缺氧对AML骨髓单个核细胞的增殖及基因组甲基化的影响  

Effects of hypoxia on the proliferation and genomic methylation of acute myeloid leukemia bone marrow mononuclear cells

作　　者：周贵珍 彭浪[1] 邹立新[1] 刘小柳[1] ZHOU Guizhen;PENG Lang;ZOU Lixin;LIU Xiaoliu(Department of Hematology,the Fourth Hospital of Changsha,Changsha 410006,China)

机构地区：[1]长沙市第四医院血液肿瘤科,长沙410006

出　　处：《华夏医学》2024年第1期44-49,共6页Acta Medicinae Sinica

基　　金：国家自然科学基金项目(81500141);湖南省卫健委课题(202303046746)。

摘　　要：目的研究缺氧对急性髓系白血病(AML)患者骨髓单个核细胞的增殖及基因组甲基化的影响。方法收集初诊AML患者的骨髓液,分离单个核细胞,以不同浓度的氧(氧含量分别为21%、3%和1%)处理细胞24、48、72 h,以常氧(氧含量21%)作为对照。CCK-8法、乳酸脱氢酶检测及Hoechst 33258染色等分析缺氧对骨髓单个核细胞的增殖、代谢及凋亡的影响。采用5-羟甲基化胞嘧啶(5-hmC)和5-甲基化胞嘧啶(5-mC)检测试剂盒分析缺氧对基因组甲基化状态的影响。结果随着缺氧程度的增加,AML细胞增殖活性增强,代谢活性增加,而细胞凋亡减少,差异有统计学意义(P<0.05)。随着缺氧程度增加,细胞基因组5-hmC的含量升高,5-mC的含量降低(P<0.05)。浓度1%的O_(2)培养细胞48 h,细胞的增殖和去甲基化最强。结论缺氧促进AML患者骨髓单个核细胞的增殖与代谢,抑制细胞凋亡,促进细胞基因组去甲基化。Objective To investigate the effects of hypoxia on the proliferation and genomic methylation of bone marrow mononuclear cells in patients with acute myeloid leukemia(AML).Methods The bone marrow fluid was collected from newly diagnosed AML patients,mononuclear cells were isolated.Cells were treated with different concentrations of oxygen(oxygen content 21%,3%,and 1%,respectively)for 24,48,and 72 h,with normal oxygen(oxygen content 21%)as the control.The effects of hypoxia on the proliferation,metabolism,and apoptosis of bone marrow mononuclear cells were analyzed using CCK-8 method,lactate dehydrogenase detection,and Hoechst 33258 staining.The 5-hydroxymethylcytosine(5-hmC)and 5-methylcytosine(5-mC)detection kits were used to detect the 5-hmC and 5-mC contents in the genome of AML cells,and the impact of hypoxia on the methylation status of the genome was analyzed.Results As the degree of hypoxia increased,the proliferative activity and metabolic activity of AML cells were increased,while apoptosis was decreased,with a statistically significant difference(P<0.05).As the degree of hypoxia increased,the content of 5-hmC in the cell genome was increased,while the content of 5-mC was decreased(P<0.05).The proliferation and demethylation of cells were strongest when the cells were cultured at a concentration of 1%O_(2) for 48 h.Conclusion Hypoxia promotes the proliferation and metabolism of AML bone marrow mononuclear cells,inhibits cell apoptosis,and promotes cell genome demethylation.

关 键 词：缺氧 急性髓系白血病 甲基化 5-羟甲基化胞嘧啶 5-甲基化胞嘧啶 

分 类 号：R557[医药卫生—血液循环系统疾病]