新型人肿瘤坏死因子基因在血管内皮细胞中的靶向表达  

Endothelial cell targeted expression of human tumor necrosis factorαmutant with higher activity from retroviral vectors

作　　者：石缨[1] 肖畅[2] 赵显玲[3] 王烨[1] 刘丽[1] 朱迅[2] 刘立忠[1] 谢宝树[1] 冷爱军[1] 杨彦[1] 曹颖[1] 

机构地区：[1]空军总医院临床分子生物学中心,北京100036 [2]白求恩医科大学免疫学教研室,吉林长春130021 [3]东北师范大学遗传所,吉林长春130024

出　　处：《免疫学杂志》2001年第1期30-33,共4页Immunological Journal

摘　　要：目的探索一条肿瘤特异性基因治疗的新途径。方法将新型人 TNF- α D11a基因和 KDR特异性启动子插入到 3’L TR缺失 2 99bp的逆转录病毒载体 p L XSN中 ,构建成 p L XSN- D2 99- KDRp- TNF- α D11a重组逆转录病毒载体 ,使目的基因转录受 KDR启动子驱动。通过脂质体介导将该重组载体转染 PA317包装细胞 ,并用病毒上清感染血管内皮细胞和NIH3T3细胞。分别经 EL ISA和 MTT法测定 TNF- α D11a在血管内皮细胞中的表达及其生物学活性。结果成功构建了携带 TNF- α D11a和 KDR启动子的逆转录病毒载体 ,TNF- α D11a在血管内皮细胞中的表达水平及细胞增殖抑制作用均高于NIH3T3细胞。结论 KDR启动子可使外源 TNF-ObjectiveTo develop a new approach in specific gene therapy of tumor Methods TNF α D11a was inser ted downstream of KDR promoter in retroviral vector pLXSN in which 299 bp of 3’ LTR had been deleted The constructed retroviral vectors pLXSN D299 KDRp TNF α D11a were transfected into PA317 packaging cells by lipofectamine reagent Then the recombinant retroviruses were used to infect endothelial cells and NIH3T3 cells Expression and bioactivity of TNF α D11a in culture medium of endothelial cells were identified by ELISA and MTT, respectively Results Retroviral vectors, carrying TNF α D11a and KDR promoter, were constructed successfully Expression level of TNF α D11a in endothelial cells was higher than that in NIH 3T3, and inhibitory effects of TNF α D11a released from endothelial cells on tumor cells prolife ration were also stronger than those from NIH3T3 cells Conclusion Endothelial cell targeted expression of TNF α D11a was regulated by KDR promoter

关 键 词：启动子 TNF-Α 逆转录病毒载体 靶向表达 基因治疗 

分 类 号：R73-3[医药卫生—肿瘤]