Study On The Treatment Of Osteosarcoma With Superparamagnetic Liposome Loaded With Survivin Gene

Objective:Superparamagnetic liposomes were prepared as gene vectors and vehicle conditions and biosafety were evaluated.Optimize in vitro transfection conditions and improve magnetic transfection efficiency.To investigate the effects of magnetic liposomes carrying Survivin interference gene on the biological behavior of osteosarcoma MG-63 cells.Methods:1.Fe₃O₄ nanoparticles were prepared by chemical coprecipitation method,and they were made into magnetic liposomes with Lip2000.The plasmid binding ability was observed by agarose gel electrophoresis.A Zeta potential analyzer and a vibrating sample magnetometer?VSM?are used to detect the zeta potential and magnetic properties of the carrier.Transmission electron microscopy?TEM?was used to observe the ability of transfected cells.Cell scratching and MTT assays were used to assess the cytotoxicity of magnetic liposomes.2.Inverted fluorescence microscopy was used to observe the expression of GFP green fluorescence after transfection of human renal epithelial cells?HEK-293?under different magnetic field action time,thereby optimizing transfection conditions and transfecting osteosarcoma cells?MG-63,143b?.)Ability.Flow cytometry was used to detect the intensity of GFP expression after transfection and to evaluate transfection efficiency.3.Construction of Survivin interference plasmid,CTL luminescence method,CCK8 method to detect the effect of magnetic liposome-loaded interfering gene on the activity and proliferation of MG-63 cells;the effect of Tanswell on cell migration;PI-Hoechst kit detection on MG63 The effect of apoptosis.Results:1.Gel electrophoresis showed that the magnetic liposome containing the magnetic nanoparticle dilution factor of 50X was constructed to bind to the plasmid.Potentiometric Determination The zeta potential of the magnetic liposome was 27.94 mV with a positive charge.The hysteresis loop shows that the magnetic liposome has good magnetic responsivenessandsuperparamagnetism.Transmissionelectron microscopy revealed that the magnetic liposomes entered the cells through the membrane depression and were distributed in the cytoplasm in large amounts.Both the cell scratch test and the MTT assay showed that the prepared magnetic liposome had no significant cytotoxicity.2.The cell transfection experiment showed that the transfection efficiency reached the highest under the condition of external magnetic field for 15 minutes.Under in vitro conditions,the survivin interference gene carries a significant inhibitory effect on the proliferation and migration of osteosarcoma MG63 through efficient magnetic transfection efficiency,and can promote its apoptosis.Conclusion:The constructed magnetic liposome is superparamagnetic and positively charged.It can be used as a good gene carrier,greatly improve the transfection efficiency of the gene,and can efficiently transfer the target gene into the target cell,thereby exerting therapeutic effects.This study showed a good inhibitory effect on osteosarcoma MG-63 cells by carrying the Survivin interference gene in vitro,providing new methods and ideas for gene therapy of osteosarcoma,and also showing a new way for the treatment of other diseases.