Functional Gene Knockout of NRF2 Increases Chemosensitivity of Human Lung Cancer A549 Cells In Vitro and in a Xenograft Mouse Model.
Functional Gene Knockout of NRF2 Increases Chemosensitivity of Human Lung Cancer A549 Cells In Vitro and in a Xenograft Mouse Model.
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DOI：
10.1016/j.omto.2018.10.002
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发表时间：
2018-12-21
期刊：
影响因子：
--
通讯作者：
Kmiec EB
中科院分区：
文献类型：
--
作者：
Bialk P;Wang Y;Banas K;Kmiec EB
最近的研究指出，肺癌耐药性的演变至少部分集中在参与控制外排或药物失活的各种基因的上调上。这些基因中最重要的是核因子红细胞2相关因子（NRF 2），其被认为是参与对氧化和/或亲电应激的细胞应答的100-200个靶基因的主调节因子。随着人们越来越关注癌症治疗组合方法的开发，我们利用CRISPR/Cas9通过破坏NRF 2核输出信号（内斯）结构域来禁用肺癌细胞中的NRF 2基因;从表型上看，该蛋白质在很大程度上被阻止翻译后进入细胞核。在组织培养中，发现具有该基因敲除的细胞具有降低的增殖表型，并且对化疗剂如顺铂和卡铂更敏感。这些观察结果在异种移植小鼠模型中得到证实，其中纯合敲除细胞的增殖速率低于野生型细胞，即使在不存在药物治疗的情况下也是如此。肿瘤生长被阻止了16天，在接受CRISPR指导的基因编辑和化疗的联合作用的样品中观察到肿瘤体积急剧减少。
Recent studies point to the evolution of drug resistance in lung cancer as being centered, at least in part, on the upregulation of various genes involved in controlling efflux or drug inactivation. Among the most important of these genes is Nuclear Factor Erythroid 2-Related Factor (NRF2), considered the master regulator of 100–200 target genes involved in cellular responses to oxidative and/or electrophilic stress. With increased focus on the development of combinatorial approaches for cancer treatment, we utilized CRISPR/Cas9 to disable the NRF2 gene in lung cancer cells by disrupting the NRF2 nuclear export signal (NES) domain; phenotypically, the protein is largely blocked from transiting into the nucleus after translation. In tissue culture, cells with this gene knockout were found to have a reduced proliferation phenotype and are more sensitive to chemotherapeutic agents, such as cisplatin and carboplatin. These observations were confirmed in xenograft mouse models wherein the homozygous knockout cells proliferate at a slower rate than the wild-type cells, even in the absence of drug treatment. Tumor growth was arrested for a period of 16 days, with a dramatic decrease in tumor volume being observed in samples receiving the combined action of CRISPR-directed gene editing and chemotherapy.
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影响因子：
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作者：
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通讯作者：
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