ZHOU Guizhen, PENG Lang, ZOU Lixin, LlU Xiaoliu

( Department of Hematology, the Fourth Hospital of Changsha, Changsha 410006, China)

AbstractObjective To investigate the effects of hypoxia on the proliferation and genomic methylation ofbone marrow mononuclear cells in patients with acute myeloid leukemia ( AML). Methods The bonemarrow fluid was collected from newly diagnosed AML patients, mononuclear cells were isolated. Cellswere 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 elfects of hypoxia onthe proliferation , metabolism , and apoptosis of bone marow mononuclear cells were analyzed using CCK-8 method , lactate dehydrogenase detection, and Hoechst 33258 staining. The 5-hydroxymethyleytosine ( 5.hmC ) and 5-methyleytosine ( 5-mC ) detection kits were used to detect the 5-hmC and 5-mC contents inthe genome of AML, cells , and the impact of hypoxia on the methylation status of the genome was analyzed.Results As the degree of hypoxia inereased, the proliferative activity and metabolic activity of AML cellswere inereased, while apoptosis was decreased , with a statistically significant differenee ( P<0.05). As thedegree of hypoxia inereased, the content of 5-hmC in the cell genome was increased, while the content of5-mC was decreased ( P<0.05 ). The proliferation and demethylation of cells were strongest when the cellswere cultured at a concentration of 1% 0, for 48 h. Conclusion Hypoxia promotes the proliferation andmetabolism of AMl, bone marrow mononuclear cells, inhibits cell apoptosis, and promotes cell genomedemethylation.

Keywords : hypoxia ; acute myeloid leukemia; methylation; 5-hydroxymethyleytosine ; 5-methyleytosine

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