Hypoxia/hepatoma dual specific suicide gene expression plasmid delivery using bio-reducible polymer for hepatocellular carcinoma therapy.
Hypoxia/hepatoma dual specific suicide gene expression plasmid delivery using bio-reducible polymer for hepatocellular carcinoma therapy.
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DOI：
10.1016/j.jconrel.2013.06.033
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发表时间：
2013-10-10
期刊：
影响因子：
--
通讯作者：
Kim SW
中科院分区：
文献类型：
--
作者：
Kim HA;Nam K;Lee M;Kim SW
基因治疗被认为是肝细胞癌（HCC，也称肝癌）治疗的一种有前途的替代策略。为了实现成功和安全的基因治疗，需要严格调控基因表达以最小化正常组织中的副作用。在本研究中，我们开发了一种新的缺氧和肝癌双特异性基因表达载体。使用生物可还原聚合物PAM-ABP将构建的载体转染到各种细胞系中。首先，构建了具有AFP启动子和增强子的肝癌组织特异性基因表达载体pAFPS-Luc或pAFPL-Luc。然后，通过插入促红细胞生成素（Epo）增强子构建pEpo-AFPL-Luc，用于低氧癌症特异性基因表达。体外转染实验表明，pEpo-AFPL-Luc转染的肝癌细胞在缺氧条件下基因表达增加。为了证实双特异性载体的治疗效果，引入单纯疱疹病毒胸苷激酶基因（HSV-TK）用于杀伤癌细胞。在更昔洛韦（GCV）前药的存在下，将pEpo-AFPL-TK转染到肝癌细胞系中。Caspase-3/7、MTT和TUNEL检测结果表明，转染pEpo-AFPL-TK的肝癌细胞在缺氧条件下的死亡率明显高于对照组。因此，含Epo增强子和AFP启动子的低氧/肝癌双特异性基因表达载体可能用于肝癌特异性基因治疗。
Gene therapy is suggested as a promising alternative strategy of hepatocellular carcinoma (HCC, also called hepatoma) therapy. To achieve a successful and safe gene therapy, tight regulation of gene expression is required to minimize side-effects in normal tissues. In this study, we developed a novel hypoxia and hepatoma dual specific gene expression vector. The constructed vectors were transfected into various cell lines using bio-reducible polymer, PAM-ABP. First, pAFPS-Luc or pAFPL-Luc vector was constructed with the alpha-fectoprotein (AFP) promoter and enhancer for hepatoma tissue specific gene expression. Then, pEpo-AFPL-Luc was constructed by insertion of the erythropoietin (Epo) enhancer for hypoxic cancer specific gene expression. In vitro transfection assay showed that pEpo-AFPL-Luc transfected hepatoma cell increased gene expression under hypoxic condition. To confirm the therapeutic effect of dual specific vector, herpes simplex virus thymidine kinase gene (HSV-TK) was introduced for cancer cell killing. The pEpo-AFPL-TK was transfected into hepatoma cell lines in the presence of ganciclovir (GCV) pro-drug. Caspase-3/7, MTT and TUNEL assays elucidated that pEpo-AFPL-TK transfected cells showed significant increasing of death rate in hypoxic hepatoma cells compared to controls. Therefore, the hypoxia/hepatoma dual specific gene expression vector with the the Epo enhancer and AFP promoter may be useful for hepatoma specific gene therapy.
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