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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方法 ,从人肝癌细胞系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 4 6 3bp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 ,可能有新的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。
Objective To clone full length cDNA of tumor associated gene MAGE 1 in human hepatoma cell line HHCC. Methods According to the coding region of MAGE 1 gene in GenBank, PCR primers were designed. The full length cDNA of MAGE 1 was obtained from human hepatocellular carcinoma cell line HHCC by RT PCR. The double digested DNA was ligated into plasmid pUC19 and transformed into E. coli DH5α. The positive clones were selected for plasmid extraction, DNA sequence analysis and nucleotide sequence analysis and the GenBank DNA sequence BLAST homology analysis. Results The MAGE 1 full-length cDNA, the MAGE 6 gene 46bp fragment, and a fragment of 93% homology with the MAGE 6 gene coding region were obtained, which may be a new member of the MAGE family. Conclusion Many human hepatocellular carcinoma cell lines HHCC express a variety of MAGE genes, and some new MAGE genes may appear. It provides new information for studying the expression pattern of MAGE gene in HCC and its target in HCC immunotherapy.