title: Acute activation of AMP-activated protein kinase prevents H2O2-induced premature senescence in primary human keratinocytes.
abstract: We investigated the effects of AMPK on H(2)O(2)-induced premature senescence in primary human keratinocytes. Incubation with 50 µM H(2)O(2) for 2 h resulted in premature senescence with characteristic increases in senescence-associated ß-galactosidase (SA-gal) staining 3 days later and no changes in AMPK or p38 MAPK activity. The increase in SA-gal staining was preceded by increases in both p53 phosphorylation (S15) (1 h) and transactivation (6 h) and the abundance of the cyclin inhibitor p21(CIP1) (16 h). Incubation with AICAR or resveratrol, both of which activated AMPK, prevented the H(2)O(2)-induced increases in both SA-Gal staining and p21 abundance. In addition, AICAR diminished the increase in p53 transactivation. The decreases in SA-Gal expression induced by resveratrol and AICAR were prevented by the pharmacological AMPK inhibitor Compound C, expression of a DN-AMPK or AMPK knock-down with shRNA. Likewise, both knockdown of AMPK and expression of DN-AMPK were sufficient to induce senescence, even in the absence of exogenous H(2)O(2). As reported by others, we found that AMPK activation by itself increased p53 phosphorylation at S15 in embryonic fibroblasts (MEF), whereas under the same conditions it decreased p53 phosphorylation in the keratinocytes, human aortic endothelial cells, and human HT1080 fibrosarcoma cells. In conclusion, the results indicate that H(2)O(2) at low concentrations causes premature senescence in human keratinocytes by activating p53-p21(CIP1) signaling and that these effects can be prevented by acute AMPK activation and enhanced by AMPK downregulation. They also suggest that this action of AMPK may be cell or context-specific.
authors.affiliation: Section of Endocrinology and Diabetes Research Unit, Department of Medicine, Boston University School of Medicine, Boston, Massachusetts, United States of America. yido@bu.edu
authors.forename: Yasuo
authors.initials: Y
authors.lastname: Ido
authors.forename: Albert
authors.initials: A
authors.lastname: Duranton
authors.forename: Fan
authors.initials: F
authors.lastname: Lan
authors.forename: Jose M
authors.initials: JM
authors.lastname: Cacicedo
authors.forename: Tai C
authors.initials: TC
authors.lastname: Chen
authors.forename: Lionel
authors.initials: L
authors.lastname: Breton
authors.forename: Neil B
authors.initials: NB
authors.lastname: Ruderman
mesh_terms: D055372:AMP-Activated Protein Kinases / Q000235:genetics / Q000378:metabolism*; D000256:Adenoviridae / Q000235:genetics; D000620:Aminoimidazole Carboxamide / Q000031:analogs & derivatives / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects; D016922:Cellular Senescence / Q000187:drug effects*; D004789:Enzyme Activation / Q000187:drug effects; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D015603:Keratinocytes / Q000187:drug effects* / Q000378:metabolism*; D012265:Ribonucleotides / Q000494:pharmacology; D015398:Signal Transduction / Q000187:drug effects
chemical_list: D012265:Ribonucleotides; D000620:Aminoimidazole Carboxamide; D006861:Hydrogen Peroxide; D055372:AMP-Activated Protein Kinases; C031143:AICA ribonucleotide