Transcription factor NFYA inhibits ferroptosis in lung adenocarcinoma cells by regulating PEBP1

IF 1.5 4区医学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Mutation Research-Fundamental and Molecular Mechanisms of Mutagenesis Pub Date : 2024-07-01 DOI:10.1016/j.mrfmmm.2024.111873

Feng Chen, Tingting Xu, Ni Jin, Digeng Li, Yanfu Ying, Chen Wang

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Abstract

Background

Ferroptosis is an iron-dependent programmed cell death mediated by lipid peroxidation. The purpose was to explore the molecular mechanism by which phosphatidylethanolamine-binding protein 1 (PEBP1) regulates ferroptosis in lung adenocarcinoma (LUAD), hoping to identify novel therapeutic targets for LUAD.

Methods

The expression, enrichment pathways and upstream transcription factors of PEBP1 were analyzed using bioinformatics tools. Dual-luciferase reporter assay and chromatin immunoprecipitation (ChIP) experiments were conducted to validate the interaction and binding relationship between PEBP1 and the upstream transcription factor nuclear transcription factor Y subunit α (NFYA). Quantitative reverse transcription PCR (qRT-PCR) was conducted to measure the expression levels of PEBP1 and NFYA mRNA in LUAD cells. Cell viability was detected by cell counting kit-8 assay. In addition, levels of malondialdehyde (MDA), Fe²⁺, and lipid reactive oxygen species (ROS) were assessed to evaluate ferroptosis levels in LUAD cells.

Results

PEBP1 was downregulated and significantly enriched in the ferroptosis signaling pathway in LUAD. Overexpression of PEBP1 suppressed cell viability remarkably, while levels of MDA, Fe²⁺, and lipid ROS were increased. Conversely, knockdown of PEBP1 produced the opposite effects. The upstream transcription factor NFYA, predicted to be involved in the regulation of PEBP1, was also upregulated in LUAD. Dual-luciferase reporter assay, ChIP, and molecular experiments revealed that NFYA transcriptionally suppressed the expression of PEBP1, and overexpression of NFYA could reverse the effects caused by PEBP1 overexpression.

Conclusion

PEBP1 regulated ferroptosis in LUAD, and the transcription factor NFYA inhibited ferroptosis in LUAD cells by transcriptionally downregulating PEBP1 expression.

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转录因子 NFYA 通过调控 PEBP1 抑制肺腺癌细胞的铁突变。

背景：铁变性是一种由脂质过氧化介导的铁依赖性程序性细胞死亡。本研究的目的是探讨磷脂酰乙醇胺结合蛋白1（PEBP1）调控肺腺癌（LUAD）铁凋亡的分子机制，希望能找到治疗LUAD的新靶点：方法：利用生物信息学工具分析了PEBP1的表达、富集途径和上游转录因子。双荧光素酶报告实验和染色质免疫沉淀（ChIP）实验验证了PEBP1与上游转录因子核转录因子Y亚基α（NFYA）之间的相互作用和结合关系。定量反转录 PCR（qRT-PCR）测定了 PEBP1 和 NFYA mRNA 在 LUAD 细胞中的表达水平。细胞活力通过细胞计数试剂盒-8检测。此外，还评估了丙二醛（MDA）、Fe2+和脂质活性氧（ROS）的水平，以评价 LUAD 细胞的铁变态反应水平：结果：在LUAD细胞中，PEBP1被下调，并在铁变态反应信号通路中显著富集。结果：在 LUAD 细胞中，PEBP1 被下调，并在铁氧化信号通路中明显富集。过表达 PEBP1 会明显抑制细胞活力，同时 MDA、Fe2+ 和脂质 ROS 水平升高。相反，敲除 PEBP1 则会产生相反的效果。上游转录因子 NFYA 被认为参与了 PEBP1 的调控，在 LUAD 中也被上调。双荧光素酶报告实验、ChIP和分子实验显示，NFYA转录抑制了PEBP1的表达，而NFYA的过表达可以逆转PEBP1过表达造成的影响：结论：PEBP1调控LUAD细胞的铁突变，转录因子NFYA通过转录下调PEBP1的表达抑制LUAD细胞的铁突变。

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来源期刊

Mutation Research-Fundamental and Molecular Mechanisms of Mutagenesis 生物-毒理学

CiteScore

4.90

自引率

0.00%

发文量

审稿时长

51 days

期刊介绍： Mutation Research (MR) provides a platform for publishing all aspects of DNA mutations and epimutations, from basic evolutionary aspects to translational applications in genetic and epigenetic diagnostics and therapy. Mutations are defined as all possible alterations in DNA sequence and sequence organization, from point mutations to genome structural variation, chromosomal aberrations and aneuploidy. Epimutations are defined as alterations in the epigenome, i.e., changes in DNA methylation, histone modification and small regulatory RNAs. MR publishes articles in the following areas: Of special interest are basic mechanisms through which DNA damage and mutations impact development and differentiation, stem cell biology and cell fate in general, including various forms of cell death and cellular senescence. The study of genome instability in human molecular epidemiology and in relation to complex phenotypes, such as human disease, is considered a growing area of importance. Mechanisms of (epi)mutation induction, for example, during DNA repair, replication or recombination; novel methods of (epi)mutation detection, with a focus on ultra-high-throughput sequencing. Landscape of somatic mutations and epimutations in cancer and aging. Role of de novo mutations in human disease and aging; mutations in population genomics. Interactions between mutations and epimutations. The role of epimutations in chromatin structure and function. Mitochondrial DNA mutations and their consequences in terms of human disease and aging. Novel ways to generate mutations and epimutations in cell lines and animal models.