【作者】 何炜；

【导师】 董子明； 张朝；

【作者基本信息】 郑州大学 ， 生理学， 2005， 硕士

【摘要】 研究背景及目的 黑色素瘤抗原基因家族(MAGE gene family)主要包括四个亚家族：MAGE-A，MAGE-B，MAGE-C和MAGE-D，它们分别位于X染色体的Xq28，Xp21.3，Xq26和Xp11。其中MAGE-A又包含12个基因，从MAGE-1至MAGE-12。MAGE-1基因位于X染色体长臂末端，基因片段长4.5kb，含有3个外显子，其转录的mRNA全长1722bp。MAGE-1编码的肿瘤相关抗原MZ2-E为单链的多肽，包含309个氨基酸残基。近年来，国内外大量文献报道，MAGE-1基因在肝癌、食管癌、胃癌、舌癌、白血病、头颈癌、肺癌、骨肉瘤、膀胱癌、卵巢癌、睾丸癌、神经母细胞瘤等常见肿瘤中都有较高的表达率，而在正常组织、炎症组织及良性肿瘤组织中无该类基因的表达。迄今为止，已发现8个由MAGE-1基因编码的HLA-I类分子限制的CTL表位。因此，利用MAGE-1抗原作为肿瘤疫苗用于肿瘤病人的特异性免疫治疗具有广阔的前景。 本研究利用分子生物学技术，克隆并在体外高效表达MAGE-1基因。构建MAGE-1基因疫苗，并在细胞水平和动物个体水平进行抗肿瘤生物活性方面的研究。为肿瘤的辅助ELISA检测、肿瘤的生物学治疗和瘤苗的研制奠定基础。 材料和方法 从人肝细胞性肝癌(HCC)组织中提取总RNA，通过RT-PCR扩增出MAGE-1基因，酶切鉴定后。将其插入pGEM-T Easy载体，对其进行序列测定并与GenBank已知序列进行同源性分析。再将其克隆至原核表达载体pGEX-4T-1和真核表达更多还原

【Abstract】 Background and objectiveThe MAGE gene family is composed of 23 related genes divided into four clusters located in regions q28(MAGE-A genes), p21.3(MAGE-B genes), q26(MAGE-C genes), and pll(MAGE-D genes) of the X chromosome. The MAGE-A subfamily comprise 12 genes(MAGE-l to MAGE-12) that 60% ~ 98% identity in their coding sequence. The MAGE-1 gene is located in the end of Xq28 and is composed of three exons. The DNA of MAGE-1 is 4.5kb long, while the mRNA of it is 1722bp long. Recently, there are many evidences to show that MAGE-1 gene highly expressed in various type of tumors but silent in normal tissue, inflammation tissue and carcinoid. Up to now, eight MAGE-1 epitopes recognized by cytolytic T Lymphocytes had been found. MAGE-1 antigen is particular interest for antitumor immunotherapy because it is strictly tumor specific and shared by many tumors. The lack of expression of encoding genes in healthy tissues should be ensure strict tumoral specificity of immune response obtained after vaccination of patients with this antigen.The purposes of this study were to develop a MAGE-1 gene vaccine and to observe its effects of anti-tumor in vivo and vitro.更多还原