Vergara Daniele, de Domenico Stefania, Maffia Michele, Piro Gabriella, Di Sansebastiano Gian-Pietro
Department of Biological and Environmental Sciences and Technologies (DiSTeBA), University of Salento, Campus ECOTEKNE, S.P. 6, Lecce-Monteroni, Lecce 73100, Italy.
Institute of Science of Food Production, National Research Council (CNR), Lecce 73100, Italy.
Int J Mol Sci. 2015 Jan 20;16(1):2174-86. doi: 10.3390/ijms16012174.
In this work we explored the possibility of using genetically modified Arabidopsis thaliana plants as a rapid and low-cost screening tool for evaluating human anticancer drugs action and efficacy. Here, four different inhibitors with a validated anticancer effect in humans and distinct mechanism of action were screened in the plant model for their ability to interfere with the cytoskeletal and endomembrane networks. We used plants expressing a green fluorescent protein (GFP) tagged microtubule-protein (TUA6-GFP), and three soluble GFPs differently sorted to reside in the endoplasmic reticulum (GFPKDEL) or to accumulate in the vacuole through a COPII dependent (AleuGFP) or independent (GFPChi) mechanism. Our results demonstrated that drugs tested alone or in combination differentially influenced the monitored cellular processes including cytoskeletal organization and endomembrane trafficking. In conclusion, we demonstrated that A. thaliana plants are sensitive to the action of human chemotherapeutics and can be used for preliminary screening of drugs efficacy. The cost-effective subcellular imaging in plant cell may contribute to better clarify drugs subcellular targets and their anticancer effects.
在这项工作中，我们探索了利用基因改造的拟南芥植物作为一种快速且低成本的筛选工具，用于评估人类抗癌药物的作用和疗效。在此，我们在植物模型中筛选了四种在人类中具有已验证抗癌效果且作用机制不同的抑制剂，以研究它们干扰细胞骨架和内膜网络的能力。我们使用了表达绿色荧光蛋白（GFP）标记的微管蛋白（TUA6-GFP）的植物，以及三种可溶性GFP，它们分别通过不同的方式分选，一种定位于内质网（GFPKDEL），另外两种分别通过依赖COPII（AleuGFP）或不依赖COPII（GFPChi）的机制在液泡中积累。我们的结果表明，单独或联合测试的药物对包括细胞骨架组织和内膜运输在内的所监测细胞过程产生了不同的影响。总之，我们证明了拟南芥植物对人类化疗药物的作用敏感，可用于药物疗效的初步筛选。植物细胞中具有成本效益的亚细胞成像可能有助于更好地阐明药物的亚细胞靶点及其抗癌效果。
