Drazan K E, Shen X D, Csete M E, Zhang W W, Roth J A, Busuttil R W, Shaked A
Department of Surgery, University of California at Los Angeles.
Surgery. 1994 Aug;116(2):197-203; discussion 203-4.
The aim of this study was to establish a clinically relevant model for gene transfer to liver with an adenoviral vector encoding wild-type p53 as a first step toward use of this class of gene products in the treatment of primary and metastatic liver tumors.
Full-size or 50% hepatectomized rat livers were subjected to asanguineous portal perfusion with a replication-defective adenoviral vector encoding wild-type p53 (Ad5p53), whereas control animals received adenoviral vector encoding Escherichia coli beta-galactosidase (beta-gal) (Ad5LacZ) or Ringer's lactate only. Liver biopsy specimens, blood samples, and liver weight were serially obtained. Gene transfer and expression were confirmed by X-Gal staining for gamma-gal, DNA/RNA polymerase chain reaction, (PCR) and Western blots for p53 and beta-gal. Liver integrity was assessed by histologic findings, serum transaminase levels, and synthetic function.
The gene transfer rate in whole liver and after hepatectomy ranged from 20% to 40%. DNA PCR showed Ad sequences in livers transduced with Ad5p53 and Ad5LacZ. RNA PCR and Western blot confirmed expression and production of recombinant wild-type p53. Liver regeneration was not affected by p53 gene transduction. Liver histologic findings and synthetic function were not different between transduced and control groups.
Ad5p53 gene transfer to full-size or hepatectomized livers is efficient. Liver regeneration and hepatocyte function are unaffected by overexpression of p53. Adenovirus-mediated tumor-suppressor transduction of the liver is a safe and promising adjuvant in cancer gene therapy.
本研究的目的是建立一种与临床相关的模型，用于将编码野生型p53的腺病毒载体导入肝脏，作为在原发性和转移性肝癌治疗中使用这类基因产物的第一步。
将全肝或行50%肝切除的大鼠肝脏用编码野生型p53的复制缺陷型腺病毒载体（Ad5p53）进行无血门静脉灌注，而对照组动物仅接受编码大肠杆菌β-半乳糖苷酶（β-gal）的腺病毒载体（Ad5LacZ）或乳酸林格液。连续获取肝脏活检标本、血样和肝脏重量。通过对γ-半乳糖进行X-Gal染色、对p53和β-半乳糖进行DNA/RNA聚合酶链反应（PCR）以及蛋白质印迹法来确认基因转移和表达。通过组织学检查结果、血清转氨酶水平和合成功能来评估肝脏完整性。
全肝及肝切除后的基因转移率在20%至40%之间。DNA PCR显示在用Ad5p53和Ad5LacZ转导的肝脏中有Ad序列。RNA PCR和蛋白质印迹法证实了重组野生型p53的表达和产生。p53基因转导未影响肝脏再生。转导组和对照组之间的肝脏组织学检查结果和合成功能无差异。
Ad5p53基因向全肝或肝切除后的肝脏转移是有效的。p53的过表达不影响肝脏再生和肝细胞功能。腺病毒介导的肝脏肿瘤抑制因子转导在癌症基因治疗中是一种安全且有前景的辅助治疗方法。
