基因修饰重组MAGE-A3肿瘤核酸疫苗的构建及试验研究-肿瘤学专业论文.docx

AbstractAbstract Abstract Abstract 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：Objeetive：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 offarther study． Part Ih Objeetive：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(M．AGE·A1)， which remains the characteristic of SOurCe LA795 cell，Can be transplanted to syngeneic mice T739．This model could do as a preclinlc study model for MAGE-A3 targeting immunotherapy． Part III：Objeetive：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 sigrlal peptide from RNATES gene and an IgG Fc segment．Challenged the Immunized T739 mice with the ttullor．Result： The gene innovated vaccine can pmtect syngeneic mice T739 from challenge of l AbS舡actsyngeneic AbS舡act syngeneic transplanted LA795(MA