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Long-Term Selenium Supplementation in HaCaT Cells: Importance of Chemical Form for Antagonist (Protective Versus Toxic) Activities

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Abstract

The beneficial effect of selenium (Se) on cancer is known to depend on the chemical form, the dose and the duration of the supplementation. The aim of this work was to explore long term antagonist (antioxidant versus toxic) effects of an inorganic (sodium selenite, Na2SeO3) and an organic (seleno-L-methionine, SeMet) forms in human immortalized keratinocytes HaCaT cells. HaCaT cells were supplemented with Na2SeO3 or SeMet at micromolar concentrations for 144 h, followed or not by UVA radiation. Se absorption, effects of UVA radiation, cell morphology, antioxidant profile, cell cycle processing, DNA fragmentation, cell death triggered and caspase-3 activity were determined. At non-toxic doses (10 μM SeMet and 1 μM Na2SeO3), SeMet was better absorbed than Na2SeO3. The protection of HaCaT from UVA-induced cell death was observed only with SeMet despite both forms increased glutathione peroxidase-1 (GPX1) activities and selenoprotein-1 (SEPW1) transcript expression. After UVA irradiation, malondialdehyde (MDA) and SH groups were not modulated whatever Se chemical form. At toxic doses (100 μM SeMet and 5 μM Na2SeO3), Na2SeO3 and SeMet inhibited cell proliferation associated with S-G2 blockage and DNA fragmentation leading to apoptosis caspase-3 dependant. SeMet only led to hydrogen peroxide production and to a decrease in mitochondrial transmembrane potential. Our study of the effects of selenium on HaCaT cells reaffirm the necessity to take into account the chemical form in experimental and intervention studies.

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Abbreviations

DMSO:

dimethylsulfoxide

CDK:

cycline dependant kinase

DCFDA:

2′,7′-dichlorofluorescein diacetate

DiOC6:

3,3′-dihexyloxacarbocyanine iodide

FCS:

foetal calf serum

GADD:

growth arrest and DNA damage

GPX1:

glutathione peroxidase 1

GSR:

glutathione reductase

H2Se:

selenide

HBSS:

Hank’s buffered salt solution

HPLC:

high performance liquid chromatography

HPRT1:

hypoxanthine phosphoribosyltransferase 1

ICP-MS:

inductively coupled plasma mass-spectrometry

mTOR:

mammalian target of rapamycin

MDA:

malondialdehyde

MGG:

May–Grünwald Giemsa

MI:

mitotic index

MTP:

mitochondrial transmembrane potential

MTT:

3-(4,5-dimethylthiazol- 2yl)-2,5-diphenyl tetrazolium bromide

NHSF:

normal human skin fibroblast

ROS:

reactive oxygen species

SB:

strand break

SeCys:

selenocysteine

SeMet:

selenomethionine

Na2SeO3 :

sodium selenite

SEPW1:

selenoprotein W1

SH:

thiol groups

SOD:

superoxide dismutase

SP:

selenoprotein

UVA:

ultraviolet-A

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Acknowledgments

We would like to thank all the technicians for their technical assistance: Aurélie Dariz and Michelle Tripier-Champ for cell cultures, toxicity assays and RT-q-PCR, Sandra Grange and Angèle Kraviec for GPX1 activities and SH-groups determination, Laurence Puillet for MDA measurements, Marie-Christine Bouillet and Dominique André for selenium measurements.

Declaration of Interest

The authors declare that there are no conflicts of interest.

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Authors and Affiliations

  1. Unité de Biochimie Hormonale et Nutritionnelle, Département de Biochimie, Toxicologie et Pharmacologie, Institut de Biologie et de Pathologie, Centre Hospitalier Universitaire de Grenoble, CS10217, 38043, Grenoble, France

    Florence Hazane-Puch, Josiane Arnaud, Catherine Garrel, Patrice Faure & François Laporte

  2. Unité de Cytologie, Département d’Anatomie et de Cytologie Pathologiques, Institut de Biologie et de Pathologie, Centre Hospitalier Universitaire de Grenoble, CS10217, 38043, Grenoble, France

    Pierre Champelovier

  3. CRSSA/IRBA, antenne La Tronche, BP87, 38702, La Tronche Cedex, France

    Bruno Ballester

  4. Laboratoire d’étude de la physiopathologie de l’hypoxie (HP2), Inserm U 1042, Faculté de médecine et de pharmacie, Domaine de la Merci, 38700, La Tronche, France

    Patrice Faure

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  1. Florence Hazane-Puch
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Correspondence to Florence Hazane-Puch.

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Hazane-Puch, F., Champelovier, P., Arnaud, J. et al. Long-Term Selenium Supplementation in HaCaT Cells: Importance of Chemical Form for Antagonist (Protective Versus Toxic) Activities. Biol Trace Elem Res 154, 288–298 (2013). https://doi.org/10.1007/s12011-013-9709-5

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