白藜芦醇通过Hsa-miR-335-5p/NFS1/GPX4通路以ROS依赖的方式诱导急性髓系白血病细胞的铁死亡。
Resveratrol drives ferroptosis of acute myeloid leukemia cells through Hsa-miR-335-5p/NFS1/ GPX4 pathway in a ROS-dependent manner.
发表日期:2023 Jul 31
作者:
Jia Liu, Wei Gao, Yong Sheng, Jiafeng Sun, Donghu Wen
来源:
GENES & DEVELOPMENT
摘要:
探索诱导铁死亡治疗急性髓系白血病(AML)的潜在靶点及其机制与潜在药物。使用关键词“急性髓系白血病”,在TCGA和GEO数据库中搜索差异表达基因。通过ROC曲线、AUC值和生存分析进行筛选,再通过RT-qPCR和Western-blot分析验证AML-193和OCI-AML-3细胞中NFS1的高表达水平。在进行CCK-8检测时加入各种细胞死亡抑制剂与不加抑制剂对比,通过GPX4的表达水平进一步检测铁死亡。在Starbase和TargetScan中找到了NFS1的上游调控miRNA。然后通过在AML-193和OCI-AML-3细胞中进行沉默和上调实验来确定hsa-miR-335-5p、NFS1和GPX4之间的关系。此外,通过DCFH-DA检测细胞ROS的含量。最后,使用白藜芦醇增强AML-193和OCI-AML-3细胞中的铁死亡。NFS1在AML细胞中高表达,与AML相关的死亡率呈正相关,可用于诊断AML。敲除NFS1促进ROS积累和与铁死亡相关的易变铁库增加。si-NFS1可抑制GPX4的表达水平,促进ROS积累并诱导与铁死亡相关的易变铁库增加。此外,在si-NFS1处理后,过度表达的GPX4可导致细胞死亡水平下调。hsa-miR-335-5p被发现为NFS1的上游调控因子。通过sh-hsa-miR-335-5p转染可上调NFS1的表达,并通过hsa-miR-335-5p转染可抑制NFS1的表达。白藜芦醇可增加hsa-miR-335-5p的表达水平,降低NFS1和GPX4的表达。白藜芦醇可通过ROS依赖途径增强AML细胞的铁死亡,作用机制为Hsa-miR-335-5p/NFS1/GPX4信号通路。
To explore the potential target to induce ferroptosis for treating acute myeloid leukemia (AML) as well as its mechanism and latent drugs. Using the keyword "acute myelogenous leukemia", the related dataset in TCGA and GEO were used for searching differentially expressed genes. After the filtrate by ROC curve, AUC values, and survival analysis, RT-qPCR as well as Western-blot analysis were performed to verify the high expression level of NFS1 in AML-193 and OCI-AML-3 cells. After CCK-8 detection with and without various cell death inhibitors, ferroptosis were further detected by the expression level of GPX4. After taking the intersection in Starbase and TargetScan, the upstream regulatory miRNA of NFS1 was found. Then the relation of hsa-miR-335-5p, NFS1, as well as GPX4, was ascertained by knockdown and overexpression study in AML-193 and OCI-AML-3 cells. In addition, cellular ROS was detected by DCFH-DA. Finally, resveratrol was used to intensify ferroptosis of AML-193 and OCI-AML-3 cells. NFS1 was highly expressed in AML cells, positively associated with AML-related mortality, and can be used to diagnose AML. Knockout of NFS1 facilitated ROS accumulation and ferroptosis-associated labile iron pool increase. si-NFS1 can inhibit the expression level of GPX4, facilitate ROS accumulation and induce ferroptosis-associated labile iron pool increase. Besides, overexpressed GPX4 can lead to down-regulated cell death after si-NFS1 treatment. Hsa-miR-335-5p was found as the upstream regulator of NFS1. The expression of NFS1 can be up-regulated by sh-hsa-miR-335-5p transfection and can be inhibited by hsa-miR-335-5p transfection. Resveratrol was found can increase the expression level of hsa-miR-335-5p and decrease the expression of NFS1 and GPX4. Resveratrol can intensify ferroptosis of AML cells via Hsa-miR-335-5p/NFS1/ GPX4 pathway through a ROS-dependent manner.