pVVP3IL-18HN核酸疫苗的构建及其对喉癌细胞杀伤作用的研究

目的通过构建共表达凋亡素基因、新城疫病毒HN基因和白介素18（interleukin-18，IL-18）基因的核酸疫苗，观察联合应用此三种基因对喉癌细胞的杀伤效应。方法在真核表达质粒pVAX1中的CMV启动子下游插入凋亡素基因和含有IL-18HN嵌合基因并携带IRES启动子的基因片段，构建能同时表达三种基因的真核表达质粒pVVP3IL-18HN，经RT—PCR、间接免疫荧光和Western—blot法检测外源基因的表达。采用脂质体介导法将构建的重组质粒pVVP3IL-18HN体外转染Hep-2细胞，运用MTT法检测不同质粒浓度、不同作用时间，该重组质粒对喉癌细胞Hep-2的杀伤率。结果pVVP3IL-18HN可有效杀伤喉癌细胞，且在作用72h、质粒浓度为20μg／ml时杀伤率最高，达61．9％。结论重组质粒pVVP3IL-18HN能有效杀伤Hep-2细胞，可用于喉癌的治疗和研究。

Construction of the nucleic vaccine pVVP3IL-18HN and its antitumor effect on human laryngeal carcinoma

OBJECTIVE: Nucleic vaccine of pVVP3IL-18HN expressing apoptin gene, Newcastle disease virus HN gene and IL-18 gene were constructed to observe the combinative antitumor effect of the above three genes. METHODS: Eukaryotic expression plasmid pVVP3IL-18HN was constructed by inserting apoptin gene and fragment comprising fused IL-18HN gene and IRES promoter into the downstream of CMV promoter of vector pVAX1. The expression of inserted gene was identified by RT-PCR, indirect immunofluorescence and Western-blot. The recombinant plasmid was introduced into Hep-2 cells by liposome, then suppression rate of Hep-2 of different time and different quantity was calculated according to MTT results. RESULT: The recombinant plasmid of pVVP3IL-18HN suppressed Hep-2 successfully and its suppression rate was up to 61.9% with 20 microg/ml, incubation of 72 hours. CONCLUSION: The nucleic vaccine construced pVVP3IL-18HN had antitumor effect on Hep-2. It may can be used to the therapy and research of laryngeal carcinoma.