双融膜嗜肿瘤病毒对肺腺癌细胞的杀伤效果研究

Evaluation of the antitumor activity of double fusogenic virus against A549 pulmonary adenocarcinoma

目的:构建双融膜嗜肿瘤病毒,并观察其杀伤肺癌细胞的效果。方法:将长臂猿白血病病毒融膜糖蛋白基因(Fusogenic glycoprotein gene of gibbon ape leukemia virus,GALV.fus)构建入嗜肿瘤Ⅰ型单纯疱疹病毒(Herpes simplex virus 1,HSV-1)中,使其成为具有双重融细胞膜作用的嗜肿瘤病毒(Doubly fusogenic oncolytic herpes simplex virus,Synco-2D)。与单一的重组嗜肿瘤病毒HSV-1比较,在显微镜下观察两种嗜肿瘤病毒融细胞现象,并观察其体外杀伤肺腺癌细胞A549的效率及体内对肿瘤生长的抑制作用。结果:显微镜下发现,双融膜病毒融细胞作用更明显,大量多核巨细胞形成,细胞结构模糊,细胞溶解。体外实验显示,两种病毒都有明显的杀死肿瘤细胞效果,双融膜嗜肿瘤病毒杀伤肿瘤细胞率较单一的HSV-1明显增强。体内动物实验显示双融膜嗜肿瘤病毒对肿瘤组织的生长有显著的抑制作用。结论:将融膜糖蛋白基因GALV.fus构建入嗜肿瘤Ⅰ型单纯疱疹病毒中,可以显著增强病毒杀伤肺腺癌细胞效力。

Objective：To construct double fusogenic oncolytic virus and evaluate its antitumor activity against A549. Methods： The double fusogenic oncolytic virus was incorporated with both fusion mechanisms into a single oncolytic HSV throug insertion of the hyperfusogenic glycoprotein gene of gibbon ape leukemia virus（ GALV.fus ） into HSV-1. Syncytial phenotype was observed with microscope. Double fusogenic virus antitumor activity against A549, compared with simple oncolytic HSV, was evaluated in vitro and vivo. Results：The A549 cell infected with the two kind of oncolytic virus showed syncytial phenotype respectively. A distinctive syncytial phenotype was observed with the double fusogenic oncolytic HSV（Synco-2D） under microscope. In vitro characterization of Synco-2D showed that this virus produced a distinctive antitumor ability, compared with that of a nonfusogenic virus. When oncolytic virus was injected directly into the abdominal cavity of mice bearing A549 ceils, Synco-2D eradicated tumor masses significantly as compared with single oncolytic HSV. Conclusion： A double fusogenic oncolytic HSV can significantly increase killing ability against lung adenoca rcinoma cell A549.