Experimental study on treatment of relapsed and refractory acute myeloid leukemia with DNA methyltransferase 1 inhibitor combined with extracellular signal-regulated kinase 1,homeodomain-interacting protein kinase 2,and glycogen synthase kinase 3β inhibitors
- VernacularTitle:DNA甲基转移酶1抑制剂联合外胚层信号调节激酶1、同源结构域相互作用蛋白激酶2及糖原合成酶激酶3β治疗复发难治性急性髓系白血病的细胞实验研究
- Author:
Jingjing YU
1
;
Meng HU
;
ALIMU XIERENGULI
;
Man ZHANG
;
Jianhua QU
Author Information
- Keywords: inhibitors; DNA methyltransferase 1; acute myeloid leukemia; extracellular sig-nal-regulated kinase 1; homeodomain-interacting protein kinase 2; glycogen synthase kinase 3 β; targeted therapy; cell apoptosis; protein arrest
- From: Journal of Clinical Medicine in Practice 2025;29(5):26-30,35
- CountryChina
- Language:Chinese
- Abstract: Objective To investigate the mechanism of DNA methyltransferase 1(DNMT1)inhibitor combined with extracellular signal-regulated kinase 1(ERK1),homeodomain-interacting protein kinase 2(HIPK2),and glycogen synthase kinase 3 β(GSK3 β)inhibitors in synergistically inducing apoptosis and protein arrest in relapsed and refractory acute myeloid leukemia(AML)cells.Methods Three therapeutic targets,including ERK1,HIPK2,and GSK3β,were screened based on the Gene Expression Omnibus(GEO),The Cancer Genome Atlas(TCGA)database,and Ex-pression 2 Kinases database.Human AML cell line U937 cells were treated with DNMT1 inhibitor a-lone or combined with ERK1,HIPK2,or GSK3β inhibitors.Cell viability was detected using the CCK-8 method.Apoptosis rate was analyzed by flow cytometry,and cell cycle distribution was de-termined by propidium iodide(PI)staining.The mRNA expression levels of DNMT1,ERK1,HIPK2,and GSK3β were detected by real-time fluorescent quantitative reverse transcriptionpoly-merase chain reaction(RT-qPCR).Protein expressionlevels of DNMT1,ERK1,HIPK2,and GSK3β were detected by immunoblotting.Results DNMT1 inhibitor significantly inhibited the cell viability of U937 cells,and significantly induced apoptosis and cell cycle arrest in U937 cells(P＜0.05).When DNMT1 inhibitor was combined with ERK1,HIPK2,or GSK3β inhibitors,cell via-bility and apoptosis rate were significantly reduced(P＜0.05).DNMT1 inhibitor alone or its com-bination with ERK1,HIPK2,and GSK3β inhibitors induced U937 cell arrest in the G0/G,phase,with a significant increase in the proportion of cells in the G0/G1 phase in the combination group(P＜0.05).The combination of DNMT1 inhibitor with ERK1,HIPK2,and GSK3β inhibitors sig-nificantly reduced the mRNA expression levels in DNMT1,ERK1,HIPK2,and GSK3β in U937 cells(P＜0.05).Similarly,the combination therapy significantly reduced the protein expression levels of DNMT1,ERK1,HIPK2,and GSK3β in U937 cells(P＜0.05).Conclusion DNMT1 inhibitor combined with ERK1,HIPK2,and GSK3 β inhibitors can synergistically induce apoptosis and protein arrest in relapsed and refractory AML cells,providing a novel strategy for combined tar-geted therapy of AML.
