重组腺病毒Ad5/F11p-GFP对不同肿瘤细胞系感染效率的比较研究

摘要

目的以 Ad5-GFP 及 Ad5/F35-GFP 为对照，评价Ad5/F11p-GFP 对不同肿瘤细胞系的感染效率。n　　方法制备并纯化 Ad5-GFP、Ad5/F11p-GFP 及 Ad5/F35-GFP三种重组腺病毒，以不同的感染复数（MOI）感染乳腺癌细胞系 SKBR-3、MCF-7及肾癌细胞系786O。48 h 后，荧光显微镜观察 GFP 的表达，流式细胞仪检测感染效率及不同细胞表面柯萨奇腺病毒受体（CAR）、CD46和桥粒芯糖蛋白质2的表达。n　　结果 Ad5/F35-GFP 对 SKBR-3、MCF-7及 786O 均具有很高的感染效率，且其在较低的 MOI 时就具有较高的感染效率；Ad5/F11p-GFP 对 CD46高表达的 MCF-7及 786O具有较高的感染效率，5 MOI 感染即可达到60%以上的感染效率；而 Ad5-GFP 仅对 CAR 高表达的细胞系786O具有较高的感染效率，且感染效率随着 MOI 的增加而升高。Ad5/F11p-GFP 和 Ad5/F35-GFP 对三种肿瘤细胞系的感染效率明显高于 Ad5-GFP。n　　结论对5型腺病毒进行纤维顶球的改造，可增强对肿瘤细胞的感染效率。%Objective To evaluate the infection efficiency of recombinant adenovirus Ad5/F11p-GFP in different tumor cell lines by using Ad5-GFP and Ad5/F35-GFP as control adenovirus. n Methods Three kinds of recombinant adenovirus Ad5-GFP, Ad5/F11p-GFP and Ad5/F35-GFP were prepared and purified to infect SKBR-3, MCF-7 and 786O cell lines with different multiplicity of infection (MOI). After 48 hours, the expression of green fluorescent protein was detected by fluorescence microscope. The expressions of coxsackie virus and adenovirus receptor (CAR), CD46 and Desmoglein 2 on SKBR-3, MCF-7 and 786O cell lines were detected by flow cytometry. n Results The infectious effects of Ad5/F35-GFP in SKBR-3, MCF-7 and 786O cells were higher at lower MOI. The infectious effects of Ad5/F11p-GFP in MCF-7 and 786O, which had high expression of CD46, were higher. It could infect more than 60%of cells at 5 MOI. But Ad5-GFP only had high infection efficiency in 786O cell lines which had higher expression of CAR, and the infection efficiency was improved with increasing of MOI. Ad5/F11p-GFP and Ad5/F35-GFP had higher infection efficiency in SKBR-3, MCF-7 and 786O cells than Ad5-GFP did. n Conclusion Fiber-modification to Ad5 vector which is derived from chimeric adenovector containing Ad11p fiber might posses high infection efficiency in tumor cell lines.