Glycolytic metabolism confers resistance to combined all-trans retinoic acid and arsenic trioxide-induced apoptosis in HL60ρ0 cells

doi:10.1016/j.leukres.2007.04.014

Leukemia Research

Volume 32, Issue 2, February 2008, Pages 327-333

https://doi.org/10.1016/j.leukres.2007.04.014 Get rights and content

Abstract

Glycolytic cancers are resistant to many forms of chemotherapy and some respond poorly to differentiation therapies. Here, we investigate the effects of exposure to all-trans retinoic acid (ATRA) and arsenic trioxide (ATO) on differentiation and cell survival in the human leukemia cell line, HL60 and its mitochondrial gene knockout mutant, HL60ρ⁰. Glycolytic HL60ρ⁰ cells exposed to single and combined treatments expressed less CD15, in most cases, but produced a stronger respiratory burst than parental HL60 cells. HL60ρ⁰ cells were also significantly more resistant to apoptosis after combined ATO + ATRA treatment compared with HL60 cells, and this was associated with failure to upregulate Fas expression.

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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View full text

Glycolytic metabolism confers resistance to combined all-trans retinoic acid and arsenic trioxide-induced apoptosis in HL60ρ0 cells

Abstract

Introduction

Section snippets

Cells and cell culture

Results

Discussion

Acknowledgments

Neoplasia

Cancer Cell

J Biol Chem

Biochim Biophys Acta

Biochim Biophys Acta

Neoplasia

J Biol Chem

Leuk Res

Blood

Blood

Blood

Cancer Gen Cytogen

J Immunol Methods

Blood

Ist die Glykolyse spezifisch für die Tumoren?

Biochem Z

Tumor mitochondria and the bioenergetics of cancer cells

Prog Exp Tumor Res

Overexpression of Glut-1 and increased glucose metabolism in tumors are associated with a poor prognosis in patients with oral squamous cell carcinoma

Cancer

Expression of GLUT-1 glucose transfer, cellular proliferation and grade of tumour correlate with [F-18]-fluorodeoxyglucose uptake by positron emission tomography in epithelial tumors of the ovary

Int J Cancer

Glycolytic metabolism confers resistance to combined all-trans retinoic acid and arsenic trioxide-induced apoptosis in HL60ρ⁰ cells