Effects of hypoxia on DNA hydroxymethylase Tet methylcytosine dioxygenase 2 in a KG‑1 human acute myeloid leukemia cell line and its mechanism

He,Ping; Lei,Jian; Zou,Li-Xin; Zhou,Gui-Zhen; Peng,Lang; Deng,Qian; Liu,Xiao-Liu

doi:10.3892/ol.2021.12953

Effects of hypoxia on DNA hydroxymethylase Tet methylcytosine dioxygenase 2 in a KG‑1 human acute myeloid leukemia cell line and its mechanism

Authors:
- Ping He
- Jian Lei
- Li-Xin Zou
- Gui-Zhen Zhou
- Lang Peng
- Qian Deng
- Xiao-Liu Liu
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Affiliations: Department of Hematology, The Affiliated Changsha Hospital, Hunan Normal University, Changsha, Hunan 410006, P.R. China, Department of Pathology, The Affiliated Tumor Hospital, Xiangya School of Medicine, Central South University, Changsha, Hunan 410013, P.R. China, Department of Hematology, Changsha Central Hospital, Changsha, Hunan 410018, P.R. China
Published online on: August 1, 2021 https://doi.org/10.3892/ol.2021.12953
Article Number: 692
Copyright: © He et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Hypoxia is involved in the epigenetic modification of leukemia. As an important DNA hydroxymethylase and a tumor suppressor gene, the expression regulating mechanism of Tet methylcytosine dioxygenase 2 (TET2) remains unclear. The aim of the present study was to explore whether hypoxia and hypoxia‑inducible factor 1α (HIF‑1α) regulate TET2 gene expression and its demethylation function in acute myeloid leukemia (AML). The human AML cell line KG‑1 was used in the present study. The results demonstrated that hypoxia could increase proliferation, enhance metabolism and inhibit apoptosis in KG‑1 cells, as detected by the cell counting kit‑8 assay, lactate dehydrogenase assay and Annexin V‑FITC/propidium iodide staining, respectively. Hypoxia reduced the genome methylation status in KG‑1 cells detected using 5‑methylcytosine and 5‑hydroxymethylcytosine detection kits. In addition, HIF‑1α overexpression increased TET2 expression, 5‑hmC level and cyclin‑dependent kinase inhibitor 2B [p15(INK4B)] gene demethylation compared with the HIF‑1α non‑overexpression group in KG‑1 cells detected by reverse transcription‑quantitative PCR, western blotting, 5‑hydroxymethylcytosine detection kits and methylation‑specific PCR, respectively. The inhibition of HIF‑1α by inhibitor YC‑1 reduced demethylation in KG‑1 cells by decreasing TET2 expression. It was also revealed that HIF‑1α could enhance TET2 transcriptional activity by binding to the hypoxia response element of the TET2 gene promoter region using chromatin immunoprecipitation and luciferase reporter gene assays. TET2 may be a potential target gene regulated by HIF‑1α. Hypoxia was demonstrated to regulate the expression of TET2 by HIF‑1α, which in turn affected the methylation and expression of downstream target genes and served a role in the occurrence and progression of leukemia. In the present study, the association between hypoxia metabolism and epigenetic regulation in AML was investigated and the findings provided a new idea and experimental basis for the diagnosis and treatment of hematologic malignancies.

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