经肝动脉或瘤体注射单纯疱疹病毒-胸苷激酶基因治疗兔肝癌

目的　观察瘤体内直接注射或经肝动脉注射载有单纯疱疹病毒胸苷激酶 (HSV TK)基因的EB病毒表达质粒 pDR2 /TK、丙氧鸟苷 (GCV )对原位兔肝癌的治疗效果。 方法　制作兔原位肝癌模型 (VX2 ) ,瘤体注射或经肝动脉注射质粒 pDR2 /TK ,腹腔注射GCV连续 10d。RT PCR检测肝癌HSV TK表达 ;螺旋CT监测肝癌大小 ,并观察兔存活时间。结果TK基因导入 10d后 ,直接注射组TK在肝癌组织强表达 ,癌旁组织弱表达 ,正常肝组织不表达 ;肝动脉注射组TK基因在肝癌表达稍强于癌旁及正常肝组织。直接注射组肿瘤大小 ( 3 .5 5± 0 .3 9)cm ,与经肝动脉注射+肝动脉结扎组肿瘤大小 ( 3 .70± 0 .3 7)cm无明显差别 (P >0 .0 5 ) ,但均明显小于对照组。瘤体直接注射TK基因 +GCV治疗组动物平均存活时间 ( 5 9.8± 3 .3 )d、肝动脉注射组 +肝动脉结扎组( 5 4.8± 4.5 )d明显长于各对照组 (P均 <0 .0 1)。结论　对实验性兔肝癌 ,瘤体内直接注射或经肝动脉注射导入治疗基因后 ,HSV TK/GCV系统具有较好的治疗效果。

Effects of gene therapy on the experimental rabbit hepatocellular carcinoma by direct intratumoral or intra-hepatic artery injection of HSV-TK gene

Objective To investigate the therapeutic effects of herpes simplex virus thymidine kinase (HSV-TK) gene plus GCV treatment on the experimental rabbit hepatocellular caricinoma by two routes of HSV-TK gene delivery:direct intratumoral (IT) and intra-hepatic artery (IHA) injection.Methods Using VX2 liver carcinoma model,the experimental rabbit orthotopic hepatoma was made.Under laparotomy,Epstein-Barr virus based expression plasmid pDR2/TK carrying HSV-TK gene were delivered into rabbit hepatoma by IT or IHA injection,then intraperitoneal treatment with ganciclovir (GCV) was done for 10 successive days.RT-PCR and spiral CT were performed to examine the expression of HSV-TK gene in rabbit hepatoma and tumor size respectively.The antitumoral efficacy of gene therapy was evaluated with tumor size and rabbit survival time.Results After transfection of TK gene for 10 days by two routes,the expression of TK gene in rabbit hepatoma was all relatively strong,in the group of IT that in nomal liver tissue and in tissue near the tumor was weak even negative,but in the group of IHA that was moderate.In experimental group with TK gene transfection (IT and IHA) plus GCV treatment,the tumor size was significantly less than that in any other control group,and the rabbit survival time was also significantly prolonged nearly two times (P<0.01).Conclusion Either IT or IHA injection can lead to effective gene delivery and gene expression,and the HSV-TK/GCV system was obviously effective to treat experimental rabbit hepatocellular caricinoma.