Hydrogen peroxide (H2O2) induces leukemic but not normal hematopoietic cell death in a dose-dependent manner

Amanda Nogueira-Pedro,Carolina Carvalho Dias,Edgar Julian Paredes-Gamero,Clarice Silvia Taemi Origassa,Lilian Piñero Marcolin Eça,Thalyta Aparecida Munhoz Cesário,Alice Teixeira Ferreira

doi:10.1186/1475-2867-13-123

Amanda Nogueira-Pedro, Carolina Carvalho Dias + Show 5 more

Open Access

https://doi.org/10.1186/1475-2867-13-123

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Abstract

Over the last few years, studies have suggested that oxidative stress plays a role in the regulation of hematopoietic cell homeostasis. In particular, the effects of hydrogen peroxide (H2O2) range from hematopoietic cell proliferation to cell death, depending on its concentration in the intracellular milieu. In this work, we evaluated the effects of an oxidative environment on normal and leukemic hematopoietic cells by stimulating normal human (umbilical cord blood) and murine (bone marrow) hematopoietic cells, as well as human myeloid leukemic cells (HL-60 lineage), upon H2O2 stimulus. Total cell populations and primitive subsets were evaluated for each cell type. H2O2 stimulus induces HL-60 cell death, whereas the viability of human and murine normal cells was not affected. The effects of H2O2 stimulus on hematopoietic stem/progenitor cell subsets were examined and the normal primitive cells were found to be unaffected; however, the percentage of leukemic stem cells (LSC) increased in response to H2O2, while clonogenic ability of these cells to generate myeloid clones was inhibited. In addition, H2O2 stimulus caused a decrease in the levels of p-AKT in HL-60 cells, which most likely mediates the observed decrease of viability. In summary, we found that at low concentrations, H2O2 preferentially affects both the LSC subset and total HL-60 cells without damage normal cells.

Highlights

Hematopoiesis is a continuous process that occurs throughout the life of an organism in which blood cells are produced in an organized and hierarchical process of development initiated in the bone marrow by hematopoietic stem cells (HSC) [1]
Murine normal hematopoietic cell extraction and culture The femurs from three-month-old male C57BL/6 mice were extracted, and bone marrow cells were collected in laminar flux by flushing the cells with Iscove’s Modified Dulbecco’s Medium (IMDM) supplemented with 12.5% fetal bovine serum (FBS), 12.5% fetal calf serum (FCS) and 1% antibiotics
HL-60 cell viability is targeted by H2O2 Initially, the cytotoxicity of H2O2 in different cell types was tested, and dose–response curves were built

Summary

Introduction

Hematopoiesis is a continuous process that occurs throughout the life of an organism in which blood cells are produced in an organized and hierarchical process of development initiated in the bone marrow by hematopoietic stem cells (HSC) [1]. Decomposition of hydrogen peroxide (H2O2) by catalase increases granulocytes and HSC number in long-term bone marrow cultures [6]. It was shown the existence of two distinct HSC population based on their ROS content, where HSC ROSlow population, but not HSC ROShigh, exhibits the characteristics of a typical HSC [7]. ROS generation is important because its accumulation results in hemolysis and shortened red blood cell lifespan [11,12]. These and other reports demonstrate the importance of ROS in the biology of HSC

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