黑色素瘤抗原A3基因对肝细胞癌免疫保护作用  

Construction of melanoma antigen A3 expression vector and its immuno-protective effect on hepatocellular carcinoma in vivo

作　　者：张永俊[1] 黄爱民[2] 陈裕庆[2] 高美钦[2] 张文敏[2] 

机构地区：[1] 厦门大学附属中山医院病理科 [2]福建医科大学基础医学院病理学系,福州350004

出　　处：《解剖学杂志》2014年第2期170-174,共5页Chinese Journal of Anatomy

基　　金：福建省自然科学基金（2012J0101）;福建医科大学重大项目（09ZD007）;福建医科大学教授基金（JS08006）

摘　　要：目的：构建小鼠黑色素瘤抗原A3 （MAGE-A3）基因真核表达载体,观察其对小鼠肝细胞癌的免疫保护作用.方法：化学合成小鼠MAGE-A3全长基因,以pcDNA3.1（＋）质粒为载体,构建重组表达载体pcDNA3.1-MAGE-A.将pcDNA3.1（＋）-MAGE-A3载体转染293T细胞,免疫印迹鉴定转染细胞MAGE-A3蛋白的表达.将纯化的重组质粒DNA肌肉注射免疫小鼠,5-Aza-Cdr诱导小鼠肝癌细胞H22表达MAGE-A3基因,观察小鼠成瘤情况与瘤体形态学改变.结果：成功构建pcDNA3.1-MAGE-A3真核表达载体,转染293T细胞后能正常高效表达MAGE-A3蛋白;MAGE-A3重组质粒DNA免疫小鼠后,小鼠成瘤的时间推迟,瘤体生长速度明显减慢,瘤体大小和质量与对照组相比,差异有统计学意义;光镜下显示MAGE-A3重组质粒组肿瘤细胞排列较为稀疏,间质炎症细胞浸润和淋巴细胞增殖更为明显.结论：MAGE-A3基因免疫可有效抑制肝细胞癌成瘤,产生对肝癌的免疫保护作用,提示MAGE-A3基因疫苗可作为肝癌防治手段之一.Objective： To construct murine pcDNA3.1 （＋）-MAGE-A3 eukaryotic expression vector and to observe its immuno- protective effect on hepatocellular carcinoma. Methods： Mice MAGE-A3 gene （C terminal containing 6 × His tags） was synthesized by chemical means. Then the gene was cloned into pcDNA3.1（＋） and amplified in E. coli Dh5a, and then was detected using double digestion identification and gene sequencing. The pcDNA3.1（ ＋）-MAGE-A3 was transfected into 293T cells, and the expression of MAGE-A3 protein was detected by Western-blotting. The mice were immuned for three times, 7 days interval in muscle injection. 5-Aza-Cdr induced the expression of MAGE-A3 in liver cancer cell line H22. Then the MAGE-A3-expressing cell was injected into mice axillary subcutaneous, and the PBS and null vectors injection was used as controls. The tumour growth and its morphous were observed. Results： The recombinant pcDNA3.1 （＋） vector with MAGE-A3 gene was constructed successfully and MAGE-A3 protein was efficiently expressed in 293T cells. The mice were immuned by recombinant vector, followed by MAGE-A3-positive H22 cell implantation. The results showed that the volume and weight of tumor nodules in the recombinant vector group was significantly lower than that of two control groups. The tumor cells arranged sparsely and its periphery had more inflammatory cells in the recombinant vector group. Conclusion .. The MAGE-A3 gene vaccine may efficiently inhibit tumor cell growth, indicating the possibility of gene-based MAGE-3 antigen vaccine as a promising strategy for prevention of MAGE-3-assoiciated cancer.

关 键 词：肝细胞癌 黑色素瘤抗原-A3 真核表达载体 基因疫苗 

分 类 号：R382.31[医药卫生—医学寄生虫学]