新城疫病毒HN基因对肝癌细胞SMMC7721的细胞毒性研究

Cytotoxicity of Newcastle Disease Virus HN Gene on Human Hepatoma Cells SMMC7721

目的:探讨新城疫病毒HN基因对肝癌细胞SMMC7721的杀伤作用及其机制。方法:以脂质体介导方法在体外转染含新城疫病毒HN基因的质粒pVHN于细胞SMMC7721 24 h后,采用MTT法检测细胞活性;3,5-二羟基甲苯测定其唾液酸含量的变化;丫啶橙/溴化乙锭(AO/EB)染色,荧光显微镜观察细胞形态学改变;以及FCM检测病毒HN基因和HLA-A,B, C的表达。结果:体外转染pVHN能显著地降低细胞表面唾液酸含量(P<0.05),有效地杀伤肿瘤细胞SMMC7721;荧光显微镜观察可见典型的细胞死亡形态学改变;实验组细胞与对照组比较HN表达差异显著,HLA-A,B,C表达上调。结论:HN基因在体外能够显著降低肿瘤细胞表面唾液酸含量,同时,使其高表达HN抗原,上调HLA-A,B,C表达,从而,可能增强了肿瘤细胞的抗原性和免疫识别,诱导了细胞SMMC7721死亡。

Objective： To investigate the mechanism on cytotoxicity of human hepatoma cells SMMC7721 induced by NDV HN gene. Methods： 24 h after transfected with the mixture of NDV HN gene and liposome in vitro, SMMC7721 cell mortality rate was measured through MTT staining ; the content of cell surface sialic acid was measured; the death pattern of SMMC7721 cells was detected by staining with AO/EB ; expressions of HN gene and HLA-A, B, C was measured by FCM. Results： Transfected into SMMC7721 cells 24 h later, pVHN reduced the surface sialic acid of SMMC7721 cells（P 〈 0.05 ） and killed the cells significantly in vitro ; the difference of HN gene expression was significantly between experimental groups and control groups, and the HLA-A, B, C expression was upregulated. Conclusions： The HN gene plays an important role in the reduction of sialic acid of SMMC7721 cells and caused the cell death. The expressions of HN gene was significant and HLA-A, B, C expression was upregnlated after HN gene tansfeeted into SMMC7721 in vitro. Therefore, immunogenicity and immunoregonization of tumor cells might be increased, so the SMMC7721 cells was killed.