人TACI-Fc融合蛋白对XG6细胞体外生长的调节作用  

The regulatory role of human TACI-Fc fusion protein to XG6 cell line in vitro

作　　者：徐海燕[1] 王泳[1] 於葛华[1] 马泓冰[1] 徐颖[1] 施勤[1] 张学光[1] 

机构地区：[1]苏州大学生物技术研究所

出　　处：《中华微生物学和免疫学杂志》2005年第11期912-916,共5页Chinese Journal of Microbiology and Immunology

基　　金：国家重点基础研究发展计划(2001CB51003);国防科工委重点科研资助项目

摘　　要：目的建立人TACI-Fc基因转染细胞,获得该融合蛋白,研究其对多发性骨髓瘤细胞系XG6的生物学作用。方法利用RT-PCR方法从扁桃体细胞中克隆全长TACI cDNA片段,将该基因的胞外段sTACI与人IgG1Fc段连接到pcDNA真核表达载体上,使两者融合表达。脂质体法将重组质粒pcDNA-sTACI-Fc转入小鼠成纤维细胞系L929,ELISA法测定转染TACI-Fc基因的L929细胞培养上清中TACI2Fc的含量,培养上清经亲和层析柱纯化,获得纯化的融合蛋白TACI-Fc。然后应用台盼蓝染色活细胞计数和3H2TdR掺入法检测其对XG6细胞的生长和增殖的调节作用。结果经RT-PCR和ELISA方法检测表明,成功构建了稳定表达sTACI-Fc融合蛋白的基因转染细胞。基因转染细胞的培养上清经亲和层析柱纯化,得到TACI-Fc融合蛋白。体外检测其生物学活性结果表明,该融合蛋白可有效地阻断Blys对骨髓瘤细胞株XG6的促增殖作用。结论本实验成功地将TACI胞外段与人IgG1Fc段在真核细胞中进行融合表达,表达的蛋白质具有良好的生物学功能,这为进一步对TACI基因进行功能研究奠定了物质基础。Objective To study the effects of TACI-Fc fusion protein on the multiple myeloma cell line XG6 in vitro. Methods The flail length of TACI cDNA was cloned from the tonsil cells by RT-PCR, the extracellular fragment soluble TACI（sTACI） was inserted into expression vector pcDNA, and the human IgGl Fc fragment was ligated with the 3＇ of sTACI fragment. Then the recombinant plasmid pcDNA-sTACI-Fc was transfected into murine fibrocellular cell line L929. The fusion protein TACI-Fc in the suppematants of transfectant L929 culture was analyzed by ELISA. Then the medium was purified by protein G affinity chromatography column for function test. Results The 882 bp eDNA was amplified by RT-PCR. The eukaryotie expression plasmid pcDNA-TACI-Fe was successfully constructed, and the transfectant stably expressed sTACI-Fc was analyzed by RT-PCR and ELISA. The fusion protein can block the prosurvival function of Blys to XG6 in vitro. Conclusion TACI-Fc shows active function vitro.

关 键 词：TACI-Fc融合蛋白 基因转染细胞 增殖 

分 类 号：R392[医药卫生—免疫学]