Interference Of Human Malignant Melanoma Cells By Ang2-siRNA Plasmid Magnetic Chitosan Nanoparticles Invitro

Objective：Preparation of Ang2-siRNA plasmid magnetic chitosan nanoparticles,and make use of its RNA interference effect to inhibit the expression of angiopoietin2.Explore the efficiency of its inhibition in human malignant melanoma cells. The studywill supply the basis to the further experiment of its targeted interference in nude micetransplanted malignant melanoma angiogenesis and tumor growth in vivo. And it willprovide the experimental evidence for the use of Ang2-siRNA plasmid magneticchitosan nanoparticles in tumor targeted gene therapy.Methods:1. Preparation of Ang2-siRNA plasmid magnetic chitosan nanoparticles.2. Interference of human malignant melanoma cells by Ang2-siRNA plasmidmagnetic chitosan nanoparticles in vitro.3. Observe the expression of red fluorescent protein through inverted fluorescence microscope,then take count of cells and analyze the efficiency of transfection.4. Examine inhibitory efficiency of the expression of Ang2gene in human malignantmelanoma cells by real-time fluorescence quantitative PCR, and get the rate of theinhibition.Results:1. Ang2-siRNA plasmid magnetic chitosan nanoparticles were made successfully.2. The cells transfected successfully show red fluorescence through invertedfluorescence microscope.3. When the mass ratio of Ang2-siRNA plasmid to magnetic chitosan nanoparticles is1:100, the transfection efficiency is61.17%. 4. The results of real-time fluorescence quantitative PCR demonstrated thatAng2-siRNA plasmid magnetic chitosan nanoparticles could inhibit the expression ofAng2genes in human malignant melanoma cells, and the rate of the inhibition was59.56%(p<0.05).Conclusion：Ang2-siRNA plasmid magnetic chitosan nanoparticles can transfecthuman malignant melanoma cells effectively and inhibit the expression of Ang2genes.It will supply the basis and provide experimental evidence for the further experimentof its targeted interference in nude mice transplanted malignant melanomaangiogenesis and tumor growth in vivo.