Inhibition of Malignant Melanoma Cell Growth Through Blocking Expression of VEGF by RNA Interference / 中华皮肤科杂志

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