Study Of Transfect Dendritic Cell Vaccine Of Recombinant Plasmid PchIL-18-MAGE In Liver Cancer Cell In Vitro

Objective:To observe lethal effect on liver cancer cells by the DCs transfected withrecombinant plasmid pchIL-18-MAGE.Liver cancer is one of the mostprevalent malignant diseases worldwide. Many locoregional therapeuticapproaches, including surgical resection, radiofrequency ablation (RFA),percutaneous ethanol injection(PEI), and transcatheter hepatic arterialchemoembolization (TACE) have been applied in the search for curativetreatments for HCC. Unfortunately, there is still a poor prognosis and highrecurrence rate of liver cancer. Dendritic cells (DCs) are the most potentantigen-presenting cells. The immune functions of DCs in a number of tumorshave been identifid. DC vaccination is one of the new modalities ofimmunotherapy in cancer that posses less frequency of complications, minimalinvasive procedures and very good tolerability by most of the patients. It playsan important role in tumor biological therapy. Melanoma antigen-encodinggene-1(MAGE-1) is one of the members of MAGE family, it is not expressedin normal tissues (except testis and placenta), but in melanoma and othermalignant tumors, especially with a high expression in liver cancer; IL-18is acytokine that has a variety of biological activities, which can activate NK cells, induce NK to secrete IFN-γ. So IL-18can be used as a immune adjuvant toenhance the immune effect of vaccine. This study is to observe the lethal effecton liver cancer cells by the DCs transfected with recombinant plasmidpchIL-18-MAGE.Methods:Cultivate dendritic cells in vitro, recombinant plasmid pchIL-18-MAGEwas constructed and used to transfect DCs．Expression of pchIL-18-MAGE onthe transfected DCs were detected by RT-PCR and Western blot, and thephenotype of DCs were analyzed by flow cytometry. Stimulated lymphocytes bytransfected DC as effector cells, and these by non-transfected DC as control,Detect killing effect of target cells in vitro. IFN-γ in the culture supematant ofthe transfected DCs were detected by ABC-ELISA.Results:pchIL-18-MAGE transfected DCs expressed higher level of CD83、CD1a、CD86、 CD80、 HLA-DR， and expressed as a mature DC phenotype.pchIL-18-MAGE group’s killing activity was the highest （P<0.05）．Conclusion:The DC vaccine pchlL-18-MAGE can induce better killing effect comparedwith other experimental groups．PchIL-18-MAGE transfected DCs have aobvious killing effect to MAGE+liver cancer cells.