A Research On The Mechanism Of Improving RAAV Vector’s Gene Expression Efficiency Through MiR-133

As the development of life science, Gene therapy would be one of the bestapproaches for clinic therapy. In comparison to the traditional treatment, gene therapyprovides new method based on gene with strong specificity and persistence. Efficacyof gene expression, accuracy of target and safety are important to vector of genetherapy. Adeno-associated virus (AAV), Lentiviral vector and Adenoviral vectors (Ad)and some retroviral vectors are the common viral vectors of gene therapy. AAV is oneof the most common vector for therapy. It has no pathogenicity, low immunogenicityand the ability to transfer dividing cells as well as non-dividing cells. Although thegenome organization of AAV is simple, its physicochemical properties is very stable.AAV can transfer a wide range of cells and organs. At the same time, differentserotypes of AAV has its own special targets. It is very encouraging that AAV hasbecome one of therapy vectors in Europe.RNA interference (RNAi) is a kind of post-transcriptional gene silencingtechnology and microRNA (miRNA) is one of the small molecules acting as RNAi.miRNA is able to regulate many genes in a signal pathway and even could regulateseveral signal pathways at the same time.Epidermal growth factor receptor (EGFR) regulates the growth, differentiation andproliferation of cells, which is closely related to tumorigenesis. EGFR signalregulates the Ubiquitination of AAV’s capsid and its genome change from singlestrand to double strand. Both of them are the key proceedings during AAV’stransduction.The present study was first time to report that miRNA133(including miRNA133aand miRAN133b) increased the expression efficiency of rAAV in the EGFR-positivecells(such as HeLa and A549cells). miRNA133could also strongly inhibit theproliferation of these cells strongly. miRNA133could target to EGFR, efficiently inthe light of bioinformatics forecast and analysis as well as the experimentalverification, but no influence on the expression of the Lentiviral vector andAdenoviral vectors, suggesting that the specificity of miRNA133should improverAAV expression efficiency. The results of this study revealed the different effects ofmiRNA133and several kinds of medicines on the rAAV expression efficiency, also suggesting that there might and pathways of miRNA133. Our study showed that thecombination of miRNA133with the medicines tested could inhibit the proliferationof tumor cells synergistically, which enhanced the application of miRNA133in thefield of cancer therapy. The specificity of miRNA133between these medicinesindicates many new research points of penetration.miRNA133not only improved rAAV expression efficiency, but also stronglyinhibited the proliferation of tumor cells. The combination of these miRNA133advantages may provide a new approach for cancer gene therapy.