Expression of HIF-1α ODD domain fused canine caspase 3 by EGFR promoter-driven adenovirus vector induces cytotoxicity in canine breast tumor cells under hypoxia
Expression of HIF-1α ODD domain fused canine caspase 3 by EGFR promoter-driven adenovirus vector induces cytotoxicity in canine breast tumor cells under hypoxia
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DOI：
10.1007/s11259-016-9664-7
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发表时间：
2016-12-01
影响因子：
2.2
通讯作者：
Ikeda, Teruo
中科院分区：
文献类型：
--
作者：
Okamoto, Mariko;Asamura, Ai;Ikeda, Teruo
腺病毒载体广泛应用于肿瘤基因治疗。然而，与人类患者相比，在用Ad载体转导的犬肿瘤细胞中，关于基因转导效率或细胞特异性细胞毒性的信息相对有限。由于表皮生长因子受体（EGFR）是高度表达的犬乳腺肿瘤细胞，我们试图开发一种广告载体的基础上的RGD纤维突变型腺病毒载体（AdRGD），表达犬caspase 3的EGFR启动子的控制下。本研究的目的是实现高转导效率的转基因表达限制在犬乳腺肿瘤细胞。使用EGFR启动子驱动的AdRGD，我们能够将转基因表达限制在犬乳腺肿瘤细胞中，而在正常细胞中没有表达的证据。EGFR启动子驱动的携带犬caspase 3基因的AdRGD转导的犬乳腺癌细胞具有细胞毒活性。我们构建了第二个表达EGFR启动子控制下的氧依赖性降解（ODD）-caspase 3的AdRGD载体;融合蛋白含有与caspase 3融合的缺氧诱导因子-1 α（HIF-1 α）的ODD结构域的核心部分。与常氧条件相比，用表达ODD-caspase 3的EGFR启动子驱动的AdRGD转导犬乳腺肿瘤细胞，在缺氧条件下诱导了更高的细胞死亡率。结果表明，EGFR启动子驱动的AdRGD载体将在肿瘤特异性转基因表达和安全的癌症基因治疗狗的价值。
Adenovirus (Ad) vectors are widely used in cancer gene therapies. However, compared to human patients, relatively limited information is available on gene transduction efficiency or cell-specific cytotoxicity in canine tumor cells transduced with Ad vectors. Since epidermal growth factor receptor (EGFR) is highly expressed on canine breast tumor cells, we sought to develop an Ad vector based on the RGD fiber-mutant adenovirus vector (AdRGD) that expresses canine caspase 3 under the control of EGFR promoter. The aims of this study were to achieve high transduction efficiency with transgene expression restricted to canine breast tumor cells. Using EGFR promoter-driven AdRGD, we were able to restrict transgene expression to canine breast tumor cells with no evidence of expression in normal cells. Canine breast tumor cells transduced with EGFR promoter-driven AdRGD carrying canine caspase 3 gene showed cytotoxic activity. We constructed a second AdRGD vector that expressed oxygen-dependent degradation (ODD)-caspase 3 under the control of the EGFR promoter; the fusion protein contains a core part of the ODD domain of hypoxia inducible factor-1 alpha (HIF-1 alpha) fused to caspase 3. Transduction of canine breast tumor cells with EGFR promoter-driven AdRGD expressing ODD-caspase 3 induced a higher rate of cell death under hypoxic conditions compared with under normoxia. The results indicate that the EGFR promoter-driven AdRGD vectors will be of value for tumor-specific transgene expression and safe cancer gene therapy in dogs.