Genetically Modified Restructuring Of Mage-a3 Tumor Dna Vaccine Construction And Test Research

Cancer is one of the leading cause of death in the world, Advences in molecular biology and our increased understanding of how the immune system function have led to an intense interest in the development of cancer immunological treatment. The cancer specific antigen MAGE-A3 are particularly interesting since their expression shared by different type of tumor, while no in normal tissue. The designation of this study is reconstruction and gene innovation of MAGE-A3 DNA vaccines, to promote effective immunological response and open new therapeutic possibilities.Part I: Objective: Cloning of MAGE-A3 gene. Method: Designed Primer with computer assistant and enlarged MAGE-A3 gene from Mel-526 RNA library by reverse transcription-polymerase chain reaction. Inserted this gene into pGEM-T vector and identify the gene we have cloned. Result: Clone the MAGE-A3 gene successfully; make the cornerstone of farther study.Part II: Objective: To construct MAGE-A3 gene stable expressing animal model of lung cancer. Method: Constructed Eukaryotic expression vector with green fluorescent protein. Transfected inbred strain mouse lung adenocarcinoma cell line LA795 by elect operation and getting stable expressing subclone. Result: Selecting with G418, we got a stable expressing subclone LA795 (MAGE-A1), which remains the characteristic of source LA795 cell, can be transplanted to syngeneic mice T739. This model could do as a preclinic study model for MAGE-A3 targeting irnmunotherapy.Part III: Objective: Construction of MAGE-A3 DNA vaccine vector, innovation this vector and study the immunological effect of these DNA vaccine. Method: Construction of MAGE-A3 DNA vaccine vector with prefabrication method, innovation this vector with a signal peptide from RNATES gene and an IgG Fc segment. Challenged the Immunized T739 mice with the tumor. Result: The gene innovated vaccine can protect syngeneic mice T739 from challenge ofsyngeneic transplanted LA795 (MAGE-A3) tumor.Conclusion: Innovation MAGE-A3 DNA vaccine with a signal peptide and IgG Fc segment can promote effective immunological response and tumor suppression effect. This vaccine strategy may be having some clinic application value.