title: Akt1-mediated intracellular oxidation after UVB irradiation suppresses apoptotic cell death induced by cell detachment and serum starvation.
abstract: Apoptosis is an important cell death system that deletes damaged and mutated cells to prevent cancer. We have previously reported that a certain dose of UVB irradiation inhibited the apoptosis induced by serum starvation and cell detachment, leading to cell transformation. This antiapoptotic effect was partially inhibited by phosphatidylinositol 3-kinase (PI3-kinase) inhibitors. UVB irradiation is known to cause the phosphorylation of Akt via the activation of PI3-kinase; however, the Akt isoform-specific relationship has not yet been clarified. Notably, the role in antiapoptotic effect of UVB has yet to be elucidated. In this study, the role of Akt1 in the UVB-induced inhibition of apoptosis was examined by Akt1 knockdown using small interfering RNA (siRNA). NIH3T3 cells showed typical apoptotic cell death by serum starvation and cell detachment, which was significantly inhibited by UVB irradiation. Akt1 knockdown decreased the antiapoptotic effect of UVB. Hydrogen peroxide-induced suppression of cell death was also decreased in Akt1 knockdown cells. An antioxidant, N-acetylcysteine, inhibited the antiapoptotic effect by UVB irradiation, whereas no inhibition was observed in Akt1 knockdown cells. Furthermore, UVB-induced intracellular peroxidation was not observed in the knockdown cells, indicating that Akt1 played an important role in mediating the intracellular redox status. Treatment with insulin had a similar antiapoptotic effect as UVB irradiation involving intracellular peroxidation, which was also attenuated in Akt1 knockdown cells. These findings suggest that appropriate intracellular oxidation after UVB irradiation prevented apoptosis, a process which might be partially regulated by the production of reactive oxygen species mediated by Akt1.
authors.affiliation: Laboratory of Radiation Biology, Graduate School of Nutritional and Environmental Sciences, University of Shizuoka, Shizuoka-shi, Japan. ibuki@u-shizuoka-ken.ac.jp
authors.forename: Yuko
authors.initials: Y
authors.lastname: Ibuki
authors.forename: Marie
authors.initials: M
authors.lastname: Akaike
authors.forename: Tatsushi
authors.initials: T
authors.lastname: Toyooka
authors.forename: Rensuke
authors.initials: R
authors.lastname: Goto
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000528:radiation effects*; D020169:Caspases / Q000378:metabolism; D002448:Cell Adhesion / Q000528:radiation effects; D016895:Culture Media, Serum-Free; D004789:Enzyme Activation; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects / Q000235:genetics; D051379:Mice; D041681:NIH 3T3 Cells; D010084:Oxidation-Reduction / Q000528:radiation effects; D051057:Proto-Oncogene Proteins c-akt / Q000235:genetics / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000235:genetics; D017382:Reactive Oxygen Species / Q000378:metabolism
chemical_list: D016895:Culture Media, Serum-Free; D034741:RNA, Small Interfering; D017382:Reactive Oxygen Species; D051057:Proto-Oncogene Proteins c-akt; D020169:Caspases