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(Journal of Leukocyte Biology. 2003;73:399-406.)
© 2003 by Society for Leukocyte Biology

Possible contribution of apoptosis-inducing factor (AIF) and reactive oxygen species (ROS) to UVB-induced caspase-independent cell death in the T cell line Jurkat

Hideaki Murahashi^*,, Hiroshi Azuma^*, Naoufal Zamzami, Ko-ji Furuya, Kenji Ikebuchi^*, Miki Yamaguchi^*, Yoshiko Yamada^*, Norihiro Sato^*, Mitsuhiro Fujihara^*, Guido Kroemer and Hisami Ikeda^*

^* Hokkaido Red Cross Blood Center, Sapporo, Japan;
Research and Development Laboratory, Nipro Corporation, Kusatsu, Japan;
Hokkaido Institute of Public Health, Sapporo, Japan; and
CNRS-UMR 1599, Institut Gustave Roussy, Villejuif, France

Correspondence: Hiroshi Azuma, M.D., Hokkaido Red Cross Blood Center, Yamanote 2-2 Nishi-ku, Sapporo 063-0002, Japan. E-mail: azuma{at}hokkaido.bc.jrc.or.jp

We analyzed the mechanism of UVB-induced cell death using the Jurkat T cell line. Apoptosis was assessed by measuring phosphatidylserine (PS) externalization, caspase activity, the decrease in mitochondrial membrane potential (m), nucleosomal DNA fragmentation, and morphological changes such as chromatin condensation. The mitochondrio-nuclear translocation of apoptosis-inducing factor (AIF) was evaluated by confocal laser microscopy. The cell death pattern of UVB-irradiated cells was similar to the Fas-induced cell death pattern. However, zVAD-fmk inhibited the nucleosomal fragmentation of DNA but not the externalization of PS, decrease in m, or mitochondrio-nuclear translocation of AIF. N-acetyl L-cysteine significantly inhibited the translocation of AIF induced by UVB. These results suggested that caspase-dependent and -independent pathways were involved in UVB-induced cell death in Jurkat cells, and the mitochondrio-nuclear translocation of AIF was associated with the latter pathway. In addition, reactive oxygen species generated by UVB might be involved in inducing the mitochondrio-nuclear translocation of AIF.

Key Words: reactive oxygen species • mitochondria • caspase inhibitor • N-acetyl L-cysteine

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