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The Antiproliferative Effect of EPA in HL60 Cells is Mediated by Alterations in Calcium Homeostasis

  • Original Article
  • Published:
Lipids

Abstract

Studies show that n-3 polyunsaturated fatty acids (PUFA) inhibit proliferation and induce apoptosis in cancer cells. Recent reports indicate that this effect is due to activation of the unfolded protein response (UPR). However, what causes this activation has been unclear. We examined the effects of eicosapentaenoic acid (EPA) on the human leukemia cell line HL60 and the econazole (Ec) resistant HL60 clone E2R2. Ec depletes Ca2+ from the ER and blocks Ca2+ influx in mammalian cells, leading to activation of the UPR and apoptosis. EPA inhibited growth of HL60 cells strongly, while E2R2 cells were much less affected. Gene expression analysis of HL60 cells revealed extensive changes in transcripts related to the ER homeostasis, Ca2+-homeostasis and cell cycle/apoptosis. Protein levels of phosphorylated eIF2α, a selective translation inhibitor and UPR hallmark, activating transcription factor 4 (ATF4) and sequestosome-1 were moderately increased, whereas the cell cycle/progression protein cyclin D1 was decreased in HL60. In contrast, EPA concentrations that strongly inhibited and caused activation of the UPR in HL60 cells had no effect on the expression level of these UPR markers in E2R2 cells. Given that the only known difference between these cells is Ec-resistance, our results strongly suggest that the inhibitory effect of EPA on HL60 cells is initially meditated through alterations of the Ca2+-homeostasis followed by activation of the UPR.

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Abbreviations

ATF3, ATF4 and ATF6:

Activating transcription factor 3, 4 and 6

CHOP:

Growth arrest- and DNA damage-inducible gene 153/C/EBP-homologous protein (CHOP/Gadd153)

Ec:

Econazole

EPA:

Eicosapentaenoic acid (20:5n-3)

ER:

Endoplasmic reticulum

ERAD:

ER-associated degradation

eIF2α:

Eukaryote translation initiation factor 2α

EIF2AK3/PERK:

eIF2α kinase 3

HMOX-1:

Heme oxygenase (decycling) 1

IP3:

Inositol 1,4,5-triphosphate

PUFA:

Polyunsaturated fatty acids

SQSTM1:

Sequestosome-1

SOC:

Store-operated Ca2+ channels

UPR:

Unfolded protein response

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Acknowledgments

We thank Professor Stuart A. Berger, University Health Network, Toronto, Canada, for kindly providing the E2R2 cells. Thanks to Gro Leite Størvold, Hilde Bremseth, Anne Gøril Lundemo for technical assistance and Professor Kristian S. Bjerve for good support. The project was financed by The Faculty of Medicine, NTNU, The Cancer Research Fund, Trondheim University Hospital and The Research Council of Norway through grants from the Functional Genomics Program (FUGE). Microarray experiments were performed at the microarray core facility at the Norwegian Microarray Consortium (NMC), Trondheim, which is supported by FUGE, The Norwegian Research Council. Financial support was also given by the cross-disciplinary project “BIOEMIT-Prediction and modification in functional genomics: combining bioinformatical, bioethical, biomedical and biotechnological research”, NTNU.

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Authors and Affiliations

  1. Department of Laboratory Medicine, Children’s and Women’s Health, Norwegian University of Science and Technology (NTNU), Erling Skjalgssons gate 1, 7006, Trondheim, Norway

    Jens Erik Slagsvold, Caroline Hild Hakvåg Pettersen & Svanhild Arentz Schønberg

  2. Department of Mathematical Sciences, Norwegian University of Science and Technology (NTNU), 7491, Trondheim, Norway

    Turid Follestad

  3. Department of Cancer Research and Molecular Medicine, Norwegian University of Science and Technology (NTNU), Erling Skjalgssons gate 1, 7006, Trondheim, Norway

    Hans Einar Krokan

  4. Department of Laboratory Medicine, Children’s and Women’s Health, Norwegian University of Science and Technology, St Olav’s Hospital, 7489, Trondheim, Norway

    Svanhild Arentz Schønberg

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  1. Jens Erik Slagsvold
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Correspondence to Svanhild Arentz Schønberg.

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Slagsvold, J.E., Pettersen, C.H.H., Follestad, T. et al. The Antiproliferative Effect of EPA in HL60 Cells is Mediated by Alterations in Calcium Homeostasis. Lipids 44, 103–113 (2009). https://doi.org/10.1007/s11745-008-3263-5

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  • DOI: https://doi.org/10.1007/s11745-008-3263-5

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