Membrane-dependent bystander effect contributes to amplification of the response to alpha-particle irradiation in targeted and nontargeted cells.
OBJECTIVE
Free radicals are believed to play an active role in the bystander response. This study investigated their origin as well as their temporal and spatial impacts in the bystander effect.


METHODS
We employed a precise alpha-particle microbeam to target a small fraction of subconfluent osteoblastic cells (MC3T3-E1). gammaH2AX-53BP1 foci, oxidative metabolism changes, and micronuclei induction in targeted and bystander cells were assessed.


RESULTS
Cellular membranes and mitochondria were identified as two distinct reactive oxygen species producers. The global oxidative stress observed after irradiation was significantly attenuated after cells were treated with filipin, evidence for the primal role of membrane in the bystander effect. To determine the membrane's impact at a cellular level, micronuclei yield was measured when various fractions of the cell population were individually targeted while the dose per cell remained constant. Induction of micronuclei increased in bystander cells as well as in targeted cells and was attenuated by filipin treatment, demonstrating a role for bystander signals between irradiated cells in an autocrine/paracrine manner.


CONCLUSIONS
A complex interaction of direct irradiation and bystander signals leads to a membrane-dependent amplification of cell responses that could influence therapeutic outcomes in tissues exposed to low doses or to environmental exposure.
CEA, IRAMIS, SIS2M, F-91191 Gif-sur-Yvette, France. maite.hanot@cea.fr
Maïté
M
Hanot
Jim
J
Hoarau
Marie
M
Carrière
Jaime F
JF
Angulo
Hicham
H
Khodja
D016475:3T3 Cells; D000512:Alpha Particles / Q000627:therapeutic use; D000818:Animals; D024201:Bystander Effect / Q000187:drug effects / Q000502:physiology*; D002462:Cell Membrane / Q000378:metabolism*; D053903:DNA Breaks, Double-Stranded; D005372:Filipin / Q000494:pharmacology; D005978:Glutathione / Q000378:metabolism; D006657:Histones / Q000378:metabolism; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism; D051379:Mice; D015162:Micronucleus Tests; D008928:Mitochondria / Q000378:metabolism*; D010006:Osteoblasts / Q000378:metabolism / Q000528:radiation effects / Q000648:ultrastructure; D018384:Oxidative Stress / Q000187:drug effects; D026361:Reactive Nitrogen Species / Q000378:metabolism*; D017382:Reactive Oxygen Species / Q000378:metabolism*; D000071857:Tumor Suppressor p53-Binding Protein 1
C468783:H2AX protein, human; D006657:Histones; D047908:Intracellular Signaling Peptides and Proteins; D026361:Reactive Nitrogen Species; D017382:Reactive Oxygen Species; C498301:TP53BP1 protein, human; D000071857:Tumor Suppressor p53-Binding Protein 1; D005372:Filipin; D005978:Glutathione