Glycolytic metabolism confers resistance to combined all-trans retinoic acid and arsenic trioxide-induced apoptosis in HL60ρ0 cells
Introduction
Glycolytic cancers that do not rely on mitochondrial electron transport and oxidative phosphorylation for their energy requirements are characterized by high glucose intake [1], [2], [3], [4], [5], increased lactic acid production [2], [6], [7] and increased plasma membrane electron transport [8], [9], [10], [11]. In addition, glycolytic cancers are typically aggressive [3], [12], [13], [14] and often highly refractory to chemo- and radiation therapy [15], [16]. The effect of a glycolytic metabolism on cell processes such as differentiation and apoptosis has not been established. We have previously shown that a glycolytic metabolism confers resistance to neutrophilic differentiation in mitochondrial gene knockout mutants of the human leukemia cell line, HL60ρ0, after exposure to DMSO [17]. In this paper, we further investigate the effects of a glycolytic metabolism on the ability of HL60ρ0 cells to respond to the differentiation agents, ATRA and ATO alone or in combination.
ATRA combined with chemotherapy is the current standard of care for acute promyelocytic leukemia (APL), producing long lasting remissions in 75–85% of patients. Similarly, relapsed or refractory APL is often successfully treated with ATO [18], [19], [20], which causes partial differentiation and apoptosis of APL cells. Currently, several researchers are investigating the potential of ATRA and ATO as a combination therapy for relapsed and newly diagnosed patients with APL [21], [22], [23]. Although HL60 is not classified as an APL cell line as it does not have the t(15;17)(q22;q21) chromosomal translocation [24] and can differentiate along monocytic as well as granulocytic lineages [17], [25], it is capable of differentiating in response to ATRA and ATO. We therefore used HL60 and glycolytic HL60ρ0 cells to investigate the dependence of ATRA and ATO on mitochondrial respiration regarding their ability to induce differentiation and apoptosis.
Section snippets
Cells and cell culture
The mitochondrial DNA-knockout cell line, HL60ρ0, was derived from its parental cell line, HL60, by culturing in the presence of ethidium bromide for 6–8 weeks [26] and lack of mitochondrial DNA verified by PCR and stable phenotype. All cells were grown in RPMI-1640 medium (GIBCO-BRL, Grand Island, NY) supplemented with 5% (v/v) fetal bovine serum, glutamate (2 mM), penicillin (25 μg/ml), streptomycin (25 μg/ml), uridine (50 μg/ml) and pyruvate (1 mM) to densities of 1–2 × 106 cells/ml (exponential
Results
The effects of a glycolytic metabolism on the ability of ATRA and ATO to induce differentiation was investigated by exposing HL60 and HL60ρ0 cells to ATRA and ATO as single agents or in combination for up to 5 days.
We have previously reported an increase in expression of the neutrophil marker, CD15, on the surface of HL60 cells following 5 days exposure to DMSO, but only a marginal increase on HL60ρ0 cells [17]. Fig. 1A shows that ATRA and combined ATRA + ATO treatments caused a significant
Discussion
We have used the HL60/HL60ρ0 pair to investigate the effects of a purely glycolytic metabolism on the ability of ATRA and ATO alone or in combination to induce differentiation and cell death in leukemic cells. The ability of ATRA to induce differentiation of HL60 cells has been known for some time [30]. Here, we show that exposure to ATRA leads to differentiation in both HL60 and HL60ρ0 cells as determined by CD15 expression, albeit to a lesser extent in HL60ρ0 than HL60 cells, confirming and
Acknowledgments
This work was supported by the Cancer Society of New Zealand and the Radiation Therapy Department of the Wellington School of Medicine and Health Sciences, Otago University, New Zealand.
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