Investigation of the impact and mechanism of IRF2BP2 knockdown on the proliferation in acute myeloid leukemia cells
10.19405/j.cnki.issn1000-1492.2025.09.015
- VernacularTitle:敲除 IRF2BP2 对急性髓系白血病细胞增殖的影响及机制研究
- Author:
Bi Zhou
1
;
Xiaodong Tang
1
;
Ying Li
1
;
Yongping Zhang
2
;
Shaoyan Hu
2
Author Information
1. Dept of Pediatric，Suzhou Hospital of AnHui Medical University，Suzhou 234000
2. Dept of Hematology，Children’s Hospital of Soochow University，Suzhou 215000
- Publication Type:Journal Article
- Keywords:
acute myeloid leukemia;
IRF2BP2;
transcriptional regulation;
cell proliferation;
cell cycle;
CUT&Tag;
MYC
- From:
Acta Universitatis Medicinalis Anhui
2025;60(9):1682-1688
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To explore the effect of interferon regulatory factor 2 binding protein 2 ( IRF2BP2) on the proliferation of acute myeloid leukemia ( AML) cells and its molecular mechanism．
Methods:The CRISPR-Cas9 gene editing technology was used to knock out IRF2BP2 in human AML cell lines Kasumi-1 and U937，and West- ern blot was performed to detect the knockout efficiency of IRF2BP2 protein．Cell morphology was observed using a microscope．Cell phenotypes were analyzed by CCK-8 assay，colony formation experiments，and flow cytometry． RNA-Seq was performed to identify differentially expressed genes between the IRF2BP2 knockout group and the control group in the U937 cell line．Gene Set Enrichment Analysis ( GSEA) was conducted to explore the down- stream molecular mechanisms．Western blot was used to detect the expression of downstream differentially expressed genes．The Cleavage Under Targets and Tagmentation ( CUT＆Tag) technique was applied to identify the direct tar- gets of the IRF2BP2 protein，and the corresponding binding signals were visualized using the Integrated Genomics Viewer ( IGV) ．
Results:Compared with the control group，after knocking out IRF2BP2，the CCK-8 experiment showed that AML cell proliferation was inhibited ( P ＜0. 05) ; the number of colonies in the IRF2BP2 knockout group decreased ( P＜0. 05) ，and the proportion of G1 phase was prolonged ( P＜0. 05) ; in U937 cell lines，knoc- king out IRF2BP2 resulted in significant enrichment of differential genes in myelocytomatosis oncogene ( MYC) -re- lated signaling pathways，and the protein expression levels of pathway molecules MYC，cyclin-dependent kinase 4 ( CDK4) ，and cyclin － dependent kinase 2 ( CDK2 ) decreased with the downregulation of IRF2BP2; using IRF2BP2 antibodies in U937 cell lines for CUT＆Tag experiments，IGV visualization analysis showed a significant increase in signal peaks in the MYC promoter region．
Conclusion:IRF2BP2 protein affects the cell cycle and pro- liferation of AML cells by targeting and regulating MYC．
- Full text:2026032811544562294敲除IRF2BP2对急性髓系白血病细胞增殖的影响及机制研究_周碧.pdf
