Mechanisms Of Decitabine Induces G2/M Phase Arrest And Apoptosis In Kasumi-1 Cells

In recent years,DNA demethylation drugs have received more and more attention in the clinical treatment of hematological malignancies.However,high-efficiency and tumor cell-specific DNA demethylation drugs have not yet been discovered.Therefore,it is important to further reveal the role of DNA methylation modification in the occurrence and development of acute myeloid leukemia(AML)and the effect mechanism of Decitabine(DAC),but the specific regulatory mechanism Has not been revealed by the system.This study aims to further study the mechanism of DNA methyltransferase inhibitor DAC on AML cells and provide a theoretical basis for the treatment of leukemia.In order to clarify the expression of methyltransferase and DNA methylation in primary AML,this study first used the TCGA Cancer Genome Atlas database and DNA methylation modification to analyze,and found that the expression of DNMT3 a is up-regulated in human AML.Increased methylation modification.Next,DAC was added to culture the primary AML cell line Kasumi-1,and its biological behavior was tested.The results showed that DAC inhibited the proliferation of Kasumi-1 cells,affected the cell cycle arrest in G2/M phase,and promoted cell apoptosis.In order to further verify the regulation of DAC on the cell cycle,RT-q PCR was used to detect the expression of regulatory cell cycle regulators.The results showed that the expressions of p21,p53,CDC25 A,CDC2,CCNB1 and WEE1 were all up-regulated,indicating G2/M phase arrest The proliferation of Kasumi-1 cells may be inhibited by the change of p21/cdc25C/cdc2/cyclin B gene expression.In order to study the mechanism of the G2/M phase of Kasumi-1 cell cycle arrest caused by DAC treatment,firstly,using the TCGA database and online analysis of the UALCAN system,it was found that there were 541 genes negatively regulated with DNMT3 a in the cells of AML patients,among which calcium The binding protein S100A11 and DNMT3 a have a significant negative regulatory relationship,which is low in AML but not significant.Further transcriptome sequencing was performed on DAC-treated Kasumi-1 cells,and cross-analysis was performed on the genes up-regulated in the transcriptome sequencing and the AML in the TCGA and GEO databases with significantly lower methylation levels than normal bone marrow DNA.It was found that 204 genes were significantly changed in AML bone marrow and Kasumi-1 cells,including serine/threonine protein kinase ATM.Further verification found that the expression of ATM and P53 was significantly up-regulated after DAC treatment,indicating that DAC may promote the G2/M phase arrest and apoptosis of Kasumi-1cells by changing the expression of p53 and ATM.This study provides new ideas for the study of the mechanism of DNA methyltransferase in AML,and at the same time provides a new target for DAC intervention in the treatment of AML.