Initiation of rapid, P53-dependent growth arrest in cultured human skin fibroblasts by reactive chlorine species.
Oxidants produced by neutrophils have been implicated in causing cancers associated with chronic inflammation. Hypochlorous acid is the most potent oxidant produced by these cells in appreciable amounts. It reacts with amines to form chloramines, which are weaker oxidants but are mutagenic. Recently, we showed that sublethal doses of hypochlorous acid increased levels of the transcription factor protein 53 (p53) and the wild-type activating fragment-1/cyclin-dependent kinase inhibitory protein-1 (WAF1/CIP1) in cultured human skin fibroblasts. WAF1/CIP1 is an important intermediate in the pathway leading to growth arrest. We now show that low doses of hypochlorous acid and physiological chloramines lead to an inhibition of both DNA synthesis and division of cultured human skin fibroblasts. Inhibition of DNA synthesis occurred within 1 h of hypochlorous acid treatment, was maintained for 24 h, and returned to a normal rate after 48 h. Cell division was inhibited by hypochlorous acid and chloramines for 48 h and returned to normal 72 h after treatment. Growth arrest was dependent on p53 because it was blocked when cells were transfected with a p53-binding oligonucleotide. We propose that reactive chlorine species will initiate WAF1/CIP1-dependent growth arrest that will counteract their mutagenic effects and minimize the possibility of the malignant transformation of cells surrounding sites of inflammation.
Vile
G F
GF
Free Radical Research Group, Christchurch School of Medicine, Christchurch, New Zealand.
Rothwell
L A
LA
Kettle
A J
AJ
D002455:Cell Division / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002700:Chloramines / Q000008:administration & dosage / Q000633:toxicity*; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins / Q000378:metabolism; D004247:DNA / Q000096:biosynthesis; D004305:Dose-Response Relationship, Drug; D005347:Fibroblasts / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D005998:Glycine / Q000008:administration & dosage / Q000031:analogs & derivatives / Q000633:toxicity; D006801:Humans; D006997:Hypochlorous Acid / Q000008:administration & dosage / Q000633:toxicity*; D007249:Inflammation / Q000378:metabolism; D018836:Inflammation Mediators / Q000008:administration & dosage / Q000633:toxicity; D012867:Skin; D013654:Taurine / Q000008:administration & dosage / Q000031:analogs & derivatives / Q000633:toxicity; D015533:Transcriptional Activation; D014162:Transfection; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism*
C493956:CDKN1A protein, human; D002700:Chloramines; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins; D018836:Inflammation Mediators; D016159:Tumor Suppressor Protein p53; D013654:Taurine; C012826:glycine chloramine; C043410:N-chlorotaurine; D006997:Hypochlorous Acid; D004247:DNA; C030816:chloramine; D005998:Glycine