舌鳞癌细胞中肿瘤特异性MAGE-A3基因的克隆

Cloning of tumor-specific MAGE-A3 gene from tongue squamous carcinoma cell line Tca8113

目的从人舌鳞癌细胞株Tca8113中扩增出编码MAGE-A3全长抗原蛋白的基因序列，并对其进行测序及鉴定。方法采用逆转录-聚合酶链式反应（RT-PCR）从人舌鳞癌细胞株Tca8113中获取MAGE-A3蛋白编码基因，通过TA克隆后的酶切鉴定以及基因测序鉴定MAGE-A3的正确性。结果（1）RT-PCR产物琼脂糖凝胶电泳结果示，GeneRuler 100bp DNA Ladder的1031bp和900bp之间出现一条明显的目的条带，大小与附加了酶切位点的MAGE-A3目的片段大小相符。（2）双酶切可见大小约3000bp和1000bp的目的条带；单酶切可见大小约4000bp的目的条带，结果与预期相符。（3）经DNA SIS MAX分析软件比对分析，测序结果与Pubmed基因库中查询获得的人MAGE-A3序列一致，MAGE-A3提取成功。结论MAGE-A3的成功提取为进一步研究MAGE-A3抗原蛋白的表达及表达产物诱导特异性抗肿瘤免疫应答提供了实验基础，为抗肿瘤疫苗的研究提供了实验依据。

Objective To clone MAGE-A3 protein encoding gene from tongue squamous carcinoma cell line Tca8113. Methods The MAGE-A3 gene was cloned by RT-PCR from tongue squamous carcinoma Tca8113 cells,and identified by endonuclease and sequencing. Results Agar gel electrophoresis results showed RT-PCR products were between 1031 bp and 900 bp. 3000 bp and 1000 bp target stripes appeared by double enzyme digestion; 4000 bp target stripes appeared by single enzyme digestion. DNA SIS MAX analysis revealed that the cloned gene was consistent with MAGE-A3 gene sequence in Pubmed GENE DATA. Conclusion MAGE-A3 gene has been successfully cloned from tongue squamous carcinoma cell line Tca8113.