Title: Inhibition of Malignant Melanoma Cell Growth Through Blocking Expression of VEGF by RNA Interference
Abstract: Objectives To construct a small interfering RNA (siRNA) targeting vascular endothelial growth factor (VEGF) in eukaryotic expression vector, and to assess effects of this vector on proliferation and apoptosis of malignant melanoma cell line A375. Methods pU-VEGF-siRNA plasmid was transfected into A375 cells by electroporation. VEGF mRNA and protein were detected by reverse transcription polymerse chain reaction (RT-PCR) and ELISA, respectively, in A375 cells after gene transfer. Proliferation of A375 cells was assessed by cell counting. Human umbilical vein endothelial cells (ECV-304) were cultured in medium containing supernatants of treated and untreated A375 cells, and their viability was tested by cell counting. Apoptosis of A375 cells was observed by flow cytometry (FCM). Results VEGF mRNA and protein were significantly decreased in the experimental group, compared to those in the controls (P 0.05). Proliferation of treated A375 and ECV-304 cells was inhibited. Subdiploidy peaks were also found in the treated A375 cells. Conclusions Delivery of siRNA targeting VEGF seems efficient in down-regulating VEGF expression, in inhibiting A375 and ECV-304 cell proliferation and in inducing A375 cell apoptosis, suggesting that siRNA-based strategy targeting VEGF strategy may lay a foundation for clinical management of malignant melanoma.
Publication Year: 2005
Publication Date: 2005-01-01
Language: en
Type: article
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