肝癌组织中三个MAGE家族基因的克隆

目的:从肝癌组织中克隆肿瘤相关抗原基因MAGE-1的全长cDNA,为该基因及其编码抗原应用于肝癌的免疫治疗奠定基础.方法:根据MAGE-1编码区上、下游序列设计一对引物,用RT-PCR方法从肝癌组织中扩增该基因的全长cDNA,酶切产生粘性末端后连接入PUC19质粒.经初步酶切鉴定后,通过DNA序列分析获取所克隆基因片断的核苷酸序列.结果:成功地克隆了MAGE-1基因的全长cDNA,同时还获得一约750bp的MAGE-3基因片段及一与MAGE-6和MAGE-12高度同源的基因的克隆.此与MAGE-6,-12高度同源的基因可能为一未知MAGE家族成员.结论:肝癌组织中表达多种MAGE家族基因,甚至可能存在未知MAGE家族基因的表达,这将为寻找肝癌免疫治疗的攻击靶点开辟新的途径.

Cloning of Three MAGE Genes from Hepatocellular Carcinoma

To clone the full length cDNA of the tumor rejection gene MAGE-1 from hepatocellular carcinoma(HCC) tissues. This MAGE-1 gene and the tumor rejection antigen encoded by it may be useful in subsequent studies aiming at exploring new strategies for the immunotherapy for HCC. Methods: The full length MAGE-1 cDNA was amplified by RT-PCR method using a pair of primers designed according to the encoding sequence of MAGE-1 gene. The PCR products were then digested by restriction endonucleases and inserted into the plasmid PUC19. After primary selection of the recombinants by endonuclease digestion, the sequences of the inserted gene fragments were confirmed by DNA sequence analysis. Results; Using the same pair of primers, we obtained three clones of different MAGE genes, which were a full length MAGE-1 gene, a 750 bp fragment of MAGE-3 gene and a gene highly homologous to MAGE-6 and MAGE-12 but not identical to any reported MAGE genes. Conclusion: These data suggested that some MAGE genes are expressed in heptocellular carcinoma probably including some unknown genes, which might introduce potential new targets for immune attacks.