Study On The Preparation Of Co-transporter Assembled By CDPNTs Ca Rry SiRNA And The Anti-tumor Activities

RNA interference(RNA interference, RNAi) technology is a double-stranded RNA(ds RNA) that can specifically and effectively promote the degradation of homologous m RNA, thereby blocking expression of the target gene phenomenon. The RNAi mechanism is Widespread in biological cells, and because it can be quickly and easily block the expression of target gene. In recent years, it has become one of the effective means of gene therapy. Among them, the use of RNAi technology block the process of tumor cell development in specific signaling pathways. It can be efficiently and easily promote the apoptosis of tumor cell, is currently one of hot topic in anti-tumor therapy.Currently, RNAi technology in clinical application process bottlenecks facing the main problem is the effective delivery of si RNA. Excellent si RNA delivery vehicle should have the following elements:(1)Easy si RNA binding, and can protect the si RNA being ribonuclease(Ribonuclease, RNase) degradation, while carrier and si RNA complexes can not affect to play a role that si RNA enter the cells;(2)High transfection efficiency of the support material;(3)On the specific tissue or specific cells targeting can improve the specific tissue or cell transfection efficiency, thereby enhancing the efficiency of specific tissues or RNAi of cells;(4)Material having a low toxicity, are unlikely to cause the body’s immune response, biocompatibility, and easy in vivo degradation.By now known si RNA delivery vehicle comparison, we found that the cationic polypeptide carrier is one of the ideal carrier material. Having ease of synthesis, adjustable size, structure of easy modification, targeting ligand may be modified so as to have targeted positioning function, and the material properties of stability, low cytotoxicity, susceptible to metabolic advantages in an organism.In this paper, we mainly study the cytotoxicity of CDPNTs carrier material, si RNA binding efficiency, transfection efficiency, the m RNA interference effects of survivin gene, induced apoptosis of SK-OV-3 cells. Test results show low cytotoxicity of CDPNTs material. When CDPNTs concentrations were 10 and 100 μg / m L, SK-OV-3 cells 48 hours survival rates were 88.241 ± 1.739% and 83.102 ± 5.428%. The positive control group Lipofectamine2000(5μg / m L) cell viability is 80.440 ± 5.842%, this shows that a concentration of 10-100 μg / m L of CDPNTs cytotoxic were lower than the Lipofectamine2000; We demonstrate that CDONTs can be combined with si RNA stably by agarose electrophoresis. The si RNA enter into the cell with FAM-labeled deliveried by CDONTs. Observed by fluorescence microscopy, the FAM-si RNA is concentrated in the cytoplasmic matrix. The si RNA deliveried by CDONTs may play a normal RNAi role in the cell. After 48 hours we use the CDPNTs delivery si RNA with inhibiting apoptosis survivin gene, the expression of survivin gene m RNA was significantly inhibited in the cell and apoptotic cells were found.