The Research On Combination Therapy Effect Of Osteosarcoma Cell S-180 By Multiple Gene

Osteosarcoma is a genopathy, one of kind of malignancy. Cell cycle'sderangement is one of pathogenesy in Osteosarcoma. Treatment' method ofosteosarcoma include operation-therapy,chemo-therapy and biotherapy. One ofthem, The Gene-therapy, one of biotherapy, take us the desire of cata-radical cureosteosarcoma. Now, The research about tumor Vaccine,functionality p53,cytokine gene,suicide gene and osteocalcin-gene has received curative effect inosteosarcoma. To study multiple gene Combination Therapy in Osteosarcoma Cell S180, weadopt Fowl pox virus as carrier and admov VP3,HN,IL-18 into the carrier. FPV'sgenome huge and insurmountable, can contain biggish exogenous gene andexpression product can express satisfactory immunogenicity. VP3 gene expressprotein named Apoptin, can selectivity induce tumour cell apoptosis and don'tdamage Normocell. VP3 doesn't depend on P53 and bcl-2 to produce a markedeffect. HN has NA-liveness, can hydrolyze acetylneuraminic acid on cell toexpose surface antigen and to strengthen immunogenicity of cell. IL-18 canstrengthen cytoactive of NK and CTL,can facilitate cell multiplication,canstrengthen toxic action of cell by Fas. We constructed the recombinant fowpox virus vFVHN, vFVVP3,vFVVP3HNIL-18 were screened by using BrdU and identified by RT-PCR,western blot and indirect immunofluorescence. The results of RT-PCR, western blot and indirect immunofluorescenceshowed that foreign gene carried by recombinant fowpox virus could expressefficiently in tumor cells, AO/EB staining, genomic DNA electrophoresis, DCFAstaining to detect the Reactive oxygen species level were used to observe thedeath mode of S180 induced by multiple gene of recombination fowpox virusinfection and which pathway is involved in the cell death process. The resultsshowed that recombination fowpox virus infection caused nucleic condensationand localization, the decline or loss of mitochondrial trans-membrane potential,fragmentation of genomic DNA and the upregulation of p53 in S180 cells, theinfection ultimately induced S180 cell apoptosis and the apoptosis induce byrecombination fowpox virus is mainly through mitochondrial pathway.Furthermore the infection of recombination fowpox virus increased the expressionof MHC-I on the surface of S180 tumor cells and enhanced its immunogenecity.BALB/C mice model bearing S180 hepatoma was construced bytransplanting S180 cells into the right hind limb of the mice and the combinativeantitumor effect on S180 hepatoma of multiple gene was observed through thismodel. The combinative application of multiple gene resulted in the significantantitumor effect than HN gene or VP3 gene alone. According to the above results,we suggested that although the HN gene cannot substitute for the whole virusparticle in antitumor, but it plays an important role in the antigenic augment oftumor cell infected by this virus. The combination of multiple gene has a potentantitumor efficacy.