Regulation of early signaling and gene expression in the alpha-particle and bystander response of IMR-90 human fibroblasts.
BACKGROUND
The existence of a radiation bystander effect, in which non-irradiated cells respond to signals from irradiated cells, is well established. To understand early signaling and gene regulation in bystander cells, we used a bio-informatics approach, measuring global gene expression at 30 minutes and signaling pathways between 30 minutes and 4 hours after exposure to alpha-particles in IMR-90 fibroblasts.


METHODS
We used whole human genome microarrays and real time quantitative PCR to measure and validate gene expression. Microarray analysis was done using BRB-Array Tools; pathway and ontology analyses were done using Ingenuity Pathway Analysis and PANTHER, respectively. We studied signaling in irradiated and bystander cells using immunoblotting and semi-quantitative image analysis.


RESULTS
Gene ontology suggested signal transduction and transcriptional regulation responding 30 minutes after treatment affected cell structure, motility and adhesion, and interleukin synthesis. We measured time-dependent expression of genes controlled by the NF-kappaB pathway; matrix metalloproteinases 1 and 3; chemokine ligands 2, 3 and 5 and interleukins 1beta, 6 and 33. There was an increased response of this set of genes 30 minutes after treatment and another wave of induction at 4 hours. We investigated AKT-GSK3beta signaling and found both AKT and GSK3beta are hyper-phosphorylated 30 minutes after irradiation and this effect is maintained through 4 hours. In bystander cells, a similar response was seen with a delay of 30 minutes. We proposed a network model where the observed decrease in phosphorylation of beta-catenin protein after GSK3beta dependent inactivation can trigger target gene expression at later times after radiation exposure


CONCLUSIONS
These results are the first to show that the radiation induced bystander signal induces a widespread gene expression response at 30 minutes after treatment and these changes are accompanied by modification of signaling proteins in the PI3K-AKT-GSK3beta pathway.
Ghandhi
Shanaz A
SA
Center for Radiological Research, Columbia University, VC11-215, 630 West 168th Street, New York, NY 10032, USA. saa2108@columbia.edu
Ming
Lihua
L
Ivanov
Vladimir N
VN
Hei
Tom K
TK
Amundson
Sally A
SA
D000512:Alpha Particles; D024201:Bystander Effect; D002460:Cell Line; D005347:Fibroblasts / Q000378:metabolism* / Q000528:radiation effects; D005786:Gene Expression Regulation / Q000528:radiation effects*; D053263:Gene Regulatory Networks; D038362:Glycogen Synthase Kinase 3 / Q000235:genetics / Q000378:metabolism; D000071679:Glycogen Synthase Kinase 3 beta; D006801:Humans; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction / Q000528:radiation effects*; D013997:Time Factors; D051176:beta Catenin / Q000235:genetics / Q000378:metabolism
D016328:NF-kappa B; D051176:beta Catenin; C000605506:GSK3B protein, human; D000071679:Glycogen Synthase Kinase 3 beta; D038362:Glycogen Synthase Kinase 3