Nanoparticle carbon black driven DNA damage induces growth arrest and AP-1 and NFkappaB DNA binding in lung epithelial A549 cell line.
To assess whether nanoparticle (NP) driven DNA damage induces the expression of proinflammatory transcription factors such as NFB and AP-1 A549, lung epithelial cells were treated with Carbon Black (CB), nanoparticulate CB (NPCB), NPCB coated with BaP (BaP-NPCB) for various times ranging from 30 min to 24 h. DNA strand break was determined by the comet assay and cell cycle status was analyzed using flow cytometry. Nuclear extracts were used for WB analysis of P approximately Ser15-p53. EMSA was used to detect DNA binding. Tested NP caused single strand breaks and significantly altered cell cycle kinetics. NF-kappaB and AP-1 DNA binding were increased at early time points (2.3 and 2.6 fold at 1 hour, respectively). Effects were also found on Ser15-p53 phosphorylation. N-acetylcysteine blocked NP driven effects. In conclusion, NPCB and BaP-NPCB induce DNA damage, activating p53, proteins related to DNA repair and proinflammatory transcription factors.
Mroz
R M
RM
ELEGI/Colt Laboratories, MRC/University of Edinburgh Center for Inflammation Research, Edinburgh, UK. robmroz@wp.pl
Schins
R P F
RP
Li
H
H
Drost
E M
EM
Macnee
W
W
Donaldson
K
K
D000111:Acetylcysteine / Q000494:pharmacology; D002453:Cell Cycle / Q000187:drug effects; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000187:drug effects / Q000648:ultrastructure; D002470:Cell Survival / Q000187:drug effects; D004247:DNA / Q000378:metabolism; D004249:DNA Damage / Q000502:physiology*; D024202:Electrophoretic Mobility Shift Assay; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008168:Lung / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D016328:NF-kappa B / Q000378:metabolism*; D053758:Nanoparticles / Q000633:toxicity*; D010766:Phosphorylation; D053260:Soot / Q000633:toxicity*; D013535:Suspensions; D018808:Transcription Factor AP-1 / Q000378:metabolism*; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism
D016328:NF-kappa B; D053260:Soot; D013535:Suspensions; D018808:Transcription Factor AP-1; D016159:Tumor Suppressor Protein p53; D004247:DNA; D007770:L-Lactate Dehydrogenase; D000111:Acetylcysteine