Macovei Anca, Pagano Andrea, Sabatini Maria Elisa, Grandi Sofia, Balestrazzi Alma
Department of Biology and Biotechnology 'L. Spallanzani', University of Pavia, via Ferrata 9, 27100 Pavia, Italy.
Genes (Basel). 2018 Mar 28;9(4):186. doi: 10.3390/genes9040186.
The hTdp1 (human tyrosyl-DNA phosphodiesterase 1) inhibitor NSC120686 has been used, along with topoisomerase inhibitors, as a pharmacophoric model to restrain the Tdp1 activity as part of a synergistic treatment for cancer. While this compound has an end-point application in medical research, in plants, its application has not been considered so far. The originality of our study consists in the use of hTdp1 inhibitor in cells, which, unlike human cells, contain two genes. Hence, the purpose of this study was to test the hTdp1 inhibitor NSC120686 as an exploratory tool to investigate the plant genes, since their characterization is still in incipient phases. To do so, calli were exposed to increasing (75, 150, 300 μM) concentrations of NSC120686. The levels of cell mortality and DNA damage, measured via diffusion assay and comet assay, respectively, were significantly increased when the highest doses were used, indicative of a cytotoxic and genotoxic threshold. In addition, the NSC120686-treated calli and untreated -depleted calli shared a similar response in terms of programmed cell death (PCD)/necrosis and DNA damage. Interestingly, the expression profiles of and genes were differently affected by the NSC120686 treatment, as was upregulated while was downregulated. The NSC120686 treatment affected not only the genes but also other genes with roles in alternative DNA repair pathways. Since the expression patterns of these genes were different than what was observed in the -depleted plants, it could be hypothesized that the NSC120686 treatment exerts a different influence compared to that resulting from the lack of the gene function.
人酪氨酰 - DNA磷酸二酯酶1（hTdp1）抑制剂NSC120686已与拓扑异构酶抑制剂一起用作药效团模型，以抑制Tdp1活性，作为癌症协同治疗的一部分。虽然该化合物在医学研究中有终端应用，但在植物中，其应用迄今尚未得到考虑。我们研究的独特之处在于在细胞中使用hTdp1抑制剂，这些细胞与人类细胞不同，含有两个基因。因此，本研究的目的是测试hTdp1抑制剂NSC120686作为一种探索工具来研究植物基因，因为它们的表征仍处于初始阶段。为此，将愈伤组织暴露于浓度不断增加（75、150、300μM）的NSC120686中。分别通过扩散试验和彗星试验测量的细胞死亡率和DNA损伤水平在使用最高剂量时显著增加，表明存在细胞毒性和遗传毒性阈值。此外，经NSC120686处理的愈伤组织和未处理的 - 缺失愈伤组织在程序性细胞死亡（PCD）/坏死和DNA损伤方面具有相似的反应。有趣的是，NSC120686处理对和基因的表达谱有不同影响，其中基因上调而基因下调。NSC120686处理不仅影响基因，还影响在替代DNA修复途径中起作用的其他基因。由于这些基因的表达模式与在 - 缺失植物中观察到的不同，因此可以假设NSC120686处理与缺乏基因功能所产生的影响相比具有不同的影响。
