The Experimentation Of Ang2-siRNA Plasmid Chitosan Magnetic Nanoparticles Inhibit The Growth Of Portability Tumor Of Malignant Melanoma In Nude Mice

ObjectiveWe build the malignant melanoma nude mice model and inject the Ang2-siRNA plasmid chitosan magnetic nanoparticles into mice.Intervening the portability tumor of malignant melanoma,to observe the transplanted tumor angiogenesis and tumor growth of malignant melanoma in nude mice,which lay the foundation for the further study of targeted inhibition in angiogenesis and tumor growth of malignant melanoma.MethodsWe culture and passage malignant melanoma A375 cells,malignant melanoma cells which were in logarithmic phase would be made cell suspension,and adjust the cell density to 5×107 cells/ml.Inoculating cell suspension in the right axillary subcutaneous of 20 male nude mice.Each male nude will be injected 100 ul cell suspension.After 3 days,malignant melanoma nude mice will be divided into 2 groups at random,including the control group and the experimental group.The nude mice in control group will be injected 0.4ml normal saline without magnetic.The nude mice in experimental group will be injected 0.4ml Ang2-siRNA plasmid chitosan magnetic nanoparticles and under the 4000 GS magnetic field cling right axillary subcutaneous.After 60 mins,magnetic field will be removed.To observe and record the proliferation and volume of transplanted tumor,analyze the volume difference of transplanted tumors between the groups,and make the tumor growth curve.With real-time fluorescence quantitative RT-PCR method to detect relative Ang2 gene expression in transplanted tumor volume,Ang2 gene expression levels between groups in terms of statistical analysis of relationship between degree of difference and tumor proliferation.Using Western blot detection Ang2-siRNA plasmid chitosan magnetic nanoparticles in nude mice on the expression of VEGF,Akt and PI3 K proteins.With immune histochemical method to detect transplanted tumor microvascular density,differences between tow groups in terms of statistical analysis and its relationship with the degree of tumor proliferation.Malignant melanoma with Tunel staining method to detect nude mice cell apoptosis rate.ResultsThe targeted study of the Ang2-siRNA plasmid chitosan magnetic nanoparticles have confirmed that the particles can move to the targeted tissues under an additional magnetic.Human malignant melanoma transplanted tumor nude mice model was to establish well.Tumor volumes of experimental group were significantly smaller than the control group;The Ang2 mRNA expression was significant lower to the control group;The microvessel density was notably lower to the control group;The apoptosis rate of transplanted malignant melanoma was notably lower to the control group;Ang2-si RNA plasmid chitosan magnetic nanoparticles in nude mice about the expression of VEGF,Akt and PI3 K proteins have no significant differences between the control group and the experimental group.ConclusionsThe study of the Ang2-siRNA plasmid chitosan magnetic nanoparticles have confirmed that the particles have the characteristics of targeted drug delivery systerm,which can be a reliable and efficient targeted drug deliery carrier and have a great application in the tumor target therapy.The Ang2-siRNA plasmid chitosan magnetic nanoparticles can effectively reduce the level of gene expression of Ang2 in human malignant melanoma transplanted tumors;thereby inhibit the angiogenesis and tumor growth of the tumor.