抗肿瘤药物FTY720对p53依赖的基因转录调控

Anti-tumor drug FTY720 regulates p53-dependent transcription

目的探究抗肿瘤药物FTY720是否参与p53依赖的基因表达调控,分析FTY720-p53轴潜在的生物学调控作用。方法在表达野生型p53的人骨肉瘤细胞系U2OS中利用CRISPR/Cas9技术构建p53敲除(p53-KO)的细胞株;将野生型或p53-KO的U2OS细胞给予FTY720处理,通过转录组测序(RNA-seq)结合基因功能富集分析表征p53依赖的差异表达基因及其潜在的生物学功能。结果获得U2OS p53-KO细胞株,筛选出220个FTY720作用下p53依赖的差异表达基因。这些基因主要在血小板活化、细胞趋化因子反应、Rho蛋白信号传导、多细胞生物体内平衡、黏蛋白型O-聚糖生物合成、细胞外基质(ECM)-受体相互作用、病毒蛋白质与细胞因子和细胞因子受体相互作用等生物学过程中起着调控作用。结论p53的激活可能在FTY720抑制肿瘤进展的过程中起到一定作用。

Objective To explore whether anti-tumor drug FTY720 participates in regulation of p53-dependent gene transcription and to evaluate the potential roles of FTY720-p53 axis in the modulation of cellular biological processes.Methods CRISPR-Cas9 technique was used to generate a p53-knockout(p53-KO)cell strain with human osteosarcoma cell line U2OS that expressed the wildtype p53.Control cells or p53-KO cells were treated with or without FTY720 and the transcriptional was then assessed by RNA-sequencing(RNA-seq)analysis.The differentially expressed genes as well as their potential functions were annotated through bioinformatic analysis.Results The U2OS p53-KO cell strain was successfully established.220 p53-dependent downstream genes were identified in response to FTY720 treatment.These genes were functionally enriched in the regulation of multiple biological processes including negative regulation of platelet activation,cellular response to chemokine,positive regulation of Rho protein signal transduction,multicellular organismal homeostasis,mucin type O-glycan biosynthesis,ECM-receptor interaction,viral protein interaction with cytokine and cytokine receptor.Conclusions p53 activation is involved in FTY720-mediated tumor suppressive functions.