Antitumor Effect Of Harboring IL-24 And IFN-β Gene Mediated By Targeted Oncolytic Adenovirus In Cancer

In the context of precision medicine and personalized treatment gradually recognized by the public.Academician of Chinese academy of sciences xin-yuan Liu created Cancer Targeting Gene-Viro Therapy（CTGVT）as a new strategy in 2001.This is a new treatment method which combining gene therapy with virus therapy.By inserting the therapeutic gene into the virus,with targeting into the tumor,the replication of the virus and the amplification of the corresponding gene can inhibit the growth of tumor cells.In this study,we use the CTGVT strategy to insert foreign genes to TK area to enhance the targeted effect of cancer cell killing.Virus targets to tumor cells with a large number of copies while the therapeutic gene IL-24 and IFN-βreplicate at the same time.Therefore,the anticancer effect is greatly improved,and then the treatment of tumor is achieved.And we use the tumor specific Survivin promoter,which is key element for adenovirus replication,and delete the 55bp DNA sequence located on E1Bregion to construct a novel oncolytic adenovirus named Onco^Ad-surp-E1A-E1B（Δ55）.Followed by the technique of gene recombination which cloning IL-24 and IFN-βgene into oncolytic adenovirus vector Onco^Ad-surp-E1A-E1B（Δ55）,the new recombinant oncolytic adenovirus of dual gene Onco^Ad-（IL-24）-surp-E1A-E1B（Δ55）-（IFN-β）is produced.As control,we also used new adenovirus structure system to construct empty virus and single gene virus Onco^Ad-surp-E1A-E1B（Δ55）,Onco^Ad-（IL-24）-surp-E1A-E1B（Δ55）and Onco^Ad-surp-E1A-E1B（Δ55）-（IFN-β）.After the successful identification process,titer matched experimental sample are obtained by purified of CsCl gradient centrifugation.With comparison of cell experiments between four viral groups and non viral control groups,the impact to hepatoma carcinoma cell with morphological changes,apoptotic body formation,cell viability,apoptosis and the expression of apoptosis signal transduction related protein from double gene targeting oncolytic adenovirus are tested by fluorescence microscope,Hochest33342 staining,MTT assay,AnnexinV/PI flow cytometry and Western blotting et al.The results showed that the adenovirus carrying the target gene was effective in killing cancer cells.Through the relevant experiments and lots of articles for the query,we speculate that,after infecting hepatoma cell by the adenovirus carrying the target gene,hepatoma cells downstream inhibiting apoptosis related gene activation level is reduced and the endoplasmic reticulum stress continuous eventually trigger caspase cascade to the apoptosis pathway.Our study is the exploratory research for adenovirus vector,IL-24 and IFN-βgene in the application of gene therapy provides reference,as well as provides a theoretical conception and framework for precision personalized medicine and cancer therapy.