Effects of RNA interference on epidermal growth factor receptor expression in breast cancer cells: a study in tumor-bearing nude mice / 南方医科大学学报
Journal of Southern Medical University
; (12): 60-64, 2008.
Article
en Zh
| WPRIM
| ID: wpr-281480
Biblioteca responsable:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of cationic liposome-mediated RNA interference (RNAi) in silencing epidermal growth factor (EGF) receptor (EGFR) gene in breast cancer cells in vivo.</p><p><b>METHODS</b>A small interfering RNA (siRNA) targeting EGFR gene was constructed and transfected into human breast cancer cell in vitro via cationic liposome. The transfected cells were inoculated into nude mice, and the tumor growth inhibition rate was calculated. The tumors were then removed for immunohistochemistry and Western blotting to examine the expression of EGFR protein. Quantitative RT-PCR was used to detect the mRNA expression of the EGF receptor gene, and enzyme-linked immunosorbent assay (ELISA) performed to assess the EGF level in both the serum and tumor extraction.</p><p><b>RESULTS</b>In athymic nude mice, MDA-MB-231 cells had obviously lower tumor formation rate than ZR-75 cells (30.00% and 88.89%). Transfection of the cells with EGFR siRNA significantly inhibited tumor formation capacity of the cells in vivo as compared with the cells transfected with empty vector or irrelevant siRNA. The results of ELISA demonstrated that in mice bearing the tumors grown from EGFR siRNA-transfected cells, the EGF levels in the serum and tumor extraction were lowered by 16.77% and 12.59%, respectively. Real-time RT-PCR showed that EGFR siRNA transfection caused a specific downregulation of EGFR mRNA expression by 21.05% in the tumor.</p><p><b>CONCLUSION</b>Chemically synthesized 21-nucleotide siRNA duplexes can be effectively delivered via lipofectamine 2000 into breast cancer cells in vivo to induce a longer-lasting gene silencing effect than in vitro transfection. RNAi of EGFR gene may indicate a promising approach for management of lung cancers, especially those nodular ones with easy access.</p>
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1
Base de datos:
WPRIM
Asunto principal:
Patología
/
Terapéutica
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Neoplasias de la Mama
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Ensayo de Inmunoadsorción Enzimática
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Transfección
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Distribución Aleatoria
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Regulación Neoplásica de la Expresión Génica
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Reacción en Cadena de la Polimerasa de Transcriptasa Inversa
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Ensayos Antitumor por Modelo de Xenoinjerto
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ARN Interferente Pequeño
Tipo de estudio:
Prognostic_studies
Límite:
Animals
Idioma:
Zh
Revista:
Journal of Southern Medical University
Año:
2008
Tipo del documento:
Article