Anti-tumor Efficacy Of Oncolytic Adenovirus With Fusion Protein Gene 11R-p53

With developing of the society, the incidence of malignant tumors increasingly higher and higher, correspondingly the people have put a lot of manpower and resources in cancer treatment. But in most malignant tumors, many traditionally therapeutic techniques were limited, such as traditional surgery, radiotherapy and chemotherapy and so on. Considered these obstacles, it is very important to optimize cancer therapy. Gene therapy was a kind of new scientific fields for disease therapy recently. Among these gene therapies , many people pay extensive attention on the adenovirus with tumor-specific replication.And now, this kind of virus has been used widely at the research of gene therapy, which can use of the difference metabolic pathways and tissue structure between the tumor and normal tissue cell, only replicate and proliferate in tumor cells, and last, the cancer cells were spallated when reaches a certain level of proliferation, Meanwhile the exogenous genes carried by the plasmid can express for longer. As a vector, proliferative adenovirus have so many advantages,such as: smaller volume, stronger diffuse ability, have the immune response to cancer, the therapeutic genes carried by vector have amplificated many times and so on..P53, as the most important tumor-associated genes, is studied in-depth and extensive, it's basic role is to check the DNA damage in the G1 phase points, and monitor the integrity of the genome. When there is injury ,the P53 gene prevent its DNA replication, so providing more time for DNA repair; if fails ,the cell was triggered apoptosis. other hand, when p53 gene have genic mutation , it's can not control cell proliferation, so this kind cell will turn to cancerous cells. In fact,the P53 gene mutation easilly and loss of function ,the mutation mainly occurre in the highly conserved region,and the P53 gene mutations more than half in human tumors. In addition, P53 gene is also linked inextricably with miRNA, human metabolism, somatic cell reprogramming and so on. Therefore, in this experiment we selece P53 gene as oncolytic adenovirus carrying genes for the treatment of cancer treatment research, and through the synthetic cell-penetrating peptides 11R to enhance its effectiveness. In summary, we have combined oncolytic viruses, therapeutic genes P53 and the cell-penetrating peptides 11R with expression vector to construct the vector SG7605-11R-P53。Then get proliferation-type and adenovirus in 293 cells。We have viewed the capabilities of the virus in various kinds of tumor cells and normal cells ,and the proliferation as well as the cytotoxic ability of the virus in tumor cell .To proviedes a new method and tools for the biological therapy of malignant tumors.After Construction and identification of Vector, and the results in vitro results, we successfully constructed SG7605-11R-P53,,through the Western Blot Detection of the virus in different cells at different time points the expression of the control virus as the SG7605-P53 and Ad-p53, results showed that the expression of proteins for the correct band, and the purpose of Ad-p53 does not appear bands, followed by a wash of anti-P53, by adding one anti-β-Actin and anti-2 to detect the expression ofβ- Actin situation. Secondly, in the MTT experiment, as compared with the control virus, SG7605-11R-P53 can kill tumor cells better.