GALV致融性糖蛋白转导肺腺癌细胞的抗肿瘤实验研究

Antitumor effect of GALV membrane fusion glycoprotein on lung adenocarcinoma in vivo and in vitro

目的探讨特异性启动子控制下嗜肿瘤Ⅰ型单纯疱疹病毒(HSV-Ⅰ)载体介导的长臂猿白血病病毒致融性外膜糖蛋白(GALV.fus)基因转导肺腺癌细胞的抗肿瘤作用。方法将重组HSV-Ⅰ载体质粒通过脂质体导入非洲绿猴肾细胞(Vero)中包装制备重组HSV-Ⅰ病毒,用来转染肺腺癌细胞(A549)和正常人胚胎成纤维细胞(HFL-ⅠGNHu5)及裸鼠皮下肺癌转移模型,观察其体外及体内抗肿瘤作用及细胞毒性作用。统计学方法采用t检验。结果成功包装重组HSV-Ⅰ病毒,BALB/c裸鼠皮下接种A549细胞2周后均有肿瘤生长,成瘤率为100%。重组单纯疱疹病毒能特异性转染A549细胞,治疗组及对照组细胞存活率分别为20%及70%,病毒对A549细胞有明显的杀灭作用且对裸鼠皮下肺癌转移灶抑制明显。结论GALV.fus基因对肺癌细胞有强效的抗肿瘤作用,为临床肺癌治疗探索出一种新型基因治疗方法。

Objective To investigate the antitumor effect of special promoter-controlled Gibbon ape leukemia virus membrane fusion glycoprotein （ GALV. fus） mediated by type Ⅰ herpes simplex virus （ HSV-Ⅰ ） on lung adenocarcinoma. Methods Recombinant HSV-Ⅰ plasmids encoding GALV. fus was introduced into green monkey kidney cells（Vero） by liposome to amplify the virus, and then the virus was transfected into lung adenocarcinoma （A549） , human fetal fibroblasts （HFL-Ⅰ GNHu 5 ） and human lung adenocarcinoma xeno- grafts which were established in nude mice subcutaneously to observe antitumor and cytotoxic effect in vitro and in vivo; Recombined cytomegalovirus （CMV） containing GALV. fus or enhanced green fluorescence protein were served as control. Results Recombinant HSV-Ⅰ virus were packed successfully. Heterotransplantative tumourigenicity of the tumour was 100% in nude mice after A549 cells were inoculated. Recombinant HSV-Ⅰ virus exerted obvious antitumor effects in vitro, with relative survival rate of 23%, while for CMV virus containing GALV. fus, the rate was 20% , and for CMV virus encoding EGFP, the rate was 68%. Recombinant HSV-Ⅰ virus also showed striking antitumor effect on the implanted tumor. Conclusion GALV. fus has powerful effect against lung cancer in vitro and in vivo and maybe a promising candidate for gene therapy.