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Biological Trace Element Research

, Volume 154, Issue 2, pp 288–298 | Cite as

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

  • Florence Hazane-PuchEmail author
  • Pierre Champelovier
  • Josiane Arnaud
  • Catherine Garrel
  • Bruno Ballester
  • Patrice Faure
  • François Laporte
Article

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.

Keywords

Selenomethionine Sodium selenite HaCaT cells Antioxidant UVA Cell death 

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

Notes

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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Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Florence Hazane-Puch
    • 1
    Email author
  • Pierre Champelovier
    • 2
  • Josiane Arnaud
    • 1
  • Catherine Garrel
    • 1
  • Bruno Ballester
    • 3
  • Patrice Faure
    • 1
    • 4
  • François Laporte
    • 1
  1. 1.Unité de Biochimie Hormonale et Nutritionnelle, Département de Biochimie, Toxicologie et Pharmacologie, Institut de Biologie et de PathologieCentre Hospitalier Universitaire de GrenobleGrenobleFrance
  2. 2.Unité de Cytologie, Département d’Anatomie et de Cytologie Pathologiques, Institut de Biologie et de PathologieCentre Hospitalier Universitaire de GrenobleGrenobleFrance
  3. 3.CRSSA/IRBA, antenne La TroncheLa Tronche CedexFrance
  4. 4.Laboratoire d’étude de la physiopathologie de l’hypoxie (HP2), Inserm U 1042, Faculté de médecine et de pharmacieLa TroncheFrance

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