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Inhibition effects of RNAi on VEGF-mRNA expression in HCT116 cells

Wei Lü

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Abstract

Objective To investigate the effects of RNAi on VEGF-mRNA expression in HCT116 cells. Methods The Pavu6+27-VEGF siRNA expression vector was constructed by gene recombination, then transfected into the cultured HCT116 cells by DOTAP method. The positive cell clones were screened with G418. The stable transfection and expression of VEGF-mRNA in HCT116 were determined by semi-quantitative RT-PCR. The Pavu6+27 transfected plasmid was used as control. Results Pavu6+27-VEGF siRNA expression vector was successfully constructed. DOTAP-mediated gene transfection of Pavu6+27-VEGF siRNA expression vector into HCT116 cells down-regulated the mRNA expression level of VEGF gene, as compared with the control group. Conclusion The Pavu6+27-VEGF siRNA expression vector can inhibit the expression of VEGF-mRNA in HCT116 cells, which may be beneficial in searching new gene therapy of the tumors.

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Objective To investigate the effects of RNAi on VEGF-mRNA expression in HCT116 cells. Methods The Pavu6+27-VEGF siRNA expression vector was constructed by gene recombination, then transfected into the cultured HCT116 cells by DOTAP method. The positive cell clones were screened with G418. The stable transfection and expression of VEGF-mRNA in HCT116 were determined by semi-quantitative RT-PCR. The Pavu6+27 transfected plasmid was used as control. Results Pavu6+27-VEGF siRNA expression vector was successfully constructed. DOTAP-mediated gene transfection of Pavu6+27-VEGF siRNA expression vector into HCT116 cells down-regulated the mRNA expression level of VEGF gene, as compared with the control group. Conclusion The Pavu6+27-VEGF siRNA expression vector can inhibit the expression of VEGF-mRNA in HCT116 cells, which may be beneficial in searching new gene therapy of the tumors.

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Available abstract

Objective To investigate the effects of RNAi on VEGF-mRNA expression in HCT116 cells. Methods The Pavu6+27-VEGF siRNA expression vector was constructed by gene recombination, then transfected into the cultured HCT116 cells by DOTAP method. The positive cell clones were screened with G418. The stable transfection and expression of VEGF-mRNA in HCT116 were determined by semi-quantitative RT-PCR. The Pavu6+27 transfected plasmid was used as control. Results Pavu6+27-VEGF siRNA expression vector was successfully constructed. DOTAP-mediated gene transfection of Pavu6+27-VEGF siRNA expression vector into HCT116 cells down-regulated the mRNA expression level of VEGF gene, as compared with the control group. Conclusion The Pavu6+27-VEGF siRNA expression vector can inhibit the expression of VEGF-mRNA in HCT116 cells, which may be beneficial in searching new gene therapy of the tumors.

Key concepts: Transfection, RNA interference, Messenger RNA, Molecular biology, Expression vector, Gene expression, Small interfering RNA, Gene

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