Cell Biology and Toxicology

, Volume 26, Issue 4, pp 331–339

Morphological damages of a glyphosate-treated human keratinocyte cell line revealed by a micro- to nanoscale microscopic investigation

  • Celine Elie-Caille
  • Celine Heu
  • Catherine Guyon
  • Laurence Nicod
Article

DOI: 10.1007/s10565-009-9146-6

Cite this article as:
Elie-Caille, C., Heu, C., Guyon, C. et al. Cell Biol Toxicol (2010) 26: 331. doi:10.1007/s10565-009-9146-6

Abstract

Among the molecules to which the human skin is exposed, glyphosate is used as an herbicide. Glyphosate has been shown to induce in vitro cutaneous cytotoxic effects, concomitant with oxidative disorders. In this following study, we focused on dynamic events of the loss of HaCaT cell integrity appearing after a glyphosate treatment. In these conditions, we showed that glyphosate is able to disrupt HaCaT cells and to induce intracellular oxidative cascade. In this aim, we optimized the conditions of cell treatment playing on exposure time (from 24 h to 30 min), which directly modify the cell viability profile (glyphosate 50% inhibition concentration from 28 to 53 mM) and allow to track cells along the treatment as an “induction and visualization” process. The combination of atomic force and fluorescence microscopic approaches offered opportunities to lead in parallel an investigation of the membrane surface and of the intracellular disorders, through cytoskeleton, nuclear, and oxidative stress marker targeting. The originality of our approach relies on monitoring all events derived from oxidative stress in process and performed by simultaneous cytotoxic induction and nanoscale cell visualization. We revealed a transition from spread and globular to elongated cell morphology, with a drastic cell size reduction, after a dose- and time-dependent glyphosate treatment; a redistribution of cell surface protrusions was also pointed out. All these membrane damages, added to observations of disorganized cytoskeleton, condensed chromatin, and overproduction of oxidative reactive species, lead us to conclude that glyphosate acts in induction of apoptotic process.

Keywords

HaCaTGlyphosateCytotoxicityCell integrityApoptosisConfocal and atomic force microscopy

Abbreviations

AFM

Atomic force microscopy

DCFH-DA

2′,7′-Dichlorodihydrofluorescein diacetate

DMEM

Dulbecco’s modified Eagle’s medium

FCS

Fetal calf serum

FITC

Fluorescein isothiocyanate

IC50

50% inhibition concentration

MTT

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

PBS

Phosphate-buffered saline

PFA

p-Formaldehyde

UVB

Ultraviolet B

Copyright information

© Springer Science+Business Media B.V. 2009

Authors and Affiliations

  • Celine Elie-Caille
    • 1
  • Celine Heu
    • 1
    • 2
  • Catherine Guyon
    • 2
  • Laurence Nicod
    • 2
  1. 1.Clinical & Innovation Proteomic Platform (CLIPP), Institut Femto-st, UMR 6174 CNRSUniversity of Franche-ComteBesançon cedexFrance
  2. 2.Laboratoire de Biologie Cellulaire, EA4267, IFR133, UFR Sciences Médicales & Pharmaceutiques, Place St-JacquesUniversity of Franche-ComteBesançon cedexFrance