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Sphingosine 1‐phosphate attenuates peroxide‐induced apoptosis in HaCaT cells cultured in vitro
Author(s) -
Moriue T.,
Igarashi J.,
Yoneda K.,
Hashimoto T.,
Nakai K.,
Kosaka H.,
Kubota Y.
Publication year - 2013
Publication title -
clinical and experimental dermatology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.587
H-Index - 78
eISSN - 1365-2230
pISSN - 0307-6938
DOI - 10.1111/ced.12037
Subject(s) - hacat , wortmannin , apoptosis , protein kinase b , microbiology and biotechnology , sphingosine 1 phosphate , programmed cell death , pi3k/akt/mtor pathway , sphingosine , lipid signaling , chemistry , tunel assay , biology , signal transduction , biochemistry , in vitro , receptor
Summary Background Sphingosine 1‐phosphate (S1P) is a sphingolipid mediator that elicits a wide array of physiological responses in various types of mammalian cells. Among the numerous biological activities elicited by S1P is protection from apoptotic cell death, which seems to take place through the cell‐surface S1P receptor and the downstream phosphoinositide 3′‐ OH kinase ( PI 3‐K)/Akt pathway. It is unclear whether and how S1P protects human keratinocytes from hydrogen peroxide (H 2 O 2 )‐induced apoptosis. Aim We investigated the effects of S1P on apoptotic cell death in HaCaT cells, spontaneously immortalized human keratinocytes. Methods HaCaT cells were treated with hydrogen peroxide (H 2 O 2 ) 1–2 mmol/L as an inducer of apoptosis. Cellular apoptosis was assessed with terminal dUTP nick‐end labelling (TUNEL), WST‐8 and immunoblot assays. Results In WST ‐8 and TUNEL assays, S1P pretreatment (1 μmol/L for 30 min) attenuated H 2 O 2 ‐induced cell death. Promotion of the cleavage of caspase‐3 by H 2 O 2 was markedly attenuated when cells had been preincubated with S1P. S1P markedly potentiated phosphorylation (activation) of Akt in the presence of H 2 O 2 . Wortmannin, a selective inhibitor of the PI 3‐K/Akt pathway, significantly suppressed S1P‐induced attenuation of caspase‐3 cleavage promoted by H 2 O 2 . Conclusions S1P, a sphingolipid mediator, attenuates H 2 O 2 ‐induced apoptosis of HaCaT cells, by promoting phosphorylation of the Akt pathway.