Chimeric 5/35 adenovirus-mediated Dickkopf-1 overexpression suppressed tumorigenicity of CD44+ gastric cancer cells via attenuating Wnt signaling
Chimeric 5/35 adenovirus-mediated Dickkopf-1 overexpression suppressed tumorigenicity of CD44+ gastric cancer cells via attenuating Wnt signaling
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DOI：
10.1007/s00535-012-0711-z
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发表时间：
2013-07-01
影响因子：
6.3
通讯作者：
Wang, Jun
中科院分区：
文献类型：
--
作者：
Wang, Bin;Liu, Jia;Wang, Jun
胃癌干细胞（gastric cancer stem cells，CSCs）需要Wnt信号的激活来维持其自我更新和致瘤性，被认为是有效治疗胃癌的重要靶点。通过血清型5（Ad5）腺病毒或嵌合5/35（Ad5/35）腺病毒递送的基因疗法已显示出治疗各种癌症的前景。本研究采用荧光激活细胞分选法从胃癌原代细胞和胃癌细胞系中分离CD44（+）细胞，建立胃癌CSCs（CD44（+）细胞）的基因治疗方法。在CD44（+）和CD44（-）细胞中检测腺病毒受体的表达。使用Ad5/35（Ad5/35-DKK 1）将有效的Wnt拮抗剂Dickkopf-1（DKK 1）递送到CD44（+）细胞中。CD44（+）细胞中Ad5/35的受体CD46的丰度稍高，而Ad5/35的受体CD46的表达明显降低。因此，CD44（+）细胞对Ad5/35感染敏感，但对Ad5不敏感。Ad5/35-DKK 1将DKK 1引入CD44（+）细胞并有效地灭活内源性Wnt/β-catenin信号传导。DKK 1的过表达抑制了CD44（+）细胞的存活、非贴壁依赖性集落形成和侵袭，而GSK-3特异性抑制剂BIO-acetoxime可以恢复这些功能。更重要的是，DKK 1的引入废除了体内CD44（+）细胞的致瘤性。本研究首次建立了一种基于Ad5/35-DKK 1的抑制肿瘤干细胞Wnt信号通路的新方法，并在临床前实验中证实了胃肿瘤干细胞靶向基因治疗的有效性。
Gastric cancer stem cells (CSCs), which require activation of Wnt signaling to maintain their self-renewal and tumorigenicity, are proposed to be critical targets for effective therapy of gastric carcinomas. Gene therapies that are delivered by adenovirus of serotype 5 (Ad5) or chimeric 5/35(Ad5/35) adenovirus have shown promise for treating various cancers. Here we aimed to develop a gene therapy strategy that targeted gastric CSCs (CD44(+) cells).CD44(+) cells were isolated by fluorescence activated cell sorting from both primary gastric cancer cells and cell lines. Expression of adenovirus receptors was examined in CD44(+) and CD44(-) cells. A potent Wnt antagonist Dickkopf-1 (DKK1) was delivered into CD44(+) cells using Ad5/35 (Ad5/35-DKK1). The therapeutic outcomes were evaluated.Expression of Coxsakievirus adenovirus receptor for Ad5 was significantly reduced, while abundance of CD46, the receptor for Ad5/35, was slightly higher in CD44(+) cells. Accordingly, CD44(+) cells were sensitive to Ad5/35 infection, but not to Ad5. Ad5/35-DKK1 introduced DKK1 into CD44(+) cells and deactivated endogenous Wnt/beta-catenin signaling efficiently. Overexpression of DKK1 inhibited survival, anchorage-independent colony formation, and invasion of CD44(+) cells, which were restored by a GSK-3 specific inhibitor BIO-acetoxime. More importantly, introduction of DKK1 abrogated the tumorigenicity of CD44(+) cells in vivo. However, Ad5/35-DKK1 only showed minimal cytotoxicity to normal tissue-derived cells, L-02 and GES-1.We developed, for the first time, a novel Ad5/35-DKK1-based approach to abrogate Wnt signaling in CSCs and demonstrated that gastric CSC-targeting gene therapy was effective in preclinical experiments.