H(2)O(2)-induced AP-1 activation and its effect on p21(WAF1/CIP1)-mediated G2/M arrest in a p53-deficient human lung cancer cell.

Abstract

Cellular response to oxidative stress is a complex process that is often connected to cell cycle regulation. The present study examines the effect of H(2)O(2) on cell cycle regulation and involvement of reactive oxygen species (ROS) in these H(2)O(2)-induced responses in a p53-deficient human lung carcinoma cell line, H1299. Treatment of the cells with H(2)O(2) caused a G2/M phase arrest. Among the redox-sensitive transcription factors, NF-kappaB and AP-1, we found that only AP-1 was activated by 200 microM H(2)O(2) in human lung cells. Furthermore, electrophoretic mobility shift assays revealed that H(2)O(2) enhanced the DNA binding of AP-1 to a putative AP-1 binding element (TGAGGAA) in the p21(WAF1/CIP1) promoter region (between -2203 and -2197 nucleotides upstream of the transcription initiation site). An increase in c-Jun phosphorylation by ERK was also found to accompany the increased AP-1 activity as detected by Western blot. PD98059, a specific inhibitor of MEK, diminished H(2)O(2)-induced phosphorylation of c-Jun and DNA binding activity of AP-1, decreased expression of p21(WAF1/CIP1), and released the cells from G2/M arrest. Taken together, these results revealed a novel AP-1 binding site in the promoter region of p21(WAF1/CIP1) and a possible cell cycle regulation mechanism mediated by activation of a redox-dependent ERK signaling pathway.

• Authors: Youn Wook Chung, Dae-won Jeong, Joo Yun Won, Eui-Ju Choi, Yung Hyun Choi, Ick Young Kim
• Journal: Biochemical and biophysical research communications (Biochem Biophys Res Commun) Vol. 293 Issue 4 Pg. 1248-53 (May 17 2002) ISSN: 0006-291X [Print] United States
• PMID: 12054510 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: (c) 2002 Elsevier Science (USA).

Chemical References

• CDKN1A protein, human
• Cyclin-Dependent Kinase Inhibitor p21
• Cyclins
• Enzyme Inhibitors
• Flavonoids
• Proto-Oncogene Proteins c-jun
• Reactive Oxygen Species
• Transcription Factor AP-1
• Hydrogen Peroxide
• Luciferases
• Mitogen-Activated Protein Kinases
• 2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one

Topics

• Binding Sites
• Blotting, Western
• Cell Cycle
• Cell Nucleus (metabolism)
• Cell Separation
• Cyclin-Dependent Kinase Inhibitor p21
• Cyclins (genetics, metabolism)
• Enzyme Inhibitors (pharmacology)
• Flavonoids (pharmacology)
• Flow Cytometry
• G2 Phase (drug effects)
• Genes, p53
• Humans
• Hydrogen Peroxide (pharmacology)
• Luciferases (metabolism)
• Lung Neoplasms (drug therapy, genetics)
• Mitogen-Activated Protein Kinases (metabolism)
• Mitosis (drug effects)
• Phosphorylation
• Promoter Regions, Genetic
• Protein Binding
• Proto-Oncogene Proteins c-jun (metabolism)
• Reactive Oxygen Species
• Signal Transduction
• Transcription Factor AP-1 (metabolism)
• Tumor Cells, Cultured