Chen Yimin, Huang Zhenqian, Chen Shuyi, Tan Li, He Lang, Yuan Danyun, Zheng Lixia, Zhong Jing Hua, Li Anqiao, Zhang Heng, Tan Huo, Xu Lihua
Department of Hematology, The First Affiliated Hospital of Guangzhou Medical University, Guangzhou, 510230, China.
Department of Hematology, First People's Hospital of Foshan, 81 Linnan North Road, Chancheng, Foshan, Guangdong 528000, China.
Transl Oncol. 2023 Sep;35:101711. doi: 10.1016/j.tranon.2023.101711. Epub 2023 Jun 14.
Immediate early response 3 (IER3) plays a vital role in many tumors. This study aims to explore the function and mechanism of IER3 in Acute myeloid leukemia (AML).
The expression of IER3 in AML was performed by bioinformatics analysis. CCK-8 proliferation assay, flow cytometry cycle assay, clone formation assay, and tumorigenic ability were used to investigate the effect of IER3 on AML cells. Unbiased label-free quantitative proteomics and label-free quantitative phosphoproteomics analysis were performed. The regulatory relationship between SATB1(Special AT-rich sequence binding protein 1) and IER3 was investigated by Real time-PCR, Western blot, Chromatin immunoprecipitation (CHIP), and PCR.
The result indicated that the prognosis of the high IER3 expression group was significantly worse than that of the low expression group. CCK-8 assay showed that IER3 enhanced the proliferation ability. Cell cycle analysis showed IER3 could promote HL60 cells to enter the S phase of DNA synthesis from the quiescent phase. IER3 could stimulate HEL cells to enter mitosis. Clone-formation experiments suggested that IER3 enhanced clonogenic ability.IER3 promoted the tumorigenesis of AML. Further experimental investigation revealed that IER3 promoted autophagy and induced the occurrence and development of AML by negatively regulating the phosphorylation activation of AKT/mTOR pathway. SATB1 was found to bind to the promoter region of IER3 gene and negatively regulate its transcription.
IER3 could promote the development of AML and induce autophagy of AML cells by negatively regulating the phosphorylation and activation of AKT/mTOR. By the way, SATB1 may negatively target regulates IER3 transcription.
即刻早期反应3（IER3）在许多肿瘤中发挥着至关重要的作用。本研究旨在探讨IER3在急性髓系白血病（AML）中的功能及机制。
通过生物信息学分析检测AML中IER3的表达。采用CCK-8增殖试验、流式细胞术周期分析、克隆形成试验及致瘤能力研究IER3对AML细胞的影响。进行无偏倚的无标记定量蛋白质组学和无标记定量磷酸化蛋白质组学分析。通过实时PCR、蛋白质免疫印迹、染色质免疫沉淀（CHIP）和PCR研究特殊AT富集序列结合蛋白1（SATB1）与IER3之间的调控关系。
结果表明，IER3高表达组的预后明显差于低表达组。CCK-8试验表明IER3增强了增殖能力。细胞周期分析显示IER3可促进HL60细胞从静止期进入DNA合成的S期。IER3可刺激HEL细胞进入有丝分裂。克隆形成实验表明IER3增强了克隆形成能力。IER3促进了AML的肿瘤发生。进一步的实验研究表明，IER3通过负调控AKT/mTOR通路的磷酸化激活促进自噬并诱导AML的发生发展。发现SATB1与IER3基因的启动子区域结合并负调控其转录。
IER3可通过负调控AKT/mTOR的磷酸化和激活促进AML的发展并诱导AML细胞自噬。此外，SATB1可能负向靶向调控IER3转录。
