Adaptation of leukemia cells to hypoxic condition through switching the energy metabolism or avoiding the oxidative stress

BMC Cancer

Masakazu Nitta¹  Ichiro Hanamura¹  Shohei Mizuno¹  Motonori Mizutani¹  Masato Shikami²  Norikazu Tsunekawa-Imai¹  Kazuto Suganuma¹  Hiroshi Miwa¹  Mineaki Goto¹ 

[1]Department of Internal Medicine, Division of Hematology, Aichi Medical University School of Medicine, 1-1 Yazakokarimata, Nagakute, Aichi 480-1195, Japan

[2]Department of Internal Medicine, Division of Hematology, Daiyukai General Hospital, Ichinomiya, Aichi 491-0113, Japan

关键词: Cytochrome c oxidase;    Pyruvate dehydrogenase kinase;    Reactive oxygen species;    Energy metabolism;    Hypoxia;    Leukemia;   

Others  :  859081
DOI  :  10.1186/1471-2407-14-76

 received in 2013-07-29, accepted in 2014-02-06,  发布年份 2014

【 摘 要 】

Background

Like normal hematopoietic stem cells, leukemia cells proliferate in bone marrow, where oxygen supply is limited. However, the growth and energy metabolism of leukemia cells under hypoxia have not been well understood. Although it has been known that reactive oxygen species (ROS) is generated under hypoxic conditions, normal and leukemia stem cells were characterized by relatively low levels of ROS. Roles of ROS on leukemia cells under hypoxia also have not been well understood.

Methods

Four Leukemia cell lines were cultured under normoxia (21% O₂) or hypoxia (1% O₂), where NB4 and THP-1 were most extensively studied. To evaluate energy metabolism, we estimated whole cell number or apoptotic cells with or without a glycolysis inhibitor or an oxidative phosphorylation (OXPHOS) inhibitor. Glucose consumption and lactate production were also measured. To evaluate oxidative stress in hypoxic condition, the ROS level and GSH (reduced glutathione) / GSSG (oxidized glutathione) ratio was measured. In addition, pyruvate dehydrogenase kinase 1 (PDK1) and cytochrome c oxidase subunit 4 (COX4) were examined by western blotting or RT-PCR.

Results

NB4, which grows well under normoxia depending on glycolysis, demonstrated prominent apoptosis and growth suppression after 48 hours culture under hypoxia. NB4 cells cultured under hypoxia showed significantly increased ROS. Culture with a ROS scavenger resulted in decrease of apoptotic cell death of NB4 under hypoxia. NB4 cells cultured for longer period (7 days) under hypoxia did not come to extinction, but grew slowly by upregulating GSH synthesis to protect from ROS generated in hypoxic condition. By contrast, THP-1, which largely depends on OXPHOS in mitochondria under normoxia, demonstrated more growth under hypoxia by changing metabolism from OXPHOS to glycolysis through upregulating PDK1. Moreover, THP-1 avoided ROS generation by substituting COX 4 subunit (from COX 4–1 to COX 4–2) through upregulation of LON, a mitochondrial protease under hypoxia.

Conclusions

We showed that leukemia cells survive and adapt to the hypoxic condition through various pathways. Our results will help understanding energy metabolism of leukemia cells and creating novel therapeutics.

【 授权许可】

   
2014 Goto et al.; licensee BioMed Central Ltd.

【 图 表 】

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