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Whole-transcriptome sequencing (RNA-seq) analyses of the zebrafish liver cell line, ZFL, after acute exposure to Cu2+ ions.
Metallomics ( IF 2.9 ) Pub Date : 2020-03-13 , DOI: 10.1039/d0mt00005a Man Long Kwok 1 , Xue Lei Hu , Qi Meng , King Ming Chan
Metallomics ( IF 2.9 ) Pub Date : 2020-03-13 , DOI: 10.1039/d0mt00005a Man Long Kwok 1 , Xue Lei Hu , Qi Meng , King Ming Chan
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All cells require Cu as a cofactor, but Cu2+ induces toxicity and oxidative damage. A strict system is thus needed to maintain Cu homeostasis. Using the ZFL zebrafish liver cell line as a model, we studied the cellular responses after exposure to Cu2+, using whole-transcriptome shotgun sequencing (RNA-seq) to screen nearly all transcriptomes in cell samples and identify changes in gene expression. ZFL cells were treated with 100, 200, or 400 μM CuCl2 and harvested after 4 and 24 h. RNA was then extracted and subjected to RNA-Seq and qPCR validation. Exposure to 400 μM CuCl2 for 4 h and 24 h led to the regulation of 5993 and 4235 genes, respectively. In a gene ontology enrichment analysis, Cu2+ exposure enriched the nitrogen compound metabolic process and antioxidant activity but did not significantly affect cellular copper, zinc, iron and calcium ion homeostasis. In a KEGG pathway enrichment analysis, anti-oxidative stress induced the glutathione metabolism pathway. Furthermore, Cu2+ also induced genes related to apoptosis and arrested the cell cycle in the G2 phase. This study was based on the full gene expression profile combined with pathway analysis details, providing a full cellular response picture for Cu.
中文翻译：
急性暴露于Cu2 +离子后，斑马鱼肝细胞系ZFL的全转录组测序（RNA-seq）分析。
所有细胞都需要铜作为辅助因子，但是铜2+会诱导毒性和氧化损伤。因此需要一个严格的系统来维持Cu稳态。以ZFL斑马鱼肝细胞系为模型，我们研究了暴露于Cu 2+后的细胞反应，使用全转录组shot弹枪测序（RNA-seq）筛选了细胞样品中几乎所有的转录组并鉴定了基因表达的变化。ZFL细胞用100处理，200，或400μM的CuCl 2和后4和24小时收获细胞。然后提取RNA，并进行RNA-Seq和qPCR验证。暴露于400μMCuCl 2 4小时和24小时分别导致5993和4235基因的调控。在基因本体富集分析中，Cu 2+暴露丰富了氮化合物的代谢过程和抗氧化活性，但并未显着影响细胞铜，锌，铁和钙离子的体内稳态。在KEGG途径富集分析中，抗氧化应激诱导了谷胱甘肽代谢途径。此外，Cu 2+还诱导与凋亡相关的基因并使细胞周期停滞在G2期。这项研究基于完整的基因表达谱和途径分析的详细信息，为铜提供了完整的细胞反应图。
更新日期：2020-03-13
中文翻译：
急性暴露于Cu2 +离子后，斑马鱼肝细胞系ZFL的全转录组测序（RNA-seq）分析。
所有细胞都需要铜作为辅助因子，但是铜2+会诱导毒性和氧化损伤。因此需要一个严格的系统来维持Cu稳态。以ZFL斑马鱼肝细胞系为模型，我们研究了暴露于Cu 2+后的细胞反应，使用全转录组shot弹枪测序（RNA-seq）筛选了细胞样品中几乎所有的转录组并鉴定了基因表达的变化。ZFL细胞用100处理，200，或400μM的CuCl 2和后4和24小时收获细胞。然后提取RNA，并进行RNA-Seq和qPCR验证。暴露于400μMCuCl 2 4小时和24小时分别导致5993和4235基因的调控。在基因本体富集分析中，Cu 2+暴露丰富了氮化合物的代谢过程和抗氧化活性，但并未显着影响细胞铜，锌，铁和钙离子的体内稳态。在KEGG途径富集分析中，抗氧化应激诱导了谷胱甘肽代谢途径。此外，Cu 2+还诱导与凋亡相关的基因并使细胞周期停滞在G2期。这项研究基于完整的基因表达谱和途径分析的详细信息，为铜提供了完整的细胞反应图。
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