新城疫病毒pIRHN核酸疫苗构建和表达及对肿瘤细胞的影响

目的：研究NDV HN基因抗肿瘤作用及其可能机制。方法：以 pIRES1neo为表达载体构建了 NDV HN基因的pIRHN核酸疫苗，在体外转染HeLa细胞，用间接免疫荧光和Western blot检测pIRHN在真核细胞中表达状况，用荧光显微镜、DNA琼脂糖电泳及TUNEL染色等方法，检测HN基因导致细胞死亡的类型；用3，5-二羟基甲苯法测定HeLa细胞唾液酸含量的变化。结果：pIRHN 转染HeLa细胞后，能够在真核细胞中表达，能促进肿瘤细胞死亡，其死亡方式主要以诱导细胞凋亡为主，pIRHN使 HeLa细胞唾液酸含量减少。结论：用 NDV HN基因所构建的核酸疫苗能够在真核细胞中高效表达，表达的 HN蛋白主要位于胞膜，胞浆中亦有 HN蛋白表达；pIRHN具有抗肿瘤作用，可能通过其表达产物与肿瘤细胞唾液酸受体的相互作用，发挥其抗肿瘤作用。本实验为迸一步阐明NDV抗肿瘤作用机制提供了理论依据。

Construction and expression of NDV pIRHN nucleic acid vaccine and its effect on the tumor cells

Abstract Objective:To explore antitumor effect of NDV HN. Methods: pIRHN nucleic acid vaccin was constructed and was transfectedto Hela cells. Westem blot was used to analyssize the expression of pIRHN nucleic acid vaccine in eukarytic cell. The mode of cell death was de-tected by fluorescence microscope, gel electrophoresis and TUNEL assay. The effect of pIRHN nucleic acid vaccine on the contents of siaha acidin the Hela cell was examined. Results: pIRHN nucleic acid vaccine could be expressed in the eukarytic cell. pIRHN could induced apoptosisafter HeLa cells were transfected. The effect of antitumor responses of pIRHN was correlated with the contents of sialic acid in tumor cells. Con-clusion: The reults of our experiment will provide the theory evidence for the anti tumor mechamism of NDV.