辣根过氧化物酶/吲哚三醋酸酶前药对鼻咽癌细胞的杀伤作用  

Indole-3-Acetic acids and horseradish peroxidase:the novel Prodrug/Enzyme combination for human nasopharyngeal carcinoma

作　　者：苗北平[1] 张蕊石[2] 孟庆国[1] 王晓彬[1] 孙焕吉[1] 林峰[1] 卢永田[1] 

机构地区：[1]深圳市第二人民医院深圳大学第一附属医院耳鼻咽喉科,广东深圳518035 [2]深圳市第二人民医院深圳大学第一附属医院眼科,广东深圳518035

出　　处：《山东大学耳鼻喉眼学报》2009年第3期9-14,共6页Journal of Otolaryngology and Ophthalmology of Shandong University

基　　金：深圳市科技局重点资助课题(200901008)

摘　　要：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 ability of the recombinant replication-defective adenovirus vector adv5. oriP. HRP to target the Horseradish Peroxidase （HRP） gene into EBV-positive nasopharyngeal carcinoma cells to confer sensitivity to Indole-3-Acetic Acids （IAA） to the cells, and to investigate the killing ability of HRP/IAA to nasopharyngeal carcinoma cells under oxic and anoxic conditions and the bystander effect. Methods C666-1, KS1 and CNE-2Z were incubated with adv5.oriP.HRP, then IAA was added. MTT assay was used to evaluate the cytotoxicity of the suicide gene therapy whether in oxic or anoxic conditions. Results When being infected with a recombinant adenovirns carrying the HRP gene, high expression of HRP can be seen in C666- 1 cells, which were killed by IAA. However, the infected KS1 and CNE-2Z cells remained resistant to IAA and no HRP protein were found. HRP/IAA can kill EBV-positive C666-1 cells in oxic and anoxic conditions. In addition, a bystander killing effect was demonstrated against C666-1 cells when only 20% of the adv5. oriP. HRP-infected cells were mixed with the uninfected cells, and the eradication of cells can almost be achieved. Conclusion These results demonstrate that the oriP sequence can achieve high levels of gene expression specifically targeted to EBV-positive NPC cells in the context of the adv vector. HRP/IAA is a novel Prodrug/Enzyme Combination for NPC. This has now provided the tumor-specific expression system from which additional interventions can be evaluated in future treatment strategies for patients with nasopharyngeal cancer.

关 键 词：辣根过氧化物酶/吲哚三醋酸酶 酶前药 乏氧/有氧状态 旁观者效应 

分 类 号：R739.65[医药卫生—肿瘤] Q78[医药卫生—临床医学]