Gene Expression of B16F10 Induced by Treatment with 5-aza-2'-deoxycytidine / 대한이비인후과학회지

ABSTRACT

BACKGROUND AND OBJECTIVES: The goal of tumor vaccine is to activate immune system specifically against proteins expressed by a tumor, and many types of vaccines such as gene modified vaccines have been developed to increase immunogenicity of vaccine. We studied to determine whether or not 5-aza-2'-deoxycytidine (ADC) can increase the immunogenicity of B16F10 melanoma cell. MATERIALS AND METHOD: B16F10 cell was treated with ADC for the induction of DNA demethylation. An ADC treated B16F10 melanoma cell was analyzed first using the reverse transcriptase-polymerase chain reaction (RT-PCR) technique to evaluate the gene expression of tumor antigen (MAGE-2, MAGE-5) and immunity-enhancing cytokines (GM-CSF, IL-12), and then by flow cytometry to evaluate the expression of MHC and B7 that are responsible for antigen expression and T cell activation on B16F10 cell surface. In order to evaluate vaccination effect of ADC-treated B16F10 vaccine, each mouse group were injected with PBS, ADC, B16F10 vaccine or B16F10-ADC vaccine and they were also challenged with live B16F10 cell 7 days after vaccination. On the 20th day after live B16F10 cell challenge, the tumor mass size and the mouse survival period were determined. RESULTS: ADC treatment for B16F10 melanoma cell increased expression of MHC and B7. ADC treatment also increased gene expression of MAGE-2, MAGE-5, GM-CSF and IL-12. The growth of tumor mass was decreased and the mouse survival period was elongated in B16F10-ADC vaccine immunized group. CONCLUSION: ADC treatment may increase immunogenicity of B16F10 cell, and B16F10-ADC vaccine immunization can induce anti-cancer immunity in vivo.