新型人肿瘤坏死因子基因在血管内皮细胞中的靶向表达
신형인종류배사인자기인재혈관내피세포중적파향표체
Endothelial cell-targeted expression of human tumor necrosis factor α mutant with higher activity from retroviral vectors
  • 万方数据
    免疫学杂志 면역학잡지
    IMMUNOLOGICAL JOURNAL
    2001年 1期 30-33 ,共4页

    石缨%肖畅%赵显玲%王烨%刘丽%朱迅%刘立忠%谢宝树%冷爱军%杨彦%曹颖

    석영%초창%조현령%왕엽%류려%주신%류립충%사보수%랭애군%양언%조영

    启动子%TNF-α%逆转录病毒载体%靶向表达%基因治疗 啟動子%TNF-α%逆轉錄病毒載體%靶嚮錶達%基因治療
    계동자%TNF-α%역전록병독재체%파향표체%기인치료
    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-αD11a基因和KDR特异性启动子插入到3’LTR缺失299bp的逆转录病毒载体pLXSN中，构建成pLXSN-D299-KDRp-TNF-αD11a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ELISA和MTT法测定TNF-αD11a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-αD11a和KDR启动子的逆转录病毒载体，TNF-αD11a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-αD11a在血管内皮细胞中特异性表达。
    목적 탐색일조종류특이성기인치료적신도경。방법 장신형인TNF-αD11a기인화KDR특이성계동자삽입도3’LTR결실299bp적역전록병독재체pLXSN중，구건성pLXSN-D299-KDRp-TNF-αD11a중조역전록병독재체，사목적기인전록수KDR계동자구동。통과지질체개도장해중조재체전염PA317포장세포，병용병독상청감염혈관내피세포화NIH3T3세포。분별경ELISA화MTT법측정TNF-αD11a재혈관내피세포중적표체급기생물학활성。결과 성공구건료휴대TNF-αD11a화KDR계동자적역전록병독재체，TNF-αD11a재혈관내피세포중적표체수평급세포증식억제작용균고우NIH3T3세포。결론 KDR계동자가사외원TNF-αD11a재혈관내피세포중특이성표체。
    Objective To develop a new approach in specific gene therapy oftumor.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.
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