The Influence Of Recombinate Adenovirus Vector To Salivary Gland Cell And The Synthesis Of Magnetic Nano Fe₃O₄ Gene Vector

Salivary tumors are frequent tumor in oromaxillo-facial region.Adenoid cystic carcinoma (ACC) is a common salivary malignant tumor, which infiltrates facial nerve and metastasis earlier and often leads to obvious maxillo-facial malformation after operation. It also reoccurs easily and is not sensitive to radio-therapy or chemo-therapy, so has a deep effect to patient's life.It is very important to approach it's reason and find new effective therapy for it.Gene therapy have develop more than 20 years since 1980s.In 1990,NIH used ADA gene cure a 4 years old girl who had severe immune deficiency for ADA gene defect.In 1994, Shillitoe advocate that use gene therapy to treat oral tumors.Gene therapy have marked achievement during last 10years.It has played more and more therapy function for hereditary disease,metabolic disease and malignant tumor.But when deliveried in the aqueous phase,nucleic acid are easy to be hydrolysis by nuclease,and because nucleic acid have a lot of negative charge,it can't be swallowed by cell easily.For these reason ,we must solve the key problem that find high performance,safe and tissue-specific gene vector.There are two kinds of gene vectors, viral vectors and nonviral vectors.Viral vectors are the most effective up to now such as etroviral vectors, adenoviral vectors, adeno-associated viral vecto rs,herps simplex viral vectors, lentiviral vectors et al.But this kind vector exist side effect and immune reaction to confine it's widespread application.So people began to pay attention to the safe nonviral vectors,especially the development of nano material.Nanoparticles which size between 0.1-100nm ,also called ultramicroparticle,are smaller than common cell,so it's possible to be vector to enter cell.Nanoparticle have a lot of physical and chemical character for example large specific surface,surface atomic number,surface energy and surface tension rapid increase with the particle diameter decrease.All these character lead to specific heat,magnetic,photaesthesia character so we can control nano vector target deliver gene by electricity,magnetic and light.We also can modify nanoparticle's surface structure and condition to raise surface activity,to improve nanoparticle's compatibility and to form new physical , chemical and mechanism character and new function.Magnetic nanoparticle is a important kind of nanoparticle.It has superparamagntism,so can easily phase separation and localization by magnetic field. So the Magnetofection by the Magnetic nanoparticle may be a valid tumor therapy method.Nanotechnology,biotechnology and bioinformatics have been generally accepted as motive power for medical science development.Hitherto ,there are great stride in vivo and in vitro research.Maybe in recent future, nanobiotechnology will become more effective therapy for genopathy than chemo-therapy, radio-therapy and surgical treatmentThe salivary glands of mammalian (parotid and submandibular gland) are more excellent target of gene therapy due to their anatomy characteristic than other internal organ.It's location superficial so easily observe the transfection effect.We believe salivary tumors gene therapy have potential application perspective with the research development.In this study, we aim directly at salivary to test the safe problem of recombinate adenovirus vector and prepare the Fe₃O₄@PDMA-pACTERT-TRAIL magnetic nano gene vector.We try to develop new magnetic nano gene vector to target therapy salivary tumor .The field of this research is following: 1 .We used collagenase and trypsinization degested method culture normal Wistar rat submaxillary salivary gland cells.Then transfected the salivary cells with recombinate adenovirus vector which express enhanced green fluorescent protein with CMV as promoter.The results show when we used higher MOI( more than 200 particle/cell ) recombinate adenovirus vector can inhibited the proliferation of the salivary cells and influented salivary cells secrete amylase. So it is toxiferous for salivary cells.In the other hand,the result also indicate that used proper MOI ,the adenovirus vector expressed EGFP can be used to mark salivary cells for trace labeling and research submaxillary cells's biological behaviour. 2.We used ARTP reaction prepare Fe₃O₄@PDMA magnetic nano gene vector which surface contain positive charge.Then Fe₃O₄@PDMA combinate with plasmid by electrostatic interation.We used electrophoresis et al methods to detect the complex.The result show they successful combinated.Our research achievements provide a foundation for us to investigate the target transgene ability of magnetic nanoparticle in vivo and in salivary tumors, as well as theory basis and practice experience for us to conduct gene therapy clinically with high efficiency, targeting effect and no trauma, which will develop a new way to treat salivary tumors .