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目的研究adv5.oriP.HRP对鼻咽癌细胞靶向性转染和HRP/IAA对鼻咽癌细胞的杀伤作用。方法将adv5.oriP.HRP感染C666-1,KS1和CNE-2Z细胞,加入IAA,用MTT法评估细胞杀伤效果。建立乏氧有氧条件,检测HRP/IAA在乏氧状态下鼻咽癌细胞的杀伤能力及旁观者效应。结果adv5.oriP.HRP能靶向性在EBV阳性的鼻咽癌细胞内高效转染和表达。HRP/IAA对鼻咽癌细胞有强大的杀伤作用,在一定范围内杀伤能力随孵育时间和病毒载体及前药的浓度增高而增加。在乏氧状态下,HRP/IAA有同样的杀伤能力。当感染细胞占总细胞的10%左右时,65%的肿瘤细胞死亡,当比例达到20%时,90%以上的细胞死亡,显示HRP/IAA有很强的旁观者效应。结论adv5.oriP.HRP能在EBV病毒阳性的细胞内靶向性表达;HRP/IAA在乏氧和有氧状态下均有强大的杀伤能力和旁观者效应。adv5.oriP.HRP/IAA是理想的病毒介导的酶前药基因组合。
Objective To study the targeted transfection of adv5.oriP.HRP on nasopharyngeal carcinoma cells and the killing effect of HRP / IAA on nasopharyngeal carcinoma cells. Methods C666-1, KS1 and CNE-2Z cells were infected with adv5.oriP.HRP, IAA was added, and the cell killing effect was evaluated by MTT assay. Establishment of hypoxia and aerobic conditions, detection of HRP / IAA nasopharyngeal carcinoma cells in hypoxia state cytotoxicity and bystander effect. Results adv5.oriP.HRP can be targeted in EBV-positive nasopharyngeal carcinoma cells efficiently transfected and expressed. HRP / IAA has a strong killing effect on nasopharyngeal carcinoma cells. In a certain range, the killing ability of HRP / IAA increases with the incubation time and the concentration of viral vectors and prodrugs. Under hypoxic conditions, HRP / IAA has the same killing ability. When infected cells make up about 10% of the total cells, 65% of the tumor cells die. When the proportion reaches 20%, more than 90% of the cells die, indicating a strong bystander effect of HRP / IAA. Conclusions adv5.oriP.HRP can be targeted expressed in EBV-positive cells. HRP / IAA has strong cytotoxicity and bystander effect both in hypoxia and aerobic state. adv5.oriP.HRP / IAA is the ideal viral-mediated enzyme prodrug gene combination.