State Key Laboratory of Virology, School of Basic Medicine, Wuhan University, Wuhan, People's Republic of China.
Cancer Gene Ther. 2012 Feb;19(2):77-83. doi: 10.1038/cgt.2011.65. Epub 2011 Sep 30.
Clinical efficacy of current therapies for hepatocellular carcinoma (HCC) treatment is limited. Indole-3-acetic acid (IAA) is non-toxic for mammalian cells. Oxidative decarboxylation of IAA by horseradish peroxidase (HRP) leads to toxic effects of IAA. The purpose of this study was to investigate the effects of a novel gene-targeted enzyme prodrug therapy with IAA on hepatoma growth in vitro and in vivo mouse hepatoma models. We generated a plasmid using adenovirus to express HRP isoenzyme C (HRPC) with the HCC marker, alpha-fetoprotein (AFP), as the promoter (pAdv-AFP-HRPC). Hepatocellular cells were infected with pAdv-AFP-HRPC and treated with IAA. Cell death was detected using MTT assay. Hepatoma xenografts were developed in mice by injection of mouse hepatoma cells. The size and weight of tumors and organs were evaluated. Cell death in tumors was assessed using hematoxylin and eosin-stained tissue sections. HRPC expression in tissues was detected using Reverse Transcriptase-Polymerase Chain Reaction. IAA stimulated death of hepatocellular cells infected with pAdv-AFP-HRPC, in a dose- and time-dependent manner, but not in control cells. Growth of hepatoma xenografts, including the size and weight, was inhibited in mice treated with pAdv-AFP-HRPC and IAA, compared with that in control group. pAdv-AFP-HRPC/IAA treatment induced cell death in hepatoma xenografts in mice. HRPC gene expressed only in hepatoma, but not in other normal organs of mice. pAdv-AFP-HRPC/IAA treatment did not cause any side effects on normal organs. These findings suggest that pAdv-AFP-HRPC/IAA enzyme/prodrug system may serve as a strategy for HCC therapy.
目前治疗肝细胞癌（HCC）的疗法疗效有限。吲哚-3-乙酸（IAA）对哺乳动物细胞无毒。辣根过氧化物酶（HRP）氧化脱羧 IAA 会导致 IAA 的毒性作用。本研究旨在探讨用 IAA 进行新型基因靶向酶前药治疗对肝癌细胞在体和体内小鼠肝癌模型生长的影响。我们使用腺病毒生成了一个质粒，该质粒用 HCC 标志物甲胎蛋白（AFP）作为启动子表达 HRP 同工酶 C（HRPC）（pAdv-AFP-HRPC）。用 pAdv-AFP-HRPC 感染肝癌细胞并给予 IAA 处理。用 MTT 法检测细胞死亡。通过注射小鼠肝癌细胞在小鼠中建立肝癌异种移植模型。评估肿瘤和器官的大小和重量。通过苏木精和伊红染色组织切片评估肿瘤中的细胞死亡。用逆转录-聚合酶链反应检测组织中的 HRPC 表达。IAA 以剂量和时间依赖的方式刺激感染了 pAdv-AFP-HRPC 的肝癌细胞死亡，但对对照细胞没有作用。与对照组相比，用 pAdv-AFP-HRPC 和 IAA 治疗的小鼠肝癌异种移植的生长，包括大小和重量均受到抑制。pAdv-AFP-HRPC/IAA 治疗在小鼠肝癌异种移植中诱导了细胞死亡。HRPC 基因仅在肝癌中表达，而不在其他正常小鼠器官中表达。pAdv-AFP-HRPC/IAA 治疗对正常器官没有任何副作用。这些发现表明 pAdv-AFP-HRPC/IAA 酶/前药系统可能成为 HCC 治疗的一种策略。
