Effects of ferroptosis-related gene HSPB1 on acute myeloid leukemia

IF 2.2 4区医学 Q3 HEMATOLOGY

International Journal of Laboratory Hematology Pub Date : 2024-06-02 DOI:10.1111/ijlh.14319

Xue-Shen Yan, Yu-Jiao Sun, Juan Du, Wen-Yan Niu, Han Qiao, Xiang-Cong Yin

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Abstract

Introduction

The purpose of this study was to investigate the effects and potential mechanisms of ferroptosis-related gene heat shock protein beta-1 (HSPB1) on acute myeloid leukemia (AML).

Methods

The RNA-seq and clinical data of AML samples were obtained from the Genomic Data Commons database, and the FerrDb database was used to screen the marker, drive and suppressor of ferroptosis. Besides, DESeq2 was applied for differential expression analysis on AML samples and screening for differentially expressed genes (DEGs). The screened DEGs were subjected to the intersection analysis with ferroptosis-related genes to identify the ferroptosis-related DEGs. Next, the functional pathways of ferroptosis-related DEGs were further be discussed by Gene Ontology as well as Kyoto Encyclopedia of Genes and Genomes enrichment analysis of DEGs. Additionally, lasso regression analysis was employed to determine the differential genes related to prognosis in patients with AML and the survival analysis was performed. Subsequently, quantitative real-time polymerase chain reaction and western blot assay were applied to detect the mRNA and protein expression levels of HSPB1 in normal/AML bone marrow tissues and human normal (HS-5)/AML (HL-60) bone marrow cells, respectively. Furthermore, HSPB1 was knocked down to assess the expression changes of glutathione peroxidase 4 and acyl-CoA synthetase long-chain family member 4. Ultimately, the viability and oxidative stress levels of HL-60 were analyzed by Cell Counting Kit-8 and biochemical detection.

Results

A total of 4986 DEGs were identified in AML samples, with 3324 up-regulated and 1662 down-regulated. The enrichment analysis illustrated that ferroptosis-related DEGs were significantly enriched in response to metal irons, oxidative stress, and other pathways. After lasso regression analysis, 17 feature genes related to the prognosis of patients with AML were obtained, with HSPB1 exhibiting a significant correlation. The reliability of our models was verified by Cox regression analysis and survival analysis of the hazard model. Furthermore, the outcomes of quantitative real-time polymerase chain reaction and western blot showed that mRNA and protein expression levels of HSPB1 were significantly increased in the AML Group and HL-60 cells. The knockdown of HSPB1 in HL-60 cells reduced the protein level of glutathione peroxidase 4, increased the protein level of acyl-CoA synthetase long-chain family member 4, decreased the cell viability, and aggravated oxidative stress.

Conclusion

Ferroptosis-related gene HSPB1 is highly expressed in patients with AML. In addition, HSPB1 may be involved in the occurrence and development of AML by regulating oxidative stress and ferroptosis-related pathways. This study provides new clues for further understanding of AML molecular mechanisms. Also, HSPB1 is expected to be a potential therapeutic target for AML in the future.

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铁突变相关基因 HSPB1 对急性髓性白血病的影响

引言本研究旨在探讨铁突变相关基因热休克蛋白β-1（HSPB1）对急性髓性白血病（AML）的影响及潜在机制：方法：从Genomic Data Commons数据库获取AML样本的RNA-seq和临床数据，并利用FerrDb数据库筛选铁突变的标记物、驱动因子和抑制因子。此外，还应用 DESeq2 对 AML 样本进行差异表达分析，筛选差异表达基因（DEGs）。筛选出的 DEGs 与铁变态反应相关基因进行交叉分析，以确定与铁变态反应相关的 DEGs。接下来，通过基因本体论以及京都基因和基因组百科全书对 DEGs 的富集分析，进一步探讨了铁变态反应相关 DEGs 的功能通路。此外，还利用拉索回归分析确定了与急性髓细胞性白血病患者预后相关的差异基因，并进行了生存分析。随后，应用实时定量聚合酶链反应和免疫印迹法分别检测了HSPB1在正常/AML骨髓组织和人正常（HS-5）/AML（HL-60）骨髓细胞中的mRNA和蛋白表达水平。此外，通过敲除 HSPB1 来评估谷胱甘肽过氧化物酶 4 和酰基-CoA 合成酶长链家族成员 4 的表达变化。最后，通过细胞计数试剂盒-8和生化检测分析了HL-60的活力和氧化应激水平：结果：在急性髓细胞样本中共发现了 4986 个 DEGs，其中 3324 个上调，1662 个下调。富集分析表明，与铁突变相关的 DEGs 在金属铁、氧化应激和其他通路中均有显著富集。经过套索回归分析，得到了 17 个与急性髓细胞性白血病患者预后相关的特征基因，其中 HSPB1 表现出明显的相关性。Cox回归分析和危险模型生存分析验证了我们的模型的可靠性。此外，实时定量聚合酶链反应和免疫印迹的结果表明，HSPB1的mRNA和蛋白表达水平在AML组和HL-60细胞中明显升高。HSPB1在HL-60细胞中的敲除降低了谷胱甘肽过氧化物酶4的蛋白水平，增加了酰基-CoA合成酶长链家族成员4的蛋白水平，降低了细胞活力，加重了氧化应激：结论：铁突变相关基因HSPB1在急性髓细胞性白血病患者中高表达。结论：铁蛋白沉积相关基因HSPB1在急性髓细胞性白血病患者中高表达，而且HSPB1可能通过调节氧化应激和铁蛋白沉积相关途径参与急性髓细胞性白血病的发生和发展。这项研究为进一步了解急性髓细胞性白血病的分子机制提供了新的线索。同时，HSPB1有望成为未来治疗急性髓细胞性白血病的潜在靶点。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

International Journal of Laboratory Hematology 医学-血液学

CiteScore

4.50

自引率

6.70%

发文量

211

审稿时长

6-12 weeks

期刊介绍： The International Journal of Laboratory Hematology provides a forum for the communication of new developments, research topics and the practice of laboratory haematology. The journal publishes invited reviews, full length original articles, and correspondence. The International Journal of Laboratory Hematology is the official journal of the International Society for Laboratory Hematology, which addresses the following sub-disciplines: cellular analysis, flow cytometry, haemostasis and thrombosis, molecular diagnostics, haematology informatics, haemoglobinopathies, point of care testing, standards and guidelines. The journal was launched in 2006 as the successor to Clinical and Laboratory Hematology, which was first published in 1979. An active and positive editorial policy ensures that work of a high scientific standard is reported, in order to bridge the gap between practical and academic aspects of laboratory haematology.