Cell cycle control pathways act as conditioning factors for TK/GCV sensitivity in pancreatic cancer cells
Cell cycle control pathways act as conditioning factors for TK/GCV sensitivity in pancreatic cancer cells
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DOI：
10.1016/j.bbamcr.2010.06.009
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发表时间：
2010-10-01
影响因子：
5.1
通讯作者：
Fillat, Cristina
中科院分区：
文献类型：
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作者：
Abate-Daga, Daniel;Garcia-Rodriguez, Laura;Fillat, Cristina
自杀系统TK/GCV是一种酶/前药疗法，其涉及将单纯疱疹病毒胸苷激酶基因（TK）的cDNA转移到肿瘤细胞中，然后使细胞对无毒抗病毒药物更昔洛韦敏感。自杀系统TK/GCV虽然被广泛描述，但其作用机制的细节却被掩盖了。为了阐明这个问题，我们进行了旨在确定敏感细胞的关键特征的实验，与对TK/GCV敏感性降低的细胞相比。显示不同程度的敏感性的细胞系在用TK/GCV处理后经历凋亡性细胞死亡。然而，S期延迟几乎仅限于敏感细胞，并且在治疗诱导的抗性模型中受损。在该模型中，鉴定了与诱导抗性相关的差异表达基因。值得注意的是，两个细胞周期相关的基因（CCNE 1和GADD 45）的功能验证为TK/GCV的细胞敏感性的条件。通过显示Chk 1活化与更大TK/GCV细胞毒性相关的实验进一步证明了细胞周期控制的相关性。与Chk 1抑制剂UCN-01的组合处理在敏感细胞中诱导对TK/GCV细胞毒性的拮抗作用，突出了Chk 1活性对TK/GCV作用机制的相关性。这些结果揭示了细胞周期控制途径在TK/GCV系统诱导的细胞毒性中的相关性，将候选基因鉴定为TK/GCV敏感性的调节因子。首次指出Chk 1活化是介导TK/GCV细胞毒作用的关键因素。(C)2010爱思唯尔有限公司版权所有。
The suicide system TK/GCV is an enzyme/prodrug therapy that involves the transfer of the cDNA for the herpes simplex virus thymidine kinase gene (TK) into tumor cells which then sensitizes the cells to the nontoxic antiviral drug ganciclovir. Although extensively characterized, the suicide system TK/GCV conceals the details of its mechanism of action. In order to shed some light on this issue, we conducted experiments designed to identify key features of sensitive cells, as compared to cells that displayed reduced sensitivity to TK/GCV. Cell lines displaying different degrees of sensitivity underwent apoptotic cell death upon treatment with TK/GCV. S-phase delay, however, was almost exclusively restricted to sensitive cells and was impaired in a model of treatment-induced resistance. In this model genes with differential expression associated to induced resistance were identified. Noteworthy, two cell cycle-related genes (CCNE1 and GADD45) were functionally validated as conditioners of cellular sensitivity to TK/GCV. The relevance of cell cycle control was further demonstrated by experiments showing the association of Chk1 activation with greater TK/GCV cytotoxicity. Combination treatment with Chk1 inhibitor UCN-01 induced, in sensitive cells, an antagonistic effect on TK/GCV cytotoxicity highlighting the relevance of Chk1's activity on TK/GCV mechanism of action. These results reveal the relevance of cell cycle control pathways in the cytotoxicity induced by the TK/GCV system identifying candidate genes as conditioners of TK/GCV sensitivity. Moreover it points out, for the first time at Chk1 activation as a key factor to mediate TK/GCV cytotoxicity. (C) 2010 Elsevier B.V. All rights reserved.