[Suppression of vascular endothelial growth factor expression by vector-based small interfering RNA in human tongue squamous carcinoma cell line Tca8113].
Dahai Yu, Ying Cao, Zhiwen Yao, Jing Li, Haibo Chen, Jie Hao
Abstract
Dahai Yu, Ying Cao, Zhiwen Yao, Jing Li, Haibo Chen, Jie Hao
Abstract
OBJECTIVE: To assess suppression effects of vector-based small interfering RNA (siRNA) on vascular endothelial growth factor (VEGF) expression of human tongue squamous carcinoma cell line (Tca8113) in vitro. METHODS: Two siRNA targeting VEGF constructed in eukaryotic expression vector (Pu-VEGF-siRNA1, Pu-VEGF-siRNA2), eukaryotic expression vector as the experiment control, all of which were transfected into Tca8113 cells with Lipofectamine 2000. Non-transfection cell was used as negative control. VEGF mRNA and protein were detected by reverse transcription polymerase chain reaction (RT-PCR), immunohistochemistry and enzyme linked immunosorbent assay (ELISA), respectively. RESULTS: Compared to the experimental and negative controls, the expression of VEGF mRNA and protein were significantly decreased in the Pu-VEGF-siRNA1 group and Pu-VEGF-siRNA2 group. But there were no significant differences between two controls (P > 0.05). CONCLUSION: Vector-based siRNAs targeting VEGF are efficient in down-regulating VEGF expression in Tca8113 cells.
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OBJECTIVE: To assess suppression effects of vector-based small interfering RNA (siRNA) on vascular endothelial growth factor (VEGF) expression of human tongue squamous carcinoma cell line (Tca8113) in vitro. METHODS: Two siRNA targeting VEGF constructed in eukaryotic expression vector (Pu-VEGF-siRNA1, Pu-VEGF-siRNA2), eukaryotic expression vector as the experiment control, all of which were transfected into Tca8113 cells with Lipofectamine 2000. Non-transfection cell was used as negative control. VEGF mRNA and protein were detected by reverse transcription polymerase chain reaction (RT-PCR), immunohistochemistry and enzyme linked immunosorbent assay (ELISA), respectively. RESULTS: Compared to the experimental and negative controls, the expression of VEGF mRNA and protein were significantly decreased in the Pu-VEGF-siRNA1 group and Pu-VEGF-siRNA2 group. But there were no significant differences between two controls (P > 0.05). CONCLUSION: Vector-based siRNAs targeting VEGF are efficient in down-regulating VEGF expression in Tca8113 cells.
Key concepts: Small interfering RNA, Lipofectamine, Vascular endothelial growth factor, Transfection, Molecular biology, Messenger RNA, Expression vector, Cell culture