Microcystin-LR-regulated transcriptome dynamics in ZFL cells
Microcystin-LR-regulated transcriptome dynamics in ZFL cells
复制标题
ZFL 细胞中微囊藻毒素-LR 调节的转录组动力学
DOI：
10.1016/j.aquatox.2019.04.018
复制
发表时间：
2019-07-01
影响因子：
4.5
通讯作者：
Zhong, Shan
中科院分区：
文献类型：
--
作者：
Lu, Xing;Tian, Juan;Zhong, Shan
微囊藻毒素-LR（MC-LR）是一种剧毒肝毒素，对水生生物乃至人类健康造成极大危害。斑马鱼肝细胞系（ZFL）是研究毒物毒性和代谢的有价值的模型。然而，MC-LR 的毒性及其对 ZFL 细胞基因转录的影响仍有待表征。在本研究中，我们确定了 MC-LR 对 ZFL 细胞的毒性，并研究了 MC-LR 对细胞转录组动力学的影响。 MC-LR对ZFL细胞的EC50为80.123μg/mL。将ZFL细胞暴露于10μg/mL MC-LR 0、1、3、6、12或24小时，并进行RNA测序以分析基因转录。总共发现 10,209 个基因受 MC-LR 调节。不同时间点上调和下调基因的数量分别为2179至3202和1501至2597。此外，1543个基因在MC-LR暴露后经历了差异剪接（AS），其中620个未被识别为差异表达基因（DEG）。 MC-LR 对细胞功能的影响高度依赖于时间。 MAPK（丝裂原激活蛋白激酶）和 FoxO（叉头盒 O）信号通路是 MC-LR 激活的最重要的通路。类固醇生物合成和萜类主链生物合成是下调基因最丰富的。根据表达谱数据集构建了基因调控网络，并确定了候选主转录因子。我们的结果揭示了 MC-LR 细胞毒性的分子机制以及 MC-LR 毒性后 ZFL 细胞的转录组图谱。
Microcystin-LR (MC-LR) is a highly toxic hepatotoxin that poses great hazards to aquatic organisms and even human health. The zebrafish liver cell line (ZFL) is a valuable model for investigating toxicity and metabolism of toxicants. However, the toxicity of MC-LR and its effects on gene transcription of ZFL cells remains to be characterized. In this study, we determined the toxicity of MC-LR for ZFL cells and investigated the effects of MC-LR on cellular transcriptome dynamics. The EC50 of MC-LR for ZFL cells was 80.123 mu g/mL. The ZFL cells were exposed to 10 mu g/mL MC-LR for 0, 1, 3, 6, 12 or 24 h, and RNA-sequencing was performed to analyze gene transcription. A total of 10,209 genes were found to be regulated by MC-LR. The numbers of up- and down-regulated genes at different time points ranged from 2179 to 3202 and from 1501 to 2597, respectively. Furthermore, 1543 genes underwent differential splicing (AS) upon MC-LR exposure, of which 620 were not identified as differentially expressed gene (DEG). The effects of MC-LR on cellular functions were highly time-dependent. MAPK (mitogen-activated protein kinase) and FoxO (forkhead box O) signaling pathways were the most prominent pathways activated by MC-LR. Steroid biosynthesis and terpenoid backbone biosynthesis were the most enriched for the down-regulated genes. A gene regulatory network was constructed from the expression profile datasets and the candidate master transcription factors were identified. Our results shed light on the molecular mechanisms of MC-LR cellular toxicity and the transcriptome landscapes of ZFL cells upon MC-LR toxicity.