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Suppression of vascular endothelial growth factor via siRNA interference modulates the biological behavior of human nasopharyngeal carcinoma cells.
Japanese Journal of Radiology 2011 November
PURPOSE: The aim was to study the effect of vascular endothelial growth factor (VEGF) down-regulation by small interfering (si)RNA-mediated interference (RNAi) on the biological features of nasopharyngeal carcinoma cell line CNE-2.
MATERIALS AND METHODS: The combined plasmids pU-siVEGF and pU-siCONT were transfected into CNE-2 cells with lipofectamine. The transfected cells were placed in fresh medium containing G418. Expression of VEGF mRNA and protein were measured by reverse transcriptase-polymerase chain reaction and Western blot, respectively. The transwell chamber model was employed to test the ability of cell invasion in vitro. The distribution of cell cycle phases was determined by flow cytometry. Cell survival was assessed by clonogenic assays.
RESULTS: Both VEGF mRNA and protein expression were significantly decreased in the pU-siVEGF group compared with controls (P < 0.05). The cell cycle was arrested in the G(1) phase (P < 0.05). A higher apoptotic ratio and lower invasion ability were seen in the pU-siVEGF group. The D(0) (mean lethal dose) and SF(2) values were significantly lower than those in the control group (P < 0.05).
CONCLUSION: Delivery of siRNA targeting VEGF seems efficient in down-regulating VEGF expression and diminishing the growth, proliferation, and invasiveness of CNE-2 cells. It also enhanced the sensitivity of CNE-2 cells to radiation.
MATERIALS AND METHODS: The combined plasmids pU-siVEGF and pU-siCONT were transfected into CNE-2 cells with lipofectamine. The transfected cells were placed in fresh medium containing G418. Expression of VEGF mRNA and protein were measured by reverse transcriptase-polymerase chain reaction and Western blot, respectively. The transwell chamber model was employed to test the ability of cell invasion in vitro. The distribution of cell cycle phases was determined by flow cytometry. Cell survival was assessed by clonogenic assays.
RESULTS: Both VEGF mRNA and protein expression were significantly decreased in the pU-siVEGF group compared with controls (P < 0.05). The cell cycle was arrested in the G(1) phase (P < 0.05). A higher apoptotic ratio and lower invasion ability were seen in the pU-siVEGF group. The D(0) (mean lethal dose) and SF(2) values were significantly lower than those in the control group (P < 0.05).
CONCLUSION: Delivery of siRNA targeting VEGF seems efficient in down-regulating VEGF expression and diminishing the growth, proliferation, and invasiveness of CNE-2 cells. It also enhanced the sensitivity of CNE-2 cells to radiation.
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