人肝癌细胞系中肿瘤相关基因MAGE的克隆

目的 克隆人肝癌细胞系HHCC中的肿瘤相关基因MAGE－1的全长cDNA。方法 根据GenBank中MAGE－1基因编码区，设计PCR引物，用RT-PCR手稿,人人肝癌细胞系HHCC中获得MAGE－1全长cDNA，双酶切后连接入质粒pUC19中，转化入大肠杆菌DH5α。挑选阳性克隆提取质粒，进行DNA序列分析，并将测得的核苷酸序列与GenBank中的DNA序列进行BLAST同源性分析。结果 获得MAGE－1全长cDNA,MAGE-6基因463bp片段，以及1个与MAGE－6基因编码区有93％同源性的片段，可能为MAGE家族中的新成员。结论 人肝癌细胞系HHCC可表达多种MAGE基因，可能有效的MAGE基因出现，为研究MAGE基因在HCC中的表达模式及其在HCC免疫治疗中的靶点提供了新的资料。

The cloning of tumor-associated gene MAGE in human hepatocellular cell line

Aim To clone the whole cDNA of tumor associated gene MAGE 1 in human hepatocellar carcinoma cell line HHCC. Methods PCR primer amplifying MAGE 1 gene was designed according to the data in GenBank and the whole cDNA of MAGE 1 was amplified by RT PCR. The PCR product was inserted into the plasmid pUC19 after being digested with restrictive endonucleases. The DNA was sent for sequencing. The result of sequencing and DNA sequence in GenBank was analyzed homologously by BLAST. Results Three clones were obtained, they were: MAGE 1(whole sequence), MAGE 6(463 bp)and a new clone(homology of 93% to MAGE 6, but different from any reported MAGE gene sequence), respectively. Conclusion Several MAGE genes are expressed in human hepatocellular carcinoma cell line HHCC and perhaps there are new MAGE genes. The study provides more date for the researching expression mode of MAGE gene and their targeting site in anticancer immnotherapy.