目的：经Beagle犬肝脏血管注射重组腺病毒介导反义c-myc基因 (Ad-ASmyc )载体，观测其体内分布及毒副作用，以评价Ad-ASmyc治疗肝癌的临床前期安全性。方法:选择4只体重8～10 kg 健康Beagle犬，全麻后开腹，经肝动脉或门静脉注射Ad-ASmyc，注射Ad-ASmyc前及注射后第3,7,14,21天取肝组织及抽取静脉血化验，PCR检测Ad-ASmyc在各器官组织中的分布，常规切片观察各器官病理变化，ELISA方法检测血中抗腺病毒抗体的产生。结果: 经Beagle犬肝脏血管注射后，Ad-ASmyc可持续转导正常肝细胞达3周，实验犬一般情况好，血、肝、肾功能无明显异常，在肝脏、脾脏、肾脏、胃、心脏、皮肤中可检测到重组腺病毒的分布，镜下可见Ad-ASmyc剂量依赖性的肝组织轻微的炎症反应，注射后7 d血中有抗腺病毒载体的抗体产生，14 d达高峰，21 d开始下降。结论:经Beagle犬肝动脉或门静脉途径注射Ad-ASmyc均可转导至肝细胞，Ad-ASmyc对实验犬的毒副作用较轻

Objective: To assess the preclinical safety of recombinant adenovirus-mediated antisense c-myc infustion of the hepatic vascular of Beagle dogs. Methods: Four 8 to 10 kg healthy Beagle dogs underwent hepatic artery or portal vein cannulation infusion of 1×10 10～1×10 11 plaque forming units recombinant adenovirus vectors. Serial sequenced liver biopsies were taken for microscopic examination and PCR analysis. Venous blood samples were obtained from the dogs for liver, renal function tests and hematology analysis after infusion of days 0, 3, 7, 14, and 21. PCR was used to screen the vital organs for the presence of adenovirus DNA. Microscopic examination of the vital organs was performed to observe the pathogenicity of Ad-ASmyc. ELISA was performed to assay the neutralizing antibody to adenovirus vectors. Results：Ad-ASmyc could infect both kinds of normal human fetal lung cell line 2BS and normal human hepatocyte line L02 effectively, but could not inhibit their growth in vitro. Results of liver, renal function and hematology values were within normal ranges. The adenovirus vectors were present in the liver, spleen, heart, kidney, stomach and skin at days 14 or 21. Microscopic examination revealed no cytopathic effects in distant organs, as was gross pathology, despite the presence of vector. Ad-ASmyc was transferred into hepatocytes persistently and induced dose-dependent inflammation response. Production of anti-adenovirus antibodies appeared at days 7, reached high-level at days 14 and declined at days 21.Conclusions：Ad-ASmyc can be transferred into hepatocellulars via hepatic artery or portal vein and generates no obvious toxicity in Beagle dogs, which suggests the safety of Ad-ASmyc infused into the hepatic vascular for gene therapy.

R730.5

国家863高科学技术发展基金资助项目（Z20-01-02）