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。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。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。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. 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 786 O 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 786 O cell lines were detected by flow cytometry. Results The infectious effects of Ad5/F35-GFP in SKBR-3, MCF-7 and 786 O cells were higher at lower MOI. The infectiouseffects of Ad5/F11p-GFP in MCF-7 and 786 O, 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 786 O 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 786 O cells than Ad5-GFP did. Conclusion Fiber-modification to Ad5 vector which is derived from chimeric adenovector containing Ad11 p fiber might posses high infection efficiency in tumor cell lines.
出处
《中国医药生物技术》
2015年第2期102-108,共7页
Chinese Medicinal Biotechnology
基金
国家自然科学基金(81372924)
国家高技术研究发展计划(863计划)青年科学家课题(2014AA020515)
