Construction And Identification Of IASPP Antisense Gene Magnetic Nanoparticies

Background and Objective:Pancreatic cancer is a maligant tumor which highly related with p53.This anti-oncogene is related to apoptosis stimulating protein of p53 which discovered recently concluding ASPP1,ASPP2 and inhibitory member of the ASPP family (iASPP).The iASPP is an key oncoprotein to cause the tumor,which related inhibits wild type p53-dependent apoptosis and objective gene transcription.In the gene therapy,the iASPP is a target of molecule because the iASPP is a profilin of conserved in p53 evolutionarily.To compose iASPP antisense gene and to study its stability and the biological effects to pancreatic cancer cells.To construct iASPP antisense gene magnetic nanoparticies and evaluate magnetic nanoparticies'protective effect to antisense gene,to evaluate the iASPP antisense gene magnetic nanoparticies could move direction in magnetic field.Materials and Methods:Fristly,using the technology of genetic engineering to design and synthesis of iASPP antisense gene,and by culturing pancreatic cancer cells of SW 1990 at the same time to study the stability of antisense gene and the effect of proliferation and apoptosis of SW1990.Secondly,by coupling the antisense gene and magnetic nanoparticles to construct iASPP antisense gene magnetic nanoparticies.Finally, to prove if the magnetic nanoparticles can protect iASPP antisense gene against nuclease digestion and to test directional movement in the magnetic field.Results:The iASPP antisense gene of manmade is stability in the culture medium during 5 days,which can inhabit the proliferation of SW1990 cells significantly,the highest inhibitive rate can reach to 91.16%.After treatment with the iASPP antisense gene the cells in S phase were decreased and the cells G0/G1 phase increased remarkably.It showed that the cells in G0/G1 phase were blocked and S phase cells decreased,which indicate that reducing the DNA replication and leading to the apoptosis.This mechanismis is likely to P53.We succeedly synthesized iron oxide nanoparticles of binding iASPP antisense gene in the experiment which showed that nanoparticles has the satisfactory potential to adsorb DNA and nanoparticles can effectively protect DNA from DNA polâ… digestion.The shape of magnetic nanoparticies is irregular,with iron nanoparticles in the center and the diameter less than 100 nm. Another experiment showed the directional movement of the iASPP antisense gene nanoparticles under magnetic field.Conclusions:1.The synthetic iASPP antisense gene is stability in the SW1990 culture fluid of pancreatic cancer cell by thio-phosphotylation modification.It can alter the cell growth and inhibite the cell proliferation,so it can help to therapying the pancreatic cancer.2.The iASPP antisense gene can couple with magnetic nanoparticies in a certain quality rate by electrostatic force.The composite can against the digestive enzyme and make directional movement in the magnetic field.