TNFa在血管内皮细胞ECV304中的靶向表达研究  

Targeted expression of TNF a in ECV304 cells

作　　者：赵显玲[1] 黄百渠[1] 

机构地区：[1]东北师范大学生命科学学院遗传与细胞研究所,吉林长春130024

出　　处：《分子科学学报》2001年第3期145-149,共5页Journal of Molecular Science

基　　金：国家自然科学基金资助项目 ( 39970 384 )

摘　　要：在缺失了 3’LTRU3 区内病毒的启动子 /增强子序列的逆转录病毒载体pLXSNd 中 ,用血管内皮生长因子受体KDR的特异性启动子调控了TNFa在血管内皮细胞ECV30 4中的靶向表达 .将构建的载体 pLXSN - TNFa ,pLXSNd-KDRp-TNFa和空载体 pLXSN用PA317细胞包装后获得重组病毒 ,并用重组病毒分别感染NIH3T3细胞和ECV30 4细胞 .培养物上清的ELISA结果证明 ,KDR启动子指导的TNFa在KDR阳性细胞ECV30 4中的表达量为在KDR阴性细胞NIH3T3中的表达量的 8倍 ;而LTR指导的TNFa在这两种细胞中的表达无明显差异 ,实现了TNFa在血管内皮细胞中的靶向表达 .Results presented in this paper show that TNF a had been specifically expressed in vascular endothelial cells under the control of KDR p in pLXSN whose promoter and enhancer were deleted. Plasmids pLXSN-TNF a and pLXSN d-KDR p-TNF a were constructed and transfected into package cell line PA317 using Lipofectin AMINE method. The supernatant containing the virus was detected with NIH3T3 and was used to transfect NIH3T3 and ECV304 cells. After selection by G418, G418 resistant cell clones were cultured. ELISA analysis of the supernatant collected from tranfected clones showed that TNF-a expression level in ECV304 under the control of KDR p was 8 fold higher than that in NIH3T3, but TNF a expression level under the retroviral promoter had no difference between NIH3T3 and ECV304. The results strongly suggested that TNF a gene in the plasmid pLXSN d-KDR p-TNF a was specifically expressed in ECV304 cells under the control of KDR p. Therefore, this study illustrated the superiority of using targeted expression vectors in cancer gene therapy .

关 键 词：TNFA 靶向表达 血管内皮细胞 ECV304 靶向基因治疗 肿瘤坏死因子 肿瘤 逆转录病毒载体 

分 类 号：R730.5[医药卫生—肿瘤] R394.8[医药卫生—临床医学]