文章摘要
Lu Xu,Jin Zhu,Zhenning Qiu,Yuhua Li,Zhenqin Feng.[J].南京医科大学学报,2007,(5):287~292
Cloning of human melanoma antigen MAGE-A9 and its expression in hepatocellular carcinomas
  
DOI:10.7655
中文关键词: 
英文关键词: melanoma antigen  MAGE-A9  RT-PCR  gene expression  immunohistochemistry
基金项目:
作者单位
Lu Xu Department of Pathology, Nanjing Medical University, Nanjing 210029, Jiangsu Province, China 
Jin Zhu Institute of Molecular Research, Nanjing Medical University, Nanjing 210029, Jiangsu Province, China 
Zhenning Qiu Institute of Molecular Research, Nanjing Medical University, Nanjing 210029, Jiangsu Province, China 
Yuhua Li Department of Pathology, Nanjing Medical University, Nanjing 210029, Jiangsu Province, China 
Zhenqin Feng Department of Pathology, Nanjing Medical University, Nanjing 210029, Jiangsu Province, China 
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中文摘要:
      
英文摘要:
      Objective:To express the melanoma associated gene MAGE-A9 recombinant protein, obtain the anti-MAGE-A9 monoclonal antibody and to examine the expression of MAGE-A9 in hapatocellular carcinoma specimens. Methods:MAGE-A9 cDNA was cloned from human hepatocellular carcinoma tissue by using RT-PCR, and then subcloned into the plasmid pMD18-T. After sequencing, the MAGE-A9 was cloned into the prokaryotic expression vector pBAD/gIII to construct the recombinant expression vector pBAD/gⅢ-MAGE-A9, and was transformed into E. coli TOP10. The recombinant MAGE-A9 protein was expressed under induction of L-Arabinose, and was purified through Hitrap column. The anti-MAGE-A9 monoclonal antibody was generated. The expression of MAGE-A9 in hepatocellular carcinoma specimens was examined through ABC assay. Results:The cDNA sequence of the cloned MAGE-A9 gene was consistent with the reported sequence. By affinity column and SDS-PAGE, the purified MAGE-A9 fusion protein displayed a band of Mr 35,000, and subsequently the anti-MAGE-A9 monoclonal antibody was obtained. We found that MAGE-A9 expressed in the cytoplast of positive cells and MAGE-A9 antigen was detected in 8 cases out of 39 (21%) hepatocellular carcinoma specimens. Conclusion:MAGE-A9 antigen was expressed in a fair proportion of hepatocellular carcinoma specimens, these patients might be suitable candidates for immune involving antigen, encoded by the MAGE-A9 gene.
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