DataSheet_4_High expression level of the FTH1 gene is associated with poor prognosis in children with non-M3 acute myeloid leukemia.zip

Acute myelogenous leukemia (AML) is a disease that severely affects the physical health of children. Thus, we aimed to identify biomarkers associated with AML prognosis in children. Using transcriptomics on an mRNA dataset from 27 children with non-M3 AML, we selected genes from among those with the top 5000 median absolute deviation (MAD) values for subsequent analysis which showed that two modules were associated with AML risk groups. Thus, enrichment analysis was performed using genes from these modules. A one-way Cox analysis was performed on a dataset of 149 non-M3 AML patients downloaded from the TCGA. This identified four genes as significant: FTH1, RCC2, ABHD17B, and IRAK1. Through survival analysis, FTH1 was identified as a key gene associated with AML prognosis. We verified the proliferative and regulatory effects of ferroptosis on MOLM-13 and THP-1 cells using Liproxstatin-1 and Erastin respectively by CCK-8 and flow cytometry assays. Furthermore, we assayed expression levels of FTH1 in MOLM-13 and THP-1 cells after induction and inhibition of ferroptosis by real-time quantitative PCR, which showed that upregulated FTH1 expression promoted proliferation and inhibited apoptosis in leukemia cells. In conclusion, high expression of FTH1 promoted proliferation and inhibited apoptosis of leukemic cells through the ferroptosis pathway and is thus a potential risk factor that affects the prognosis of non-M3 AML in children.

急性髓系白血病（Acute myelogenous leukemia, AML）是一种严重危害儿童身体健康的疾病，本研究旨在鉴定与儿童AML预后相关的生物标志物。我们针对27例非M3型AML儿童的mRNA转录组数据集开展转录组学分析，选取中位绝对偏差（median absolute deviation, MAD）排名前5000的基因进行后续分析，结果发现两个基因模块与AML风险分组显著相关。遂对这两个模块中的基因进行富集分析。我们对从TCGA数据库下载的149例非M3型AML患者数据集进行单因素Cox回归分析，筛选出FTH1、RCC2、ABHD17B及IRAK1四个具有统计学显著性的基因。通过生存分析，确定FTH1为与AML预后相关的关键基因。我们分别使用Liproxstatin-1与Erastin处理MOLM-13及THP-1细胞，通过CCK-8实验与流式细胞术验证了铁死亡对这两种细胞的增殖与调控作用。此外，我们通过实时定量PCR检测了铁死亡诱导与抑制后MOLM-13及THP-1细胞中FTH1的表达水平，结果显示FTH1表达上调可促进白血病细胞增殖并抑制其凋亡。综上，FTH1高表达通过铁死亡通路促进白血病细胞增殖并抑制其凋亡，是影响儿童非M3型AML预后的潜在风险因素。