p><b>CONCLUSION</b>Plasmid-based shRNA expression systems targeted against HER2/neu gene were generated successfully, which resulted in down-regulation of HER2/neu gene expression in the 16HBE-T efficiently.</p>
Asunto(s)
Texto completo: Disponible Índice: WPRIM (Pacífico Occidental) Asunto principal: 7,8-Dihidro-7,8-dihidroxibenzo(a)pireno 9,10-óxido / Línea Celular Transformada / Datos de Secuencia Molecular / Secuencia de Bases / Transfección / Expresión Génica / Genes erbB-2 / Reacción en Cadena de la Polimerasa de Transcriptasa Inversa / ARN Interferente Pequeño / Células Epiteliales Límite: Humanos Idioma: Chino Revista: Chinese Journal of Preventive Medicine Año: 2009 Tipo del documento: Artículo

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Texto completo: Disponible Índice: WPRIM (Pacífico Occidental) Asunto principal: 7,8-Dihidro-7,8-dihidroxibenzo(a)pireno 9,10-óxido / Línea Celular Transformada / Datos de Secuencia Molecular / Secuencia de Bases / Transfección / Expresión Génica / Genes erbB-2 / Reacción en Cadena de la Polimerasa de Transcriptasa Inversa / ARN Interferente Pequeño / Células Epiteliales Límite: Humanos Idioma: Chino Revista: Chinese Journal of Preventive Medicine Año: 2009 Tipo del documento: Artículo