The Study Of Recombined Vaccinia Virus Carrying ING4 And GMCSF In Targeting Gene Therapy

Ⅰ.In recent years, cancer has become one of the most important diseases threatening human beings. Because of limitation by a variety of reasons in traditional therapy for treatment of cancer, the effect is not ideal and the adverse effect is obvious. With the development of medical science and technology, the continuous exploration and update of cancer treatment methods, the biological therapy of cancer has brought the dawn to the patients. Combination of gene therapyand viral therapy, the cancer targeted gene-viral therapy strategy was presented by academician Liu Xinyuan.This strategy proposed new targets for the selection of viral vectors and therapeutic gene. Vaccinia virus with some advantages of high safety, quick replication and large packaging volumes can be an ideal vector for gene therapy. Previous studies showed that the ING4 gene is widely involved in the process of tumor development, resisted cancer includes inhibiting the activity of HIF and enhancing the activity of P53 gene, which can promote tumor cell death. GM-CSF is a granulocyte macrophage colony stimulating factor, which can activate the body’s immunity to enhance the anti-tumor effect.In this study, we applied vaccinia virus as a carrier to carry two therapeutic gene ING4 or GM-CSF and packaged oncolytic vaccinia virus endowed with high safety and significant killing effect. Recombinational vaccinia was ultilized to destruct lung carcinoma, breast cancer, liver cancer, colon cancer, pancreatic cancer, brain glioma, and then, the observation of cell morphological changes, analysis of crystal violet staining, inducing apoptosis effect using Hoechst staining, detection of the cell survival rate with MTT assay and the expression of related proteins via Western blot were performed.The findings indicated that the oncolytic vaccinia virus OncoPox-ING4 had the most significant effect on inducing cancer cell apoptosis, and also verifiedthat tumor growth could be inhibited by ING4 in a certain link of NF-KB signaling pathway. Therefore, it is concluded that the OncoPox-ING4 has a broad spectrum of killing cancer cells and remarkable anti-cancer effect, which proves the rationality of the "targeted gene virus therapy strategy" and this will provide a new method for the treatment of cancer.Ⅱ.Virotherapy on the basis of oncolytic adenovirus is a novel approach for human cancer therapeutics. We aim to investigate whether it will synergistically reinforce their anti-pancreatic cancer activities when the combined use of ZD55 TRAIL-IETD-Smac and SNS-032 was performed. The experiments in vitro showed that the CDK inhibitor SNS-032 enhances ZD55-TRAIL-IETD-Smac-induced apoptosis and causes remarkable pancreatic cancer cell death. Western blot assays suggested that the SNS-032 could intensify ZD55-TRAIL-IETD-Smac-induced apoptosis of pancreatic cancer by affecting anti-apoptotic signaling elements, such as CDK-2, CDK-9, Mcl-1 and XIAP. Besides, animal experiment further confirmed that combination of SNS-032 and ZD55-TRAIL-IETD-Smac achieved significant inhibition of Bx PC-3 pancreatic tumor xenografted growth. In summary, we have demonstrated that SNS-032 can sensitize human pancreatic cancer cells to ZD55-TRAIL-IETD-Smac-induced cell death in vitro and in vivo. These findings indicate that the combined treatment with SNS-032 and ZD55-TRAIL-IETD-Smac could represent a rational approach for antipancreatic cancer therapy.